peoniflorin and Reperfusion-Injury

The pathophysiological mechanism of acute kidney injury (AKI) is complicated, and effective drugs are still lacking. Ferroptosis is a newly discovered regulatory cell death mode characterized by the lethal accumulation of iron and reactive oxygen species-(ROS-)-dependent lipid hydroperoxides. In recent years, ferroptosis has been confirmed to be involved in the progression of AKI. Paeoniflorin (PF) is a traditional Chinese medicine that has protective effects on a variety of kidney diseases including AKI. However, the mechanism by which PF attenuates AKI is unclear. We detected that PF attenuated serum biochemical markers, histological damage, ferroptosis and inflammation in a dose-dependent manner in a mouse AKI model with bilateral renal artery ischemia-reperfusion (IR). Hypoxia-reoxygenation (HR)-induced ferroptosis and inflammation was also inhibited by PF in human renal tubular epithelial cells (HK2). RNA sequence analysis revealed that PF inhibited ferroptosis in HK2 cells by upregulating Slc7a11 in the glutathione pathway after HR treatment. PF failed to further protect cells with specific knockdown of Slc7a11 from ferroptosis under HR conditions. Consequently, these data indicated that PF prevention of ferroptosis in AKI requires dependence on Slc7a11. This study provided a scientific basis for the clinical search for drugs to prevent IR induced AKI.

Topics: Acute Kidney Injury; Amino Acid Transport System y+; Animals; Disease Models, Animal; Ferroptosis; Humans; Hypoxia; Inflammation; Ischemia; Mice; Reperfusion Injury

Paeoniae Radix Rubra (PRR), the root of Paeonia lactiflora Pall. or Paeonia veitchii Lynch, has been widely used to promote blood circulation and eliminate blood stasis in Chinese clinical practice, but its effect on cerebral ischemia is still rarely reported.. The present study aimed to assess the potential therapeutic possibilities of the extract of PRR (PRRE) on cerebral ischemia, further exploring the underlying mechanism, and preliminary screening of the corresponding active components.. The neuroprotective effects of PRRE in Sprague-Dawley (SD) rats with middle cerebral artery occlusion (MCAO) injury and mouse hippocampal neuronal cells (HT22 cell line) following oxidative stress were confirmed. The mechanism was investigated using immunohistochemical staining, western blotting, transmission electron microscopy (TEM), and immunofluorescence. The active components of PRRE were analysed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and molecular docking.. PRRE exerts neuroprotective effects against cerebral ischaemic injury by inhibiting ferroptosis and activating autophagy through the PI3K/Akt signalling pathway. This study provides an experimental basis for the potential application of PRRE as a novel therapeutic drug, and PI3K/Akt-associated ferroptosis and autophagy as therapeutic targets for cerebral ischemia.

Topics: Animals; Autophagy; Beclin-1; Brain Ischemia; Chromatography, Liquid; Ferroptosis; Hydrogen Peroxide; Infarction, Middle Cerebral Artery; Mice; Molecular Docking Simulation; Neuroprotective Agents; Phosphatidylinositol 3-Kinase; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Tandem Mass Spectrometry

In this study, the main hypothesis is that paeoniflorin may inhibit some cellular processes such as oxidative stress and inflammation. For this reason, we aimed to investigate the potential protective effects of a natural compound, paeoniflorin, on rat model of ovarian ischemia-reperfusion injury by detecting the oxidative stress parameters and inflammatory process parameters. 42 female Wistar-albino rats were divided into 6 random groups. The rats were subjected to 3-hour ischemia and 3-hour reperfusion process. Then, paeoniflorin at doses of 25, 50 and 100 mg/kg were applied 30 min before the reperfusion. The levels of pro-inflammatory (IL-1-β, IL-6, TNF-α) and anti-inflammatory (IL-10, TGF-β) cytokines were measured by ELISA. Similarly, IL-6, IL-10, TNF-α, NF-κB p65) positivity rates were detected by immunohistochemical staining. Additionally, oxidative stress parameters (MDA, GSH, SOD) were measured by tissue biochemistry. Ischemia-reperfusion injury caused significant increase in the levels of SOD, MDA, TNF-α, IL-1-β, IL-6 and NF-κB p65, while paeoniflorin treatments improved the related parameters in a dose-dependent manner. As a conclusion, our findings support the evidence that paeoniflorin has a potential protective effects on ovarian ischemia-reperfusion injury. Further detailed studies should be performed to shed light the molecular mechanism of these protective effects.

Topics: Animals; Biological Products; Female; Interleukin-1; Interleukin-10; Interleukin-6; NF-kappa B; Ovary; Rats; Rats, Wistar; Reperfusion Injury; Superoxide Dismutase; Tumor Necrosis Factor-alpha

Topics: Acute Kidney Injury; Apoptosis; Humans; Hypoxia; Kelch-Like ECH-Associated Protein 1; NF-E2-Related Factor 2; Oxidative Stress; Reactive Oxygen Species; Reperfusion Injury; Signal Transduction; Superoxide Dismutase

Paeoniflorin (PF) and calycosin-7-glucoside (CG,. To investigate the synergistic effects of PF + CG on ischaemia/reperfusion injury. Male Sprague-Dawley rats were subjected to the middle cerebral artery occlusion/reperfusion (MCAO/R). After MCAO/R for 24 h, rats were randomly subdivided into 5 groups: sham, model (MCAO/R), study treatment (PF + CG, 40 + 20 mg/kg), LY294002 (20 mg/kg), and study treatment + LY294002. Males were given via intragastric administration; the duration of the. PF + CG significantly reduced neurobehavioral outcomes (21%), cerebral infarct volume (44%), brain edoema (1.6%) compared with the MCAO/R group. Moreover, PF + CG increased p-PI3K/PI3K (4.69%, 7.4%), p-AKT/AKT (6.25%, 60.6%) and Bcl-2/BAX (33%, 49%) expression. PF + CG showed a synergistic protective effect against ischaemic brain injury, potentially being a future treatment for ischaemic stroke.

Topics: Animals; Brain Ischemia; Glucosides; Glycogen Synthase Kinase 3 beta; Infarction, Middle Cerebral Artery; Ischemic Stroke; Isoflavones; Male; Monoterpenes; Neuroprotective Agents; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Reperfusion Injury; Stroke

The purpose of this paper was to study the protective effect of paeoniflorin on acute cerebral ischemia. The animal model of cerebral infarction induced by Middle Cerebral Artery Occlusion (MCAO) was blocked by the suture method. Sixty SD rats were randomly divided into the shame group, MCAO group, paeoniflorin (60, 120, 240 mg/kg, respectively) and Nimodipine (NMDP) group (n = 10 per group).. The rats were intragastrically administered immediately after the operation. After 7 days of gavage, the brains were decapitated at 24 h. Hematoxylin and Eosin (HE) staining was used to observe the degree of cell damage in the cerebral cortex of rats. Immunohistochemistry was used to detect silver plating and to observe changes in nerve cells. Rats in the model group showed obvious symptoms of neurological deficits, such as the ischemic morphological changed, the Malondialdehyde (MDA), Lactate Dehydrogenase (LD) content and lactate dehydrogenase (LDH) activity were significantly increased in the ischemic brain tissue, while the Superoxide Dismutase (SOD) activity was decreased.. The decrease in Na+-K+-ATPase activity was significantly lower than that in the sham group. The neurological symptoms and signs of MCAO in the different doses of paeoniflorin group were improved, and the neuronal edema in the cortical area was alleviated. The activities of SOD, LDH and Na+-K+-ATPase were significantly increased, and the contents of MDA and LD were decreased.. Therefore, paeoniflorin could alleviate the degree of tissue damage in rats with acute cerebral infarction, inhabit the formation of free radicals in the brain tissue after ischemia, and reduce the degree of lipid peroxidation. Thus, the degree of cell damage was reduced greatly and a protective effect was showed on cerebral ischemia.

Topics: Acute Disease; Animals; Brain; Cerebral Infarction; Disease Models, Animal; Glucosides; Lipid Peroxidation; Male; Malondialdehyde; Monoterpenes; Neurons; Neuroprotective Agents; Nimodipine; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Sodium-Potassium-Exchanging ATPase; Superoxide Dismutase

Intestinal ischemia-reperfusion (I/R) injury is a common pathological process with high clinical morbidity and mortality. Paeoniflorin, a monoterpene glucoside, is found to have diverse health beneficial effects including autophagy modulation, anti-inflammatory, anti-apoptotic, and anti-oxidative effects. Based on our pre-experiments, we proposed that paeoniflorin could ameliorate intestinal I/R injury and restore autophagy through activating LKB1/AMPK signal pathway. Our proposal was verified using rat intestinal I/R model in vivo and intestinal epithelial cell line (IEC-6 cells) hypoxia/reoxygenation (H/R) model in vitro. Our results showed that paeoniflorin pretreatment exerted protective effects in rat intestinal I/R injury by reducing intestinal morphological damage, inflammation, oxidative stress, and apoptosis. Paeoniflorin restored H/R-impaired autophagy flux by up-regulating autophagy-related protein p62/SQSTM1 degradation, LC3II and beclin-1 expression, and autophagosomes synthesis without significantly affecting control IEC-6 cells. Paeoniflorin pretreatment significantly activated LKB1/AMPK signaling pathway by reversing the decreased LKB1 and AMPK phosphorylation without affecting total LKB1 both in vivo and in vitro. LKB1 knockdown reduced AMPK phosphorylation, suppressed LC3II and Beclin-1 level, and decreased the degradation of SQSTM/p62, and the knockdown weakened the effects of paeoniflorin in restoring the impaired autophagy flux in H/R injured IEC-6 cells, suggesting that paeoniflorin mitigated the intestinal I/R-impaired autophagy flux by activating LKB1/AMPK signaling pathway. Our study may provide valuable information for further studies.

Topics: AMP-Activated Protein Kinase Kinases; AMP-Activated Protein Kinases; Animals; Apoptosis; Autophagy; Beclin-1; Cell Line; Glucosides; Intestines; Male; Monoterpenes; Oxidative Stress; Phosphorylation; Protective Agents; Protein Serine-Threonine Kinases; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Signal Transduction; Up-Regulation

Hepatic ischemia/reperfusion (I/R) injury is a major cause of high morbidity and mortality after liver resection, transplantation, and hemorrhagic shock. Paeoniflorin (PF), the main substance of glucosides in Radix Paeoniae Alba, has been widely used to treat various hepatic inflammatory diseases including I/R injury. However, the underlying mechanisms of PF on hepatic I/R injury remain further investigated. In this study, the liver I/R model was performed by clamping the portal vein and hepatic artery with an atraumatic clamp for 90 min followed by 6 hr reperfusion. PF (100 mg/kg) was given three times a day by gavage before I/R. The blood and hepatic samples were collected to evaluate liver injury and molecular indexes. The results showed that PF pretreatment significantly inhibited I/R-induced serum ALT and AST activities (40.3% and 53.8% those of I/R group, respectively), hepatic pathological damages and hepatic apoptosis (P < 0.01), and infiltration of neutrophils into liver. In addition, PF suppressed the production of pro-inflammatory cytokines (P < 0.01), decreased the expression of high mobility group box-1 (HMGB1), and down-regulated toll-like receptors 4 (TLR4) and phosphorylated ERK1/2, JNK1/2, p38, and NF-κB signal molecules expression in the I/R-operated mice. These findings indicated that PF played a protective role in liver I/R injury, and this protection was associated with inhibition of I/R-activated HMGB1-TLR4 signaling pathway to attenuate hepatic inflammation responses.

Topics: Animals; Apoptosis; Caspase 3; Down-Regulation; Glucosides; HMGB1 Protein; Interleukin-1beta; Liver; Male; Mice; Mice, Inbred C57BL; Monoterpenes; NF-kappa B; Paeonia; Protective Agents; Reperfusion Injury; Signal Transduction; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

In this study, in-depth observation and investigation of blood-brain barrier permeability and neuroprotective effect of Trichosanthes kirilowii cassia twig particles on rats with cerebral ischemia-reperfusion injury were performed. Focal cerebral ischemia-reperfusion injury model was established by middle cerebral artery occlusion method, reperfusion was implemented 2 hours after ischemia; qualitative analysis and investigation of trichosanthes kirilowii cassia twig particles in plasma, brain tissue and cerebrospinal fluid in normal and middle cerebral artery occlusion (MCAO) rats were done by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS); changes in neurological deficits, cerebral infarction stereometry, blood-brain barrier permeability and histopathological changes of MCAO model rats were observed. Qualitative analysis by HPLC-MS/MS results showed that ingredients, paeoniflorin, albiflorin, liquiritin in Trichosanthes kirilowii cassia twig particles can reach the brain through the blood-brain barrier. In the model group, glycyrrhizin and glycyrrhizic acid can be detected in brain tissue or cerebrospinal fluid. In addition, Trichosanthes kirilowii cassia twig particles can significantly lower neurological deficits of rats in middle cerebral artery occlusion model, reduce the Evans blue penetration, contract infarct size, and reduce pathological tissue injury of cerebral ischemia reperfusion. The ingredients of Trichosanthes kirilowii cassia twig particles can reach the brain tissue through the blood-brain barrier and play a role in neuroprotection of rats with cerebral ischemia-reperfusion injury, which has important research significance and brings scientific experimental, theoretical basis for clinical drug use.

Topics: Animals; Blood-Brain Barrier; Brain; Bridged-Ring Compounds; Flavanones; Glucosides; Glycyrrhizic Acid; Infarction, Middle Cerebral Artery; Male; Monoterpenes; Neuroprotective Agents; Plant Components, Aerial; Plant Extracts; Rats; Recovery of Function; Reperfusion Injury; Trichosanthes

Xin-Shao formula is a folk remedy widely used in China to prevent and cure stroke. Cerebral ischemic reperfusion (I/R) injury often takes place during the treatment of stroke. Information about the pharmacokinetic behavior of the remedy under cerebral I/R injury conditions is lacking. The present study aimed to compare the pharmacokinetic properties of scutellarin and paeoniflorin, two major bioactive components of Xin-Shao formula, under physiological state in cerebral I/R injury rats. Neurobehavioral dysfunction was evaluated and cerebral infarcted volume was measured in middle cerebral artery occlusion I/R injury (MCAO) rats. Plasma samples were collected at various time points after a single dose (intravenous, i.v.) of Xin-Shao formula. The levels of plasma scutellarin and paeoniflorin at the designed time points were determined by a UPLC-MS/MS method, and drug concentration versus time plots were constructed to estimate pharmacokinetic parameters. Increase in terminal elimination half-life (t1/2z) and mean residence time (MRT(0-t)) of scutellarin as well as elevation in area under the plasma drug concentration-time curve from 0 h to the terminal time point (AUC(0-t)) and maximum plasma drug concentration (Cmax) of paeoniflorin, along with decreased clearance of paeoniflorin and scutellarin as well as reduced apparent volume of distribution (Vz) of paeoniflorin, were observed in MCAO rats, compared with those in sham-operated animals. The elimination of scutellarin and paeoniflorin were reduced in cerebral I/R injury reduced rats.

Topics: Animals; Apigenin; Disease Models, Animal; Glucosides; Glucuronates; Infarction, Middle Cerebral Artery; Male; Medicine, Chinese Traditional; Monoterpenes; Plant Extracts; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Stroke

This study is to investigate the effects of paeoniflorin on cerebral blood flow and the balance of PGI2/TXA2 of rats with focal cerebral ischemia-reperfusion injury. A total of 72 SD rats (3) were randomly divided into 6 groups: sham operation group, cerebral ischemia-reperfusion model group (I/R gourp), low (10 mg.kg-1), middle (20 mg.kg-1) and high (40 mg.kg-1) doses of paeoniflorin groups and nimrnodipine group. Focal cerebral ischemia in rats was made by inserting a monofilament suture into internal carotid artery for 90 min and then reperfused for 24 h. The effects of paeoniflorin on neurological deficit scores and the infarction volume of brain were detected. Relative regional cerebral blood flow (rCBF) was continuously monitored over ischemic hemispheres by laser-Doppler flowmetry (LDF). The expression of COX-2 in hippocampal CAl region was estimated by immunohistochemistry and the contents of prostacyclin I2 (PGI2), thromboxane A2 (TXA2), and ratio of PGIJ2/TXA2 in serum were measured by ELISA kits. Paeoniflorin significantly ameliorated neurological scores, reduced the infarction volume, and increased regional cerebral blood flow relative to the I/R group. In addition, paeoniflorin could inhibit COX-2 expression and the release of TXA2 and prevent the downregulation of PGI2 induced by I/R injury. The neuroprotective effects of paeoniflorin against focal cerebral ischemia-reperfusion rats might be attributed to improve the supply of injured hemisphere blood flow and adjust the balance between PGI2/TXA2.

Topics: 6-Ketoprostaglandin F1 alpha; Animals; Brain; CA1 Region, Hippocampal; Cyclooxygenase 2; Glucosides; Infarction, Middle Cerebral Artery; Male; Monoterpenes; Neuroprotective Agents; Paeonia; Plants, Medicinal; Random Allocation; Rats; Rats, Sprague-Dawley; Regional Blood Flow; Reperfusion Injury; Thromboxane B2

Paeoniflorin, a component in Paeonia lactiflora Pall, inhibits nuclear factor-kappaB expression in chronic hypoperfusion rat and has anti-inflammatory properties. Therefore, the aim of the present study was to investigate the effect of paeoniflorin on cerebral infarct, and the involvement of anti-inflammation. We established an animal model of cerebral infarct by occluding both the common carotid arteries and the right middle cerebral artery for 90 min, followed by reperfusion of 24 hours. The ratios of cerebral infarction area to total brain area, and neuro-deficit score were used as an index to observe the effects of paeoniflorin on cerebral infarct. ED1 (mouse anti rat CD68), interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), intercellular adhesion molecular-1 (ICAM-1), myeloperoxidase (MPO) immunostaining and apoptotic cells in the cerebral infarction region also were studied. The results indicated that both pre-treatment and post-treatment with paeoniflorin reduced the ratio of cerebral infarction area; pre-treatment with paeoniflorin also reduced the neurological deficit score. The counts of ED1, IL-1beta, TNF-alpha, ICAM-1 of microvessels and MPO immunoreactive cells and apoptotic cells were increased in the cerebral infarction region; however, these increases were reduced by Paeoniflorin pre-treatment. In conclusion, Paeoniflorin reduced cerebral infarct and neurological deficit in ischemia-reperfusion injured rats, suggesting that paeoniflorin may have a similar effect in humans and might be a suitable treatment for stroke. Paeoniflorin reduced cerebral infarct, at least in part, involves the anti-inflammatory properties.

Topics: Animals; Anti-Inflammatory Agents; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Apoptosis; Benzoates; Brain; Bridged-Ring Compounds; Cerebral Infarction; Disease Models, Animal; Glucosides; Intercellular Adhesion Molecule-1; Interleukin-1beta; Monoterpenes; Nervous System Diseases; Paeonia; Peroxidase; Phytotherapy; Plant Extracts; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Tumor Necrosis Factor-alpha

The time course of paeoniflorin in the cortex of normal and cerebral ischemia-reperfusion rats, following intravenous administration of Paeoniae Radix extract at a dose of 60 mg/kg of paeoniflorin, was determined using high-performance liquid chromatographic (HPLC) assay. The results showed that paeoniflorin could penetrate through the blood-brain barrier to reach the cortex, and that the injuries of ischemia-reperfusion could play an important role in pharmacokinetic process of paeoniflorin in the cortex after intravenous administration of Paeoniae Radix extract. The cortex concentrations of paeoniflorin in cerebral ischemia-reperfusion rats were lower 5 min after dosing and declined more slowly than that in normal control.

Topics: Animals; Benzoates; Brain Ischemia; Bridged-Ring Compounds; Cerebral Cortex; Chromatography, High Pressure Liquid; Glucosides; Male; Monoterpenes; Paeonia; Plant Extracts; Rats; Rats, Wistar; Reperfusion Injury; Reproducibility of Results

To explore the effects of paeoniflorin on antagonising the delayed neuronal death (DND) induced by cerebral ischemia,and the relation between DND, cerebral tissue energy metabolism, nitric oxide (NO) and nitric oxide synthase (NOS).. Incomplete cerebral ischemia induced was induced by ligating bilateral arteries carotis communis for 20 min followed by reperfusion 48 h in rats. The indexes including Na(+)-K(+)-ATPase activity, lactic acid content, Ca(2+)-ATPase, Mg(2+)-ATPase activity, NO content and NOS activity were determined in fore brain cortex at 48 h after reperfusion.. Na(+)-K(+)-ATPase, Ca(2+)-ATPase and Mg(2+)-ATPase activity were lowered (P < 0.01), NO level was decreased (P < 0.01), NOS activity dropped (P < 0.01) in cerebral tissue at 48h after reperfusion, but lactic acid level had no change. Paeoniflorin could prevent reduction of Na(+)-K(+)-ATPase activity (P < 0.05, P < 0.01), increase NO level (P < 0.01), enhance NOS activity (P < 0.01) at 48h after reperfusion.. DND induced by ischemia may be concerned with energy metabolism disorder and decrease of NO formation. Paeoniflorin may play the role of antagonising cerebral ischemia by adjusting cerebral energy metabolism and nitric oxide formation.

Topics: Animals; Benzoates; Brain; Brain Ischemia; Bridged-Ring Compounds; Ca(2+) Mg(2+)-ATPase; Calcium-Transporting ATPases; Energy Metabolism; Female; Gerbillinae; Glucosides; Lactic Acid; Male; Monoterpenes; Nitric Oxide; Nitric Oxide Synthase; Paeonia; Plants, Medicinal; Reperfusion Injury; Sodium-Potassium-Exchanging ATPase

The objective of the present study was to investigate the effects of cerebral ischemia-reperfusion on pharmacokinetics of paeoniflorin after intravenous administration of Paeoniae Radix extract (PRE) in rats. The cerebral ischemia-reperfusion rats were induced by occluding the bilateral carotid arteries of normal rats for 2 h, followed by reperfusion. The resultant animals were immediately administrated by PRE (at a dose of 60 mg/kg of paeoniflorin) via the femoral vein, whilst the same dose was injected to the normal rats. Plasma samples were collected at different time to construct pharmacokinetic profiles by plotting drug concentration versus time. Quantification of paeoniflorin in rat plasma was achieved by using a simple and rapid high-performance liquid chromatographic method. In normal rats, the major parameters of distribution half-life (t1/2alpha), elimination half-life (t1/2beta), area under the plasma concentration-time (AUC), mean retention time (MRT), and clearance (CL), estimated by an open two-compartmental model, were 0.69, 18.77 min, 5338.71 (microg min)/ml, 18.13 min and 0.0162 mg/(kg min), respectively. However, in ischemia-reperfusion rats, the corresponding parameters were 2.04, 24.51 min, 9626.00 (microg min)/ml, 29.75 min and 0.0071 mg/(kg min), respectively. The results showed that ischemia- reperfusion significantly increased AUC values, decreased CL values, and prolonged the terminal half-life of paeoniflorin. These findings suggest that the injuries of ischemia-reperfusion could play an important role in pharmacokinetic process of paeoniflorin.

Topics: Animals; Benzoates; Brain Ischemia; Bridged-Ring Compounds; Glucosides; Infusions, Intravenous; Male; Monoterpenes; Paeonia; Plant Extracts; Rats; Rats, Wistar; Reperfusion Injury