phenanthrenes and Reperfusion-Injury

Cerebral ischemia is a major health threat to humankind around the world, and the reperfusion methods may provoke irreversible damages to brain tissues, causing impairment of neurological function. The goal of this study is to investigate the potential neurological protective effect of PJ34, a well-characterized poly (ADP-ribose) polymerase 1 (PARP-1) inhibitor, on cerebral ischemia-reperfusion (I/R)-induced injury of the rat model. The cerebral I/R rats were received (3, 6, or 12 mg/kg) injections of PJ34 or saline at 24 h, 6 h before middle cerebral artery occlusion (MCAO) and 1 h, 24 h, and 48 h after MCAO. All rats were subject to the neurological behavior tests by open field test and Morris water maze test. The expression of pro-inflammatory cytokines, Cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS) in cerebral tissues was also determined. Our results demonstrated that the administration of PJ34 dose-dependently ameliorated cerebral I/R-induced injury and improved neurological performance of cerebral I/R rats. We also revealed that PJ34 treatment effectively reduced COX2, iNOS, and pro-inflammatory cytokine levels in the I/R-induced injury tissues. Our finding further supports that inhibition of PARP-1 activity is beneficial for reducing post-I/R-induced brain damage via targeting inflammatory response.

Topics: Animals; Brain; Cognitive Dysfunction; Disease Models, Animal; Dose-Response Relationship, Drug; Humans; Inflammation Mediators; Ischemic Attack, Transient; Male; Morris Water Maze Test; Phenanthrenes; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Rats; Reperfusion Injury

The diterpenoid cryptotanshinone (CTS) has wide biological functions, including inhibition of tumor growth, inflammation and apoptosis. The present study aimed to explore the possible effect of CTS on cerebral ischemia/reperfusion (I/R) injury and the underlying mechanisms.. Male C57BL/6J mice underwent transient middle cerebral artery occlusion (tMCAO) and murine microglia BV2 cells were challenged by Oxygen/glucose deprivation, to mimic I/R and ischemic/hypoxic and reperfusion (H/R) injury, respectively. CTS was administered 0.5 h (10 mg/kg) after the onset of MCAO or 2 h (20μM) post OGD. Infarct volume and neurological deficit were measured. Immunofluorescence, qPCR, and western blot, were performed to detect the expression of cytokines, apoptotic marker, and M1/M2 phenotype-specific genes. Flow cytometry was applied for M1/M2 subpopulation or Annexin V/PI apoptosis assessment.. CTS significantly reduced cerebral infarct volume, neurologic deficit scores, pro-inflammatory cytokine production (IL-6, TNF-α, and IL-1β), apoptotic protein expression (cleaved caspase-3) of mice after tMCAO challenge. Furthermore, CTS attenuated CD16+ M1-type and elevated CD206+ M2-type microglia in vivo or in vitro.. We propose that the neuroprotective effect of CTS in the I/R or H/R context are explained modulation of microglial polarization, suggesting therapeutic potential for cerebral ischemic stroke.

Topics: Animals; Infarction, Middle Cerebral Artery; Male; Mice; Mice, Inbred C57BL; Microglia; Phenanthrenes; Reperfusion Injury

Sodium tanshinone IIA sulfonate (STS) can protect against brain damage induced by stroke. However, the neural protection mechanism of STS remains unclear. We investigated whether STS performs its protective function by suppressing autophagy and inflammatory activity during brain injury. We established a transient middle cerebral artery occlusion and reperfusion (MCAO/R) model by blocking the left middle cerebral artery with a thread inserted through the internal carotid artery for 1 h, followed by reperfusion for 48 h either with or without STS and the autophagy inhibitor 3-methyladenine (3-MA). Neuroprotective effects were determined by evaluating infarction, brain edema, and neurological deficits. The numbers of microglia-derived macrophages, monocyte-derived microglia, T cells, and B cells in the brains were measured, based on the surface marker analyses of CD45, CD11b, B220, CD3, and CD4 using fluorescence-assisted cell sorting. STS (10, 20, 40 mg/kg) was able to significantly reduce infarct volumes, improve neurological deficits, and reduce brain water contents. STS treatment reduced neuroinflammation, as assessed by the infiltration of macrophages and neutrophils, corresponding with reduced numbers of macrophages, T cells, and B cells in ischemia/reperfusion (I/R) brains. In addition, STS treatment also attenuated the upregulation of autophagy associated proteins, such as LC3-II, Beclin-1 and Sirt 6, which was induced by MCAO. These results demonstrated that STS can provide remarkable protection against ischemic stroke, possibly via the inhibition of autophagy and inflammatory activity.

Topics: Animals; Autophagy; Brain Ischemia; Disease Models, Animal; Infarction, Middle Cerebral Artery; Inflammation; Neuroprotective Agents; Phenanthrenes; Rats; Rats, Sprague-Dawley; Reperfusion Injury

Triptolide, extracted from Chinese medicinal materials Tripterygium wilfordii Hook F (TwHF), has immunosuppressive, anti-inflammatory and anti-tumour effects. The purpose of this study was to examine whether triptolide has the neuroprotective effect on cerebral ischemia-reperfusion (I/R) injury and to explore its possible mechanism.. The rat model of focal cerebral I/R was established by the suture-occluded method. The SD rats were randomly divided into five groups: sham operation group (Sham group), ischemia-reperfusion model group (I/R group), low concentration of triptolide group (12.5 mg/kg, TL-L group), medium concentration of triptolide group (25 mg/kg, TL-M group) and high concentration of triptolide group (50 mg/kg, TL-H group). The neurological function of the rats was scored, the degree of brain oedema was detected by the dry-wet method, and the cerebral infarction area was determined by TTC staining. Nissl staining was used to detect neuronal damage. The contents of reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD) were also detected. Meanwhile, the expression level of proteins related to Wnt/-catenin signalling pathway was measured by Western blot.. Compared with the I/R group, cerebral oedema, infarct volume, neurological impairment, the contents of MDA and ROS were reduced, while the SOD level was increased in the TL-L, TL-M, and TL-H groups. The results of Nissl staining showed that triptolide could reduce the nerve cell injury caused by cerebral I/R. In addition, the results of Western blot confirmed that the expression of Wnt1, -catenin, c-Myc, and Cyclin-D1 were down-regulated after triptolide intervention, that is, inhibited the activation of Wnt/-catenin signalling pathway.. Triptolide mediates Wnt/-catenin signalling pathway to alleviate cerebral I/R injury in rats. This study provides ideas and experimental basis for the treatment of ischemic stroke patients.

Topics: Animals; Brain; Brain Ischemia; Diterpenes; Epoxy Compounds; Immunosuppressive Agents; Male; Neuroprotective Agents; Phenanthrenes; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Wnt Signaling Pathway

Cryptotanshinone (CTS), an active component extracted from the root of Salvia miltiorrhiza Bunge , was reported to attenuate hepatic ischemia/reperfusion (I/R) injury. However, its protective effect against renal I/R injury remains unclear. In this study, the role of CTS in renal I/R injury in vitro and its possible mechanism were investigated. Our results showed that CTS improved cell viability in HK-2 cells exposed to hypoxia/reoxygenation (H/R). CTS also inhibited the H/R-mediated production of reactive oxygen species, as well as increased the activities of superoxide dismutase and catalase in H/R-stimulated HK-2 cells. In addition, CTS dramatically attenuated the induction of bax expression and caspase-3 activity and alleviated the reduction of bcl-2 expression in HK-2 cells cultured with H/R. Furthermore, CTS activated the levels of p-PI3K and p-Akt in H/R-injured HK-2 cells; meanwhile, the renal protective activity of CTS was inhibited by the inhibitor of the (phosphatidylinositol 3 kinase/protein kinase B) PI3K/Akt pathway (LY294002). These findings indicate that CTS can ameliorate renal I/R injury in vitro partly through regulating the PI3K/Akt pathway.

Topics: Apoptosis; Cell Line; Cell Survival; Humans; Oxidative Stress; Phenanthrenes; Reactive Oxygen Species; Reperfusion Injury; Signal Transduction

Cryptotanshinone (CTS) is a natural compound from the Chinese herb Salvia miltiorrhiza. Previous studies demonstrated that CTS possesses anti-apoptotic and anti-inflammatory properties. However, its effects and underlying mechanism on renal ischaemia reperfusion (IR) injury remain unknown. In the present study, we investigated the effects of CTS on renal IR injury and its potential underlying mechanisms. Mice were randomized into four groups as follows: (a) sham operation + vehicle, (b) sham operation + CTS, (c) IR + vehicle, (d) IR + CTS. The CTS-treated group were injected intraperitoneally with CTS (10 mg/kg/d) for 7 days prior to IR operation. Renal IR injury was induced by clamping the bilateral renal artery for 30 minutes followed by 24 hours of reperfusion. The mice were then killed to collect the serum and the kidneys for analysis. The results of the present study showed that CTS pretreatment significantly attenuates IR-induced renal functional and morphological injuries, which was accompanied with inhibition of cell apoptosis and inflammatory response. Moreover, the phosphorylation of p38 mitogen-activated protein kinase (MAPK) and the activation of nuclear factor-κB (NF-κB) signalling were inhibited by CTS. Therefore, CTS could be a useful therapeutic agent in the fight against renal IR injury.

Topics: Animals; Anti-Inflammatory Agents; Apoptosis; Disease Models, Animal; Humans; Inflammation; Injections, Intraperitoneal; Kidney; Male; Mice; p38 Mitogen-Activated Protein Kinases; Phenanthrenes; Phosphorylation; Reperfusion Injury; Signal Transduction; Tumor Necrosis Factor-alpha

Intestinal dysfunction, especially acute pathologies linked to intestinal ischemia/reperfusion (I/R) injury, is profoundly affected by inflammation and improper execution of cell death. Few studies have examined the efficacy of combined strategies in regulated intestinal epithelial necrosis after intestinal I/R. Here, we evaluated the functional interaction between poly (adenosine diphosphate-ribose) polymerase 1 (PARP-1)-induced parthanatos and receptor-interacting protein 1/3 (RIP1/3) kinase-induced necroptosis in the pathophysiological course of acute ischemic intestinal injury.. Anesthetized adult male Sprague-Dawley rats were subjected to superior mesenteric artery occlusion consisting of 1.5 h of ischemia and 6 h of reperfusion. The PARP-1-specific inhibitor PJ34 (10 mg/kg) and the RIP1-specific inhibitor Necrostatin-1 (1 mg/kg) were intraperitoneally administered 30 min before the induction of ischemia.. Intestinal I/R was found to result in PARP-1 activation and RIP1/3-mediated necrosome formation. PJ34 or Necrostatin-1 treatment significantly improved the mucosal injury, while the combined inhibition of PARP-1 and RIP1/3 conferred optimal protection of the intestine. Meanwhile, results from terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay showed a decrease in intestinal epithelial cell death. Interestingly, we further showed that PARP-1 might act as a downstream signaling molecule of RIP1 in the process of I/R-induced intestinal injury and that the RIP1/PARP-1-dependent cell death signaling pathway functioned independently of caspase 3 inhibition.. The results of our study provide a molecular basis for combination therapy that targets both pathways of regulated necrosis (parthanatos and necroptosis), to treat acute intestinal I/R-induced intestinal epithelial barrier disruption.

Topics: Animals; Apoptosis; Disease Models, Animal; Drug Therapy, Combination; Epithelial Cells; Humans; Imidazoles; Indoles; Intestinal Mucosa; Male; Necrosis; Phenanthrenes; Poly (ADP-Ribose) Polymerase-1; Protein Serine-Threonine Kinases; Rats; Rats, Sprague-Dawley; Receptor-Interacting Protein Serine-Threonine Kinases; Reperfusion Injury; Signal Transduction; Treatment Outcome

Stroke is the leading cause of neurological disability in humans. Middle cerebral artery occlusion (MCAO) followed by reperfusion is widely accepted to mimic stroke in basic medical research. Triptolide is one of the major active components of the traditional Chinese herb Tripterygium wilfordii Hook F, and has been reported to have potent anti-inflammatory and immunosuppressive properties. Since its preclinical effects on stroke were still unclear, we decided to study the effects of Triptolide on focal cerebral ischemia/reperfusion injury in this study. The results showed that Triptolide treatment significantly attenuates brain infarction volume, water content, neurological deficits, and neuronal cell death rate, which were increased in the MCAO model rats. Immunohistochemistry was used to analyze the expression of glial fibrillary acidic protein (GFAP), Cyclooxygenase-2 (COX-2), inducible nitric oxide (iNOS), and NF-κB in the ischemic brains. The administration of Triptolide showed down-regulation of the iNOS, COX-2, GFAP, and NF-κB expression in MCAO rats. It also increased the expression of bcl-2, and suppressed levels of bax and caspase-3 compared with the MCAO group. Our findings revealed that Triptolide exerts its neuroprotective effects against inflammation with the involvement of inhibition of NF-κB activation.

Topics: Animals; Anti-Inflammatory Agents; Antineoplastic Agents, Alkylating; Apoptosis; Blotting, Western; Cyclooxygenase 2; Disease Models, Animal; Diterpenes; Epoxy Compounds; Glial Fibrillary Acidic Protein; Immunoenzyme Techniques; In Situ Nick-End Labeling; Infarction, Middle Cerebral Artery; Male; Neuroprotective Agents; NF-kappa B; Nitric Oxide; Phenanthrenes; Rats; Rats, Sprague-Dawley; Real-Time Polymerase Chain Reaction; Reperfusion Injury; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Severity of Illness Index; Signal Transduction

Inactivation of poly(ADP-ribose) polymerase 1 (PARP1) has been found to be protective in several disease models; however, the role of PARP1 in acute kidney injury-induced interstitial fibrosis has not been studied. Herein, we tested whether PARP1 inactivation by treatment with PJ34 (a PARP1 inactivator; 10 mg/kg body weight/day, intraperitoneal implantation of a miniosmotic pump at 2 days after the onset) contributed to the decrease in interstitial fibrosis induced by ischemia-reperfusion injury (IRI) in mouse kidneys. IRI increased PARP1 activation represented by poly(ADP-ribose) expression from 4 to 16 days postinjury, whereas treatment with PJ34 at 2 days after the onset efficaciously abolished the increase in PARP1 activation at 4, 8 and 16 days after IRI. Pharmacological inactivation of PARP1 significantly reduced interstitial fibrosis as represented by the collagen deposition and transforming growth factor-β1 level at 8 and 16 days after IRI. Consistent with collagen deposition, myofibroblast activation represented by α-smooth muscle actin expression was also reduced by PARP1 inactivation at 8 and 16 days after IRI. Furthermore, IRI enhanced macrophage influx, but PARP1 inactivaton remarkably reduced macrophage influx for 4 through 16 days after the injury. Among the chemoattractants for monocytes/macrophages and neutrophils, monocyte chemotactic protein-1 (MCP-1) production in IRI kidneys was significantly reduced by PARP1 inactivation from 4 to 16 days postinjury. These data demonstrate that PARP1 activation contributes to IRI-induced MCP-1 production and in turn to macrophage influx, resulting in the promotion of interstitial fibrosis.

Topics: Actins; Acute Kidney Injury; Animals; Chemokine CCL2; Collagen; Enzyme Activation; Enzyme Inhibitors; Fibrosis; Macrophages; Male; Mice; Mice, Inbred C57BL; Myofibroblasts; Phenanthrenes; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Reperfusion Injury

Triptolide (TP), a naturally derived compound, is proven effective in the treatment of nephritis and chronic allograft nephropathy. However, the severe multiorgan toxicity greatly limited it from further clinic use. 2-Glucosamine was demonstrated as a potential targeting ligand that could specifically interact with megalin receptors highly expressed in renal proximal tubules. In this study, 2-glucosamine was employed as a glycosyl donor while triptolide the acceptor to afford a nonhydrolyzable triptolide derivative-triptolide aminoglycoside (TPAG). The kidney-targeting efficiency, pharmacodynamic properties and safety of TPAG were thus evaluated. TPAG displayed 6.94-fold of AUC(0-t, kidney) and 13.96-fold of MRT(0-t, kidney) compared to TP. Additionally, TPAG presented improved protective effect against renal ischemia/reperfusion injury. Compared to TP's multiorgan toxicity, TPAG showed minimum toxicity toward the kidney and genital systems, and greatly lowered toxicity in the liver and immune systems. In sum, our study presented an alternative structure modification of triptolide with improved safety and efficacy profiles.

Topics: Aminoglycosides; Animals; Diterpenes; Epoxy Compounds; Glucosamine; Kidney; Kidney Tubules, Proximal; Liver; Male; Mice; Molecular Structure; Phenanthrenes; Rats, Sprague-Dawley; Reperfusion Injury; Testis; Toxicity Tests

The effects of sodium tanshinone IIA sulfonate (STS) on coronary no-reflow (CNR) relevant to microvascular obstruction (MVO) remain unknown. Studies had shown that fibrinogen-like protein 2 (FGL2) expressed in microvascular endothelial cells (MECs) is a key mediator in MVO. Thus, we aimed to elucidate the roles of STS in CNR and relations between STS and FGL2.. Myocardial ischemia/reperfusion was selected to represent CNR model. The no-reflow zone and infarct area were assessed using Thioflavin S and TTC staining, and cardiac functional parameters were detected using echocardiography. Western blot was used to detected FGL2 level, fibrin level, protease-activated receptor-1 (PAR-1) activation and inflammation cells infiltration. FGL2 and inflammation cells were also identified by IHC. Microthrombus was detected by Carstairs' and MSB staining. We also detected the roles of STS on FGL2 expression, thrombin generation, phospho-Akt and NF-κB levels in MECs.. Upon treatment with STS in CNR model, the no-reflow and infarct areas decreased significantly and cardiac function improved. The FGL2 expression was inhibited by STS in vivo as well as in vitro with thrombin generation inhibition. In addition, STS up-regulates Akt phosphorylation and suppressed NF-κB expression in activated MECs. Furthermore, fibrin deposition, PAR-1 activation and inflammatory response were inhibited with STS administration in CNR model.. Our results displayed a novel pharmacological action of STS on CNR. STS is able to ameliorate CNR through inhibition of FGL2 expression mediated by Akt and NF-κB pathways as well as prevention of MVO by suppressing fibrin deposition and inflammation.

Topics: Animals; Coronary Circulation; Disease Models, Animal; Down-Regulation; Endothelial Cells; Fibrin; Fibrinogen; Male; No-Reflow Phenomenon; Phenanthrenes; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Receptor, PAR-1; Reperfusion Injury; Signal Transduction

Ischemia-reperfusion (I/R) injury after lung transplantation causes alveolar damage, lung edema, and acute rejection. Poly(adenosine diphosphate-ribose) polymerase (PARP) is a single-stranded DNA repair enzyme that induces apoptosis and necrosis after DNA damage caused by reactive oxygen species. We evaluated tissue protective effects of the PARP inhibitor (PARP-i) PJ34 against pulmonary I/R injury.. Rats (total n=45) underwent a thoracotomy with left hilar isolation and saline administration (sham group) or thoracotomy with hilar clamping and saline administration (I/R group) or PJ34 administration (PARP-i group). Parameters were measured for 7 days after reperfusion.. Pathologic analysis revealed that reperfusion injury was drastically suppressed in the PARP-i group 2 days after reperfusion. Terminal deoxynucleotide transferase-mediated deoxyuridine triphosphate nick-end labeling-positive cells were significantly decreased in the PARP-i group compared to the I/R group (P<0.05). Accordingly, the wet-to-dry lung ratio in the I/R group was significantly higher compared with the PARP-i group (P=0.025). Four hours after reperfusion, serum tissue necrosis factor-α and interleukin-6 were significantly suppressed in the PARP-i group compared with the I/R group (P<0.05). Serum derivatives of reactive oxygen metabolites increased quickly and remained high in the I/R and PARP-i groups from 4 hr until 7 days after reperfusion. Interestingly, the serum biologic antioxidant potential in the PARP-i group was significantly higher than that in the I/R group from day 2 until day 7.. The PARP-i decreased inflammation and tissue damage caused by pulmonary I/R injury. These beneficial effects of the PARP-i may be correlated with its antioxidative efficacy.

Topics: Adenosine Triphosphate; Animals; Antioxidants; Apoptosis; DNA Repair; Inflammation; Interleukin-6; Lung; Lung Transplantation; Male; Oxidative Stress; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Rats; Rats, Wistar; Reactive Oxygen Species; Reperfusion Injury; Thoracotomy; Time Factors; Tumor Necrosis Factor-alpha

Cryptotanshinone (CT), isolated from the dried roots of Salvia militorrhiza, has been reported to have protective effects on myocardial and cerebral ischemia/reperfusion (I/R) injury both in vitro and in vivo. However, its effects and underlying mechanism on hepatic I/R injury remain unclear. To investigate its effects on hepatic I/R injury, thirty male Sprague-Dawley rats were randomized into 3 groups: a sham group, a vehicle-treated hepatic I/R group and a CT-treated (50 mg/kg) group. The hepatic I/R and CT-treated groups were subjected to 60 min of normothermic ischemia of the left lateral lobe of the liver, followed by 4 h of reperfusion. The animals were then sacrificed to collect the serum and the left liver lobe for assay. Hepatic function was protected, as evidenced by significantly reduced alanine aminotransferase (ALT), aspartate aminotransferase (AST) and malondialdehyde (MDA) levels in the CT-treated group as compared with I/R group. The terminal deoxynucleotidyl transferase deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) demonstrated significantly decreased apoptosis in the CT-administration animals. Western blotting demonstrated upregulation of the proapoptotic protein Bcl-2, as well as decreased levels of the activated form of caspase-3 and the cleaved form of its substrate, poly(ADP-ribose) polymerase (PARP) in the CT-treated group compared with those of the I/R group. In addition, the phosphorylation of c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinases (MAPKs) was inhibited by CT. Our data suggest that CT attenuates hepatic I/R injury by inhibiting the intrinsic pathway of apoptosis, mediated partly through the inhibition of JNK and p38 MAPK phosporylation.

Topics: Alanine Transaminase; Animals; Apoptosis; Aspartate Aminotransferases; bcl-2-Associated X Protein; JNK Mitogen-Activated Protein Kinases; Liver; Male; Malondialdehyde; Mitochondria; p38 Mitogen-Activated Protein Kinases; Phenanthrenes; Protective Agents; Proto-Oncogene Proteins c-bcl-2; Rats, Sprague-Dawley; Reperfusion Injury; Superoxide Dismutase

To observe effects of Tanshinone- II A sulfonate on expression of Nuclear factor-kappaB (NF-kappaB), Vascular Cell Adhesion Molecule-1 (VCAM-1) and hemorrheology during spinal cord ischemia reperfusion injury,and explore the function and mechnism.. Fifty-four New Zealand rabbits (aged 3 months,weighted 2.0 +/- 0.2 kg) were randomly divided into 6 in sham group (lumbar artery were separated in operation,0.8 ml/kg saline were injected at 0.5 h before and after operation), 24 in ischemia group ( lumbar artery were clipped after seperation, and the same dose of saline), 24 in Tanshinone group (lumbar artery were clipped after seperation, and the same dose of Tanshinone- II A sulfonate) . Abdomincal aorta blood were drawed after treatment respectively at 0.5 h, 1 h, 4 h and 8 h, and tesetd whole blood viscosity [high cut (mpa.s)/150(l/s), middle cut (mpa.s)/60(l/s) and low cut (mpa.s)/10(l/s)], capillary plasma viscosity, red cell aggregation index, rigid index, deformation index and electrophoresis index. Spinal cord tissues were divided into two sections,one fixed in 4% paraformaldehyde, another stored in liquid nitrogen. Immunohistochemical method and ELISA were used to test change of content of NF-kappaB and VCAM-1.. 1) The expression of NF-kappaB in Tanshinone group were lowest, and in ischemia group were highest. 2) Compared with sham group, VCAM-1 in ischemia group at different time were obviously increased,especially at 0.5, 1 and 4 h (P<0.01), and had meaning at 8 h (P<0.05). Compare between Tanshinone group and ischemia group, VCAM-1 at 0.5 h were obviously decreased (P<0.01), and had meaning at 1 h, 4 h and 8 h (P<0.05). 3) There were no postive vasvular expression in sham group, and at 0.5 h in Tanshinone group and ischemia group. The highest postive vasvular expression in ischemia group were at 1 h, 4 h and 8 h, and had significant meaning at 1 h and 4 h between ischemia group and Tanshinone group (P<0.05), and 8 h were obviously most. 4) The whole blood viscosity in ischemia group at 10 s(-1), 60 s(-1), 150 s(-1) were highest, and capillary viscosity increased (P<0.05 or P<0.01). While capillary viscosity, red cell aggregation index, figid index, deformation index in Tanshinone group decreased obviously (P<0.01).. Tanshinone-II A sulfonate can relieve spinal cord ischemia reperfusion injury by regulating expression of NF-kappaB, VCAM-1, decreasing whole blood viscosity, capillary plasma viscosity, red cell aggregation index, rigid index, and improve hemorhelogy.

Topics: Animals; Female; Gene Expression Regulation; Hemorheology; Male; NF-kappa B; Phenanthrenes; Rabbits; Reperfusion Injury; Spinal Cord; Vascular Cell Adhesion Molecule-1

Hepatic I/R injury is unavoidable in liver transplantation and surgery. This remains a significant problem in surgical procedures. The purpose of this study was to investigate the effects of triptolide on liver ischemia/reperfusion (I/R) injury and related mechanisms in mice.. Male C57BL/6 mice were randomized into four groups: (1) sham group; (2) sham-triptolide group; (3) I/R group; and (4) I-R/triptolide group. Ninety minutes of warm ischemia was induced and flow by 24 h reperfusion. Serum alanine aminotransferase and aspartate aminotransferase were assayed, pathologic alterations and (NF)-κB p65 immunohistochemistry were observed. Liver malondialdehyde (MDA) level, activity of endogenous antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activities, and activity of neutrophil accumulation marker myeloperoxidase (MPO) were measured. TNF-α, IL-6, and IL-1β mRNA were detected by RT-PCR, whereas nuclear factor (NF)-κB p65 and IκBα were assessed with Western blotting.. Plasma aminotransferase activity was higher in the I/R group than in the I/R-triptolide group. MDA level and neutrophil infiltration were also markedly reduced, while SOD, CAT, and GSH-Px levels increased in I/R-triptolide group compared with I/R group. In group 4, histopathologic changes were significantly attenuated in triptolide-treated livers. In comparison with group 3, triptolide reduced NF-κB p65 nuclear and IκBα expression, and effectively suppressed pro-inflammatory cytokine level during the I/R.. These results suggest that triptolide has protective effects against hepatic I/R injury. Its mechanisms might be related to reduction of oxidative stress and neutrophil infiltration and inhibition NF-κB p65 activity.

Topics: Alanine Transaminase; Animals; Aspartate Aminotransferases; Diterpenes; Epoxy Compounds; I-kappa B Proteins; Immunosuppressive Agents; Interleukin-1beta; Interleukin-6; Liver Diseases; Male; Malondialdehyde; Mice; Mice, Inbred C57BL; Neutrophils; NF-kappa B; NF-KappaB Inhibitor alpha; Oxidative Stress; Peroxidase; Phenanthrenes; Reperfusion Injury; RNA, Messenger; Tumor Necrosis Factor-alpha

Ischemia and reperfusion have been identified as a complex cascade of inflammatory mediators that are involved in the pathogenesis of hepatic injury. Triptolide (diterpenoid triepoxide), was extracted from a purified component of a traditional Chinese Medicine, Tripterygium wilfondii Hook F. Currently, triptolide has been shown to have anti-inflammatory, immunosuppressive, and antineoplastic activity. Accumulated data have shown that Th17 cells might contribute to the pathogenesis of liver diseases. Triptolide has been shown to reduce interleukin (IL)-17 expression in inflammatory bowel disease and arthritis. However, the role of triptolide in liver ischemia/reperfusion (I/R) and whether it can attenuate injury and the potential mechanism have not been investigated. Mice were treated with triptolide (0.1mg/kg) for 1 week or IL-17 antibody (50 μg/mouse) 2 days before ischemic insult. Partial warm ischemia was produced in the hepatic lobes of C57BL/6 mice for 90 min, followed by various periods of reperfusion. We demonstrated that IL-17 was involved in the inflammatory response to hepatic I/R injury, and that triptolide inhibited IL-17 generation and suppressed neutrophil migration after liver I/R injury through downregulation of signal transducer and activator of transcription 3 (STAT3) transcription. Also, triptolide pretreatment protected the liver from warm I/R injury, at least in part, mediated by the upregulation of Foxp3 expression. These results could pave the way for the use of triptolide as a novel agent to attenuate I/R injury.

Topics: Animals; Antibodies, Monoclonal; Cell Movement; Disease Models, Animal; Diterpenes; Epoxy Compounds; Forkhead Transcription Factors; Gene Expression Regulation; Humans; Interleukin-17; Liver; Liver Diseases; Mice; Mice, Inbred C57BL; Neutrophils; Phenanthrenes; Phytotherapy; Reperfusion Injury; STAT3 Transcription Factor; Tripterygium

Several experiments were designed to determine whether the systemic, postischemic administration of PJ34,which is a poly-adenosine diphosphate (ADP)-ribose polymerase inhibitor, decreased tissue injury and inflammation after hind-limb ischemia reperfusion (I/R).. C57BL6 mouse limbs were subjected to 1.5 h ischemia followed by 24-h reperfusion. The treatment group (PJ) received intraperitoneal PJ34 (30 mg/kg) immediately before reperfusion, as well as 15 min and 2 h into reperfusion. The control group (CG) received lactated Ringer's alone at the same time intervals as PJ34 administration. The skeletal muscle levels of adenosine triphosphate (ATP), macrophage inflammatory protein-2 (MIP-2), keratinocyte derived chemokine (KC), and myeloperoxidase (MPO) were measured. Quantitative measurement of skeletal muscle tissue injury was assessed by microscopic analysis of fiber injury.. ATP levels were higher in limbs of PJ versus CG mice (absolute ATP: 4.7 +/- 0.35 vs 2.3 +/- 0.15-ng/mg tissue, P = .002). The levels of MIP-2, KC, and MPO were lower in PJ versus CG mice (MIP-2: 1.4 +/- 0.34 vs 3.67 +/- 0.67-pg/mg protein, P = .014; KC: 4.97 +/- 0.97 vs 12.65 +/- 3.05-pg/mg protein, P = .037; MPO: 46.27 +/- 10.53 vs 107.34 +/- 13.58-ng/mg protein, P = .008). Muscle fiber injury was markedly reduced in PJ versus CG mice (4.25 +/- 1.9% vs 22.68 +/- 3.0% total fibers, P = .0004).. Systemic postischemic administration of PJ34 preserved skeletal muscle energy levels, decreased inflammatory markers, and preserved tissue viability post-I/R. These results support PARP inhibition as a viable treatment for skeletal muscle I/R in a clinically relevant post hoc scenario.

Topics: Adenosine Triphosphate; Animals; Chemokine CXCL1; Chemokine CXCL2; Disease Models, Animal; Hindlimb; Ischemia; Male; Mice; Mice, Inbred C57BL; Muscle, Skeletal; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Reperfusion Injury

Disruption of blood-brain barrier (BBB) and edema formation play a key role in the development of neurological dysfunction after cerebral ischemia. In this study, the effects of Tanshinone IIA (Tan IIA), one of the active ingredients of Salvia miltiorrhiza root, on the BBB and brain edema after transient middle cerebral artery occlusion in rats were examined. Our study demonstrated that Tan IIA reduced brain infarct area, water content in the ischemic hemisphere. Furthermore, Tan IIA significantly decreased BBB permeability to Evans blue, suppressed the expression of intercellular adhesion molecule-1 (ICAM-1), matrix metalloproteinase-9 (MMP-9), inhibited the degradation of tight junction proteins zonula occludens-1 (ZO-1) and Occludin. These results demonstrated that Tan IIA was effective for attenuating the extent of brain edema formation in response to ischemia injury in rats, partly by Tan IIA's protective effect on the BBB. Our results may have implications in the treatment of brain edema in cerebral ischemia.

Topics: Abietanes; Animals; Blood-Brain Barrier; Brain; Edema; Evans Blue; Infarction, Middle Cerebral Artery; Intercellular Adhesion Molecule-1; Ischemic Attack, Transient; Matrix Metalloproteinase 9; Membrane Proteins; Occludin; Permeability; Phenanthrenes; Phytotherapy; Plant Extracts; Plant Roots; Rats; Reperfusion Injury; Salvia miltiorrhiza; Tight Junctions; Water

To investigate the in vitro effect of tanshinone IIA on leukocyte-associated hypoxia-reoxygenation injury of human brain-blood barrier (BBB), we established the BBB model by culturing purified primary human brain microvascular endothelial cells (HBMVEC) to confluence on cell culture insert. BBB was identified by tight junction, transendothelial electrical resistance (TEER) and the permeability of BBB to horseradish peroxidase (HRP). The effect of tanshinone IIA on the permeability of BBB was tested at 2 h after hypoxia and 1h after reoxygenation with or without the supernatants of activated leukocytes. The effect of tanshinone IIA on leukocytes activation was analyzed by detection of MMP-9, cytokines and reactive oxygen species. The results showed that BBB formed by confluent HBMVECs had no cellular gap. Immunofluorescent staining for ZO-1 confirmed that the cells were connected by tight junction. Moreover, the BBB model had a higher TEER and a lower permeability for HRP than confluent HUVECs. The permeability of BBB for HRP was enhanced by hypoxia-reoxygenation and further greatly enhanced by adding the supernatants of activated leukocytes before reoxygenation. But such an effect was reversed by addition of tanshinone IIA before hypoxia. Moreover, tanshinone IIA could decrease the levels of MMP-9, TNF-α, IL-1α, IL-2, IFN-γ and reactive oxygen species in leukocytes. In conclusion, tanshinone IIA can protect BBB against leukocyte-associated hypoxia-reoxygenation injury by attenuating the activation of leukocytes and inhibiting the injury effects of leukocytic products. Tanshinone IIA may be a novel therapeutic agent for cerebral ischemia-reperfusion injury.

Topics: Abietanes; Blood-Brain Barrier; Culture Media, Conditioned; Electric Impedance; Endothelial Cells; Humans; Hypoxia; Leukocytes; Oxygen; Permeability; Phenanthrenes; Reperfusion Injury

We have synthesized a renal-specific drug carrier, 14-succinyl triptolide-lysozyme (TPS-LZM) conjugate for targeted delivery of TP to the PTECs. TPS-LZM could be taken up by HK-2 cells, free TP would be degraded and released, mainly from basolateral side of the cells. Compared with TP, the overall targeting efficiency (TE) of TPS-LZM was significantly enhanced from 11.74% to 95.54% and its MRT was moderately prolonged from 3.08h to 4.10h. At very low concentration, TPS-LZM could significantly reverse the disease progression in renal ischemia-reperfusion (I/R) injury animal models, while the mixture of free TP and LZM was ineffective. Further, TPS-LZM conjugate presented much lower hepatotoxicity (0.78 folds lower than TP) and no adverse effect on the immune (1.13 folds higher than TP) and genital system. Thus, TPS-LZM represents a very effective drug candidate for specific treatment of immunological renal diseases with low adverse side effect.

Topics: Animals; Diterpenes; Drug Carriers; Epithelial Cells; Epoxy Compounds; Humans; Kidney; Kidney Diseases; Kidney Tubules, Proximal; Male; Molecular Structure; Muramidase; Phenanthrenes; Rats; Rats, Wistar; Reperfusion Injury; Succinates; Tissue Distribution

Visceral ischemia-reperfusion injury (VI) contributes to adverse outcomes following the repair of thoracoabdominal aneurysms. Experiments were designed to determine whether a poly-adenosine diphosphate-ribose polymerase (PARP) inhibitor modulates indexes of metabolic function (mitochondrial activity), inflammatory cell activation, and tissue inflammation (lipopolysaccharide receptor CD14 messenger ribonucleic acid) following VI.. 129S1/SvImj mice were subjected to thoracic aortic occlusion followed by 48 hours of reperfusion. Normal saline was administered to 25 untreated control mice and PJ34 to 21 mice before and immediately after thoracic aortic ischemia-reperfusion. Sham mice (n = 13) underwent median sternotomy alone. At 48 hours, all animals were euthanized and tissues harvested for quantitative analysis.. PJ34 improved intestinal (P < .05) but not hepatic mitochondrial activity following reperfusion. CD14 messenger ribonucleic acid levels in liver (P < .004), kidney (P < .003), and spinal cord (P < .03) tissue were less in PJ34-treated mice.. PJ34 preserved the metabolic function of intestinal but not hepatic tissue during reperfusion. PJ34 uniformly decreased the expression of an important marker of inflammatory cell activation and tissue inflammation in visceral tissue following VI. PARP inhibitors may serve as a therapeutic modality to abrogate the stress response to VI.

Topics: Animals; Aorta, Thoracic; Intestinal Mucosa; Kidney; Lipopolysaccharide Receptors; Liver; Male; Mice; Mitochondria; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Polymerase Chain Reaction; Reperfusion Injury; RNA, Messenger; Spinal Cord

The author evaluates an experimental model of hepatic resection. The goal of the study was to investigate ischaemic tolerance during liver resection. In this model, we also investigated the protective effects of ischemic preconditioning (IP), a PARP enzyme inhibitor (PJ-34), and an antioxidant glutamine (Gln). Inbred male Wistar rats were used for the experiments. Complete segmental ischaemia of the liver was achieved with or without using IP, PJ-34 or Gln pretreatment before the ischaemia. Microcirculation was monitored using laser Doppler flowmeter (LDF) throughout the ischaemia-reperfusion period. Required standardizations and mathematical analyses were performed in order to validate statistical analysis. Histological changes, immunohistochemistry (TUNEL, caspase), liver enzymes, bilirubin, and TNF-alpha levels were all measured simultaneously.. 30 minutes ischaemia was well tolerated by the liver and IP does not cause further improvement. 45 to 60 minutes ischaemia resulted in serious microcirculatory changes during reperfusion. 90 minutes ischaemia was unequivocally intolerable. The liver injury and microcirculatory changes caused by 45 or 60 minutes of ischemia could be reduced with IP. Improvement was observed both on histology and in survival rates. After 45 and 60 minutes, IP + I-R caused a significant decline in the TNF-alpha levels. IP, PJ-34 PARP-inhibitor and glutamine pretreatment prior to 60 minutes of ischaemia resulted in an improvement of microcirculation. Immunohistochemistry for activated caspase-3 demonstrated significantly higher apoptosis rates in the PJ-34 PARP-inhibitor and glutamine-pretreated groups, in contrast to the sham and I-R groups. Antioxidant levels in the serum and in the liver homogenates showed significant improvement in the IP and glutamine-pretreated groups.

Topics: Animals; Antioxidants; Apoptosis; Caspase 3; Glutamine; Hepatectomy; Immunohistochemistry; In Situ Nick-End Labeling; Ischemic Preconditioning; Laser-Doppler Flowmetry; Liver; Liver Circulation; Liver Function Tests; Male; Microcirculation; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Rats; Rats, Wistar; Reperfusion Injury; Time Factors; Tumor Necrosis Factor-alpha

The nuclear enzyme poly(ADP-ribose) polymerase (PARP) is activated by oxidative stress and plays a significant role in postischemic brain injury. We assessed the contribution of PARP activation to the blood-brain barrier (BBB) disruption and edema formation after ischemia-reperfusion. In male Wistar rats, global cerebral ischemia was achieved by occluding the carotid arteries and lowering arterial blood pressure for 20 mins. The animals were treated with saline or with the PARP inhibitor N-(6-oxo-5,6-dihydrophenanthridin-2-yl)-N, N-dimethylacetamide.HCl (PJ34); (10 mg/kg, i.v.) before ischemia. After 40 mins, 24, and 48 h of reperfusion, the permeability of the cortical BBB was determined after Evans Blue (EB) and Na-fluorescein (NaF) administration. The water content of the brain was also measured. The permeability of the BBB for EB increased after ischemia-reperfusion compared with the nonischemic animals after 24 and 48 h reperfusion but PARP inhibition attenuated this increase at 48 h (nonischemic: 170+/-9, saline: 760+/-95, PJ34: 472+/-61 ng/mg tissue). The extravasation of NaF showed similar changes and PJ34 post-treatment attenuated the permeability increase even at 24 h. PARP inhibition decreased the brain edema seen at 48 h. Because PARP has proinflammatory properties, the neutrophil infiltration of the cortex was determined, which showed lower values after PJ34 treatment. Furthermore, PJ34 treatment decreased the loss of the tight junction protein occludin at 24 and 48 h. The inhibition of PARP activity accompanied by reduced post-ischemic BBB disturbance and decreased edema formation suggests a significant role of this enzyme in the development of cerebral vascular malfunction

Topics: Animals; Blood-Brain Barrier; Blotting, Western; Brain; Brain Edema; Capillary Permeability; Enzyme Activation; Hypoxia-Ischemia, Brain; Intercellular Adhesion Molecule-1; Male; Membrane Proteins; Occludin; Peroxidase; Phenanthrenes; Poly(ADP-ribose) Polymerases; Rats; Rats, Wistar; Reperfusion Injury; Reverse Transcriptase Polymerase Chain Reaction

Ischemia-reperfusion (I-R) injury during liver resection leads to the production of toxic free radicals and oxidants that influence the microcirculation. DNA single-strand breaks can be induced by these reactive species. In response to excessive DNA damage, PARP [poly(ADP-ribose) polymerase] becomes overactivated, which can lead to cellular ATP depletion and cell death. The aim of our study was to evaluate whether PARP is expressed in post-ischemic liver, and to examine the effect of the administration of PJ-34 PARP inhibitor on liver function, histopathology, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) reaction, and the oxidative state of the liver after injury.. Male Wistar rats (weighing 250 g) underwent 60 min of normothermic, segmental liver ischemia followed by 30 min of reperfusion. The animals (n = 45) were divided into three groups: sham operated; I-R (control) treated with saline; and PJ-34 pre-treated (10 mg/kg i.v.). Hepatic microcirculation was monitored by a laser Doppler flowmeter. The reperfusion was characterized as the integral of the reperfusion area (RA) and the maximal plateau (PM). Histological alterations, TUNEL-reaction, serum, and liver tissue antioxidant levels, as well as serum ALT and AST levels were measured.. Upon reperfusion, the PJ-34 group had significantly (P < 0.05) higher flow rates than control groups (PM(PJ-34): 58%, PM(control): 37%; RA(PJ-34.): 48%, RA(control): 25%). At the end of the 30 min reperfusion, PJ-34 resulted in significantly (P < 0.05) lower serum ALT and AST levels and chemiluminescent intensity (free radicals) of the liver. The liver's free SH-group concentration and H-donor ability of the plasma was elevated in the PARP-inhibitor treated group. Positive staining for TUNEL, after PJ-34 pre-treatment was significantly increased (P < 0.05); in contrast, the control tissues were less positively stained for TUNEL but necrotic tissue was abundant.. PARP plays a pathogenetic role in the deterioration of the hepatic microcirculation and promotes hepatocellular necrosis in liver reperfusion injury.

Topics: Alanine Transaminase; Animals; Antioxidants; Apoptosis; Glutamyl Aminopeptidase; Liver; Male; Microcirculation; Models, Animal; Necrosis; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Rats; Rats, Wistar; Reactive Oxygen Species; Regional Blood Flow; Reperfusion Injury

To investigate the protective effect of triptolide (TRI) on ischemia/reperfusion-induced injury of transplanted rabbit lungs and to investigate the mechanisms underlying the actions of TRI.. We established the rabbit lung transplantation model and studied lung injury induced by ischemia/reperfusion and the inhibitory effect of TRI on NF-kappaB. The severity of lung injury was determined by a gradual decline in PvO2, the degree of lung edema, the increase in the myeloperoxidase (MPO) activity, and the ultrastructural changes of transplanted lungs. The activation of NF-kappaB was measured by immunohistochemistry. The increase in intercellular adhesion molecule-1 (ICAM-1), which is the target gene of NF-kappaB, was evaluated by ELISA.. After reperfusion, there was a gradual decline in the PvO2 level in the control group (group I). The level of PvO2 in the group treated with lipopolysaccharide (group II) was significantly decreased, whereas that of the group treated with TRI (group III) was markedly improved (P<0.01). In group III, the activity of MPO was downregulated, and the pulmonary edema did not become severe and the ultrastructure of the donor lung remained normal. The activity of NF-kappaB and the expression of ICAM-1 was significantly increased in the donor lungs. TRI blocked NF-kappaB activation and ICAM-1 expression.. The effects of TRI on reducing injury to donor lungs induced by ischemia/reperfusion may possibly be mediated by inhibiting the activity of NF-kappaB and the expression of the NF-kappaB target gene ICAM-1. Thus, TRI could be used in lung transplantations for improving the function of donor lungs.

Topics: Animals; Diterpenes; Enzyme-Linked Immunosorbent Assay; Epoxy Compounds; Lung; Lung Transplantation; Models, Animal; NF-kappa B; Phenanthrenes; Rabbits; Reperfusion Injury

Systemic inflammatory responses contribute to mortality after thoracoabdominal aneurysm repair. Poly adenosine diphosphate (ADP) ribose polymerase (PARP) activity is known to modulate inflammation in animal models of injury. The effect of the PARP inhibitor PJ34 and genetic deletion of PARP-1(PARP -/-) on the systemic inflammatory response after thoracic aortic ischemia reperfusion (TAR) is not known.. In one group, all mice were subject to TAR followed by 48 hours of reperfusion. Treated mice (PJ, n=24) were given PJ34 IP; untreated mice (UN, n=41) received normal saline intraperitoneally. The number of mice in each group was selected to have a similar number of survivors by 48 hours. In a second group, sham animals were subjected to mediastinotomy alone (sham, n=10) without TAR, and were compared with mice with deletion of the PARP-1 isoform (PARP-1 -/-, n=11) subjected to TAR. Tissue extracts were assayed for keratinocyte derived chemokine and granulocyte colony stimulating factor. Serum was assayed for interleukin-6.. PJ34 treatment decreased mortality throughout the experimental protocol. There were no mortalities in the sham operated mice or PARP -/- mice subjected to TAR. PJ34 treatment decreased serum levels of interleukin-6 (p=0.01) and hepatic levels of interleukin-6 mRNA when compared with untreated and PARP-/- mice (p < 0.01). Only liver and kidney cytokine levels were decreased by PJ34 treatment (p < 0.05). In PARP-/- mice subjected to TAR, tissue cytokine levels were not different from those in sham mice.. PARP inhibition may represent a novel therapeutic approach to minimizing inflammatory sequelae after TAR.

Topics: Animals; Aorta, Thoracic; Cytokines; Disease Models, Animal; Male; Mice; Phenanthrenes; Poly (ADP-Ribose) Polymerase-1; Poly(ADP-ribose) Polymerase Inhibitors; Poly(ADP-ribose) Polymerases; Reperfusion Injury; Systemic Inflammatory Response Syndrome

To investigate whether tanshinone IIA could improve the effect of UW solution for skeletal muscle preservation and to determine the dose range of tanshinone IIA providing optimal protection during ischemia and reperfusion.. Ischemic rat limbs were perfused with UW solution or UW plus tanshinone IIA (UW+T, 0.05, 0.1, or 0.2 mg/mL) for 0.5 h before reperfusion; controls (I/R) received no perfusion. Serum creatine phosphokinase (CPK), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) were measured pre-ischemia and after reperfusion (2-h, 4-h, and 6-h). Muscle water content, superoxide dismutase (SOD), malondialdehyde (MDA), adenosine triphosphatase (ATPase) were assessed pre-reperfusion and after 6-h reperfusion. Intercellular adhesion molecule-1 (ICAM-1) and apoptosis were detected after 6-h reperfusion. Reperfusion blood flow was monitored during reperfusion period.. UW and UW+T prevented luxury perfusion during reperfusion and inhibited ICAM-1 expression and apoptosis after 6-h reperfusion. Serum CPK, AST, and LDH levels in UW rats were significantly less than those in controls after 2-h reperfusion (no difference, 4-h or 6-h reperfusion). After 4-h ischemia, there were significant differences in water content, MDA, SOD, and ATPase between UW and controls, but no difference after 6-h reperfusion. All tests with UW+T rats were significantly different from control results at corresponding durations. Higher tanshinone doses improved results.. UW plus tanshinone IIA increased protection against I/R injury, suggesting that tanshinone IIA has clinical value.

Topics: Abietanes; Adenosine; Adenosine Triphosphatases; Allopurinol; Animals; Apoptosis; Aspartate Aminotransferases; Creatine Kinase; Dose-Response Relationship, Drug; Glutathione; Hindlimb; Insulin; Intercellular Adhesion Molecule-1; L-Lactate Dehydrogenase; Male; Malondialdehyde; Muscle, Skeletal; Organ Preservation Solutions; Phenanthrenes; Raffinose; Random Allocation; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Superoxide Dismutase

PARP stabilizes DNA and modulates inflammation in murine models of sepsis, stroke, and myocardial infarction. Previous studies have shown that systemic PARP inhibition before hindlimb ischemia preserves tissue viability and modulates cytokine synthesis during reperfusion. The purpose of this study was to determine whether intra-muscular (IM) administration of PJ34, a potent inhibitor of PARP, after the onset of acute hindlimb ischemia (post hoc) modulates the local production of inflammatory mediators during ischemia/reperfusion (I/R).. The control tension tourniquet was used to establish unilateral hindlimb ischemia in mice for 3 h followed by 48 h I/R. The treatment group (PJ) received IM PJ34 (10 mg/kg) in the affected hindlimb 90 min into ischemia whereas the control group (UN) received IM saline (150 uL) at the same time point. Skeletal muscle viability (MTT mitochondrial activity), local neutrophil chemoattractant protein (KC), Interleukin 6 (IL-6), Interleukin 1beta (IL-1beta), and Myeloperoxidase (MPO) levels were measured in protein extracts after the reperfusion period.. Muscle viability (102% +/- 10 PJ, 78% +/- 4 UN, P = 0.04), IL-B (21.1 +/- 1.3 PJ, 15.5 +/- 1.0 UN, P = 0.02), and IL-6 levels (16.3 +/- 1.2 PJ, 10.9 +/- 1.4 UN, P = 0.04) after 48 I/R were significantly higher in PJ. KC and MPO levels were higher in PJ but neither reached statistical significance.. Post hoc PJ34 therapy appears to protect skeletal muscle from I/R injury despite increased levels of local cytokines. These initial findings support the role of local post hoc therapy in the treatment of acute limb threatening ischemia suggesting that further study of this novel therapy is warranted.

Topics: Animals; Enzyme-Linked Immunosorbent Assay; Hindlimb; Interleukin-1beta; Interleukin-6; Laser-Doppler Flowmetry; Mice; Muscle, Skeletal; Peroxidase; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Reperfusion Injury

Spinal cord injury (SCI) remains a source of morbidity after thoracoabdominal aortic reconstruction. These studies were designed to determine whether PJ34, a novel ultrapotent inhibitor of the nuclear enzyme poly(adenosine diphosphate ribose) polymerase (PARP) could modulate neurologic injury after thoracic aortic ischemia reperfusion (TAR) in a murine model of SCI.. Forty-one anesthetized male mice were subject to thoracic aortic occlusion (11 minutes) through a cervical mediastinotomy followed by 48 hours of reperfusion (TAR) under normothermic conditions. PJ34-treated mice (PJ, n = 12) were given 10 mg/kg PJ34 intraperitoneally 1 hour before ischemia and 1 hour after unclamping. The control group (UN, n = 21) received normal saline intraperitoneally 1 hour before ischemia and 1 hour after unclamping. Sham animals (n = 10) were subject to thoracic aortic exposure with no aortic clamping and similar intraperitoneal normal saline injections. PARP-1-/- (KO, n = 8) mice were subjected to the same conditions as the UN mice. Blinded observers rated murine neurologic status after TAR by using an established rodent paralysis scoring system. Murine spinal cords were subjected to cytokine (GRO-1) protein analysis as a marker of inflammation and immunohistochemical analysis (hematoxylin-eosin and PAR staining). Paralysis scores (PS) and GRO-1 levels were compared with analysis of variance, and survival data were compared with chi 2 .. Immediately after TAR, UN and PJ mice had severe neurologic dysfunction (PS = 5.8 +/- 0.1 and 4.6 +/- 0.6, respectively; P > .05), which was significantly worse than the KO mice (PS = 1.0 +/- 0.7, P < .001). After 6, 24, and 48 hours KO mice had no discernable neurologic injury (PS = 0). Six hours after TAR, PJ mice significantly improved (PS = 1.1 +/- 0.73, P < .001) and remained improved at 24 (PS = 0.7 +/- 0.6) and 48 hours (PS = 0.6 +/- 0.6). UN mice did not improve their PS, and Sham mice showed no neurologic abnormality at any time during these experiments. The mortality at 48 hours was 0% for PJ and KO mice, 43% for UN (P = .012), and 0% for Sham. GRO-1 levels were significantly decreased in PJ and KO versus UN mice (UN, 583 +/- 119 vs PJ, 5.8 +/- 0 vs KO, 5.3 +/- 1.4 mg/pg; P < .0001). Immunohistochemistry showed evidence of decreased PAR staining and ventral motor neuron injury in PJ mice.. Genetic deletion of PARP or inhibition of its activity (PJ34) rescued neurologic function in mice subjected to TAR. PARP inhibition might represent a novel therapeutic approach for prevention of SCI after TAR.

Topics: Animals; Aorta, Abdominal; Aorta, Thoracic; Immunohistochemistry; Injections, Intraperitoneal; Male; Mice; Motor Neurons; Phenanthrenes; Poly Adenosine Diphosphate Ribose; Poly(ADP-ribose) Polymerase Inhibitors; Protozoan Proteins; Reperfusion Injury; Spinal Cord; Spinal Cord Diseases

Polyadenosine diphosphate-ribose polymerase (PARP) has been implicated as a mediator of inflammation and tissue necrosis in murine models of human stroke and myocardial infarction. This study was designed to determine whether PARP modulates skeletal muscle injury and cytokine-growth factor levels during ischemia-reperfusion.. Prospective controlled animal study.. Medical school-affiliated university hospital.. Mice were divided into 2 groups-treated (PJ) and untreated; all mice were subjected to unilateral hind limb tourniquet ischemia followed by 4 or 48 hours of reperfusion. In treated mice, PJ34, an ultrapotent-specific PARP inhibitor was given immediately before ischemia and prior to reperfusion. A group of PARP-1 knockout mice (PARP-/-) were also subjected to hind limb ischemia followed by 48 hours of reperfusion.. After ischemia-reperfusion, muscle was harvested for measurement of edema, viability, cytokine, and vascular endothelial growth factor content.. The PJ34-treated mice had increased skeletal muscle viability when compared with the untreated mice after 4 and 48 hours of reperfusion (P<.01). Viability between PARP-/- and PJ34-treated mice were similar at 48 hours of reperfusion (P>.05), and it exceeded that of untreated mice (P<.01). Tissue edema was unaltered by PARP inhibition. Tissue levels of cytokine were only different (P<.05) in PJ34-treated vs untreated mice at 48 hours of reperfusion. Vascular endothelial growth factor levels in PJ34-treated mice were markedly reduced when compared with untreated mice only after 4 hours of reperfusion (P<.01), and in PARP-/- mice (P<.01) at 48 hours of reperfusion.. Polyadenosine diphosphate-ribose polymerase modulates skeletal muscle viability, cytokine and vascular endothelial growth factor synthesis during reperfusion. Polyadenosine diphosphate-ribose polymerase inhibition may represent a novel method to modulate skeletal muscle ischemia-reperfusion injury.

Topics: Analysis of Variance; Animals; Chemokines, CXC; Hindlimb; Immunoenzyme Techniques; Mice; Mice, Inbred Strains; Muscle, Skeletal; Phenanthrenes; Poly Adenosine Diphosphate Ribose; Prospective Studies; Reperfusion Injury; Vascular Endothelial Growth Factor A

These experiments sought to evaluate the effects of PJ34, a poly-ADP-ribose polymerase inhibitor, on molecular indices of renal injury, mitochondrial function, tissue thrombosis, and fibrinolysis after thoracic aortic ischemia/reperfusion (TAR).. Forty-three 129S1/SvImj mice were subjected to 11 minutes of TAR followed by 48 hours of reperfusion. Experimental groups included untreated normal saline (NS) controls (UC), (n=15, 0.5 mL NS i.p.) or PJ34 (PJ) (n=17, PJ34 10 mg/kg ip, 1 hour before and after TAR). Sham (SH) mice (n=11) underwent median sternotomy (heparin, NS i.p.) without TAR. Forty-eight hours after TAR or sham operation, kidney mitochondrial activity (using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium [MTT]), D-dimer, and thrombin-antithrombin III (TAT) complex levels were measured. Levels of messenger RNA for neutrophil gelatinase-associated lipocalin (NGAL), a marker for renal injury, were also measured by reverse transcriptase-polymerase chain reaction.. PJ34 improves renal mitochondrial activity after 48 hours of TAR, compared with untreated control animals (UC, 87.6 +/- 2.2%; PJ, 151.4 +/- 9.5%; P < .001). PJ34 did not alter the increase in renal D-dimer levels by 48 hours reperfusion (UC, 1.37 +/- 0.09 U; PJ, 1.1 +/- 0.14 U; SH, 0.82 +/- 0.06 U; P < .05). TAR did not alter renal levels of TAT expression among groups (UC, 0.103 +/- 0.034; PJ, 0.067 +/- 0.008; SH, 0.106 +/- 0.027; P=.619). The incidence of significantly increased NGAL among UC mice was 1415 +/- 823.6 (n=12), compared with 29.6 +/- 20.8 (n=10) in the PJ34-treated group (P < .014).. PJ34 preserves renal mitochondrial activity and decreases steady-state levels of NGAL after TAR. TAR did increase markers of fibrinolysis in renal tissue but their increase did not correlate with renal injury or PJ34 treatment. These studies indicate that PJ34 confers protection against TAR and suggest that PARP may represent a novel target for reducing perioperative renal injury.

Topics: Acute-Phase Proteins; Animals; Antithrombin III; Fibrin Fibrinogen Degradation Products; Kidney; Kidney Diseases; Lipocalin-2; Lipocalins; Male; Mice; Mice, Inbred Strains; Mitochondria; Oncogene Proteins; Peptide Hydrolases; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Reperfusion Injury; RNA, Messenger; Thoracic Arteries

To study effect of triptolide (TL) on neuronal apoptosis in cerebral tissue of rat after ischemia-reperfusion.. Triptolide at dose 0.2 or 0.4 mg x kg(-1) was intraperitoneally injected once a day for 4 d. The focal cerebral ischemia-reperfusion model was established with thread embolism in middle artery before triptolide injection on the fourth day. Neurological deficit score of rats was evaluated; and immunohistochemical techniques were used to count positive cells of express of MPO and TUNEL in cerebraltissue.. Compared with the control group, the deficit of neural function was significantly improved, and the number of infiltrate of neutrophil and neuronal apoptosis in cerebral tissue was remarkably reduced in two TL-treated groups.. The results suggested that TL can inhibit infiltration of neutrophil and decrease the degree of neuronal apoptosis in cerebral tissue.

Topics: Animals; Apoptosis; Brain Ischemia; Diterpenes; Epoxy Compounds; Female; Infarction, Middle Cerebral Artery; Male; Neuroprotective Agents; Neutrophil Infiltration; Phenanthrenes; Plants, Medicinal; Rats; Rats, Wistar; Reperfusion Injury; Tripterygium

Topics: Animals; Enzyme Inhibitors; Phenanthrenes; Poly(ADP-ribose) Polymerase Inhibitors; Reperfusion Injury; Shock