crocin and Reperfusion-Injury

Topics: Animals; Antioxidants; Carotenoids; Crocus; Free Radical Scavengers; Humans; Plant Extracts; Rats; Reperfusion Injury

As a powerful antioxidant compound, crocin can partially protect against renal ischemia/reperfusion (I/R) injuries. The encapsulation of components in niosomes (non-ionic surfactant-based vesicle) as nano-sized carrier systems has been proposed as they improve the solubility, stability, and bioavailability of drugs. Herein, the encapsulation of crocin in nano-niosomes and the effects of crocin-loaded nano-niosomes on renal ischemia/reperfusion-induced damages were evaluated. Nano-niosomes containing crocin were formulated by a modified heating method and were characterized for their physicochemical characteristics. Ischemia was induced by clamping the renal artery for 30 min followed by 1 or 24 h of reperfusion. Rats received an intra-arterial injection of nano-niosome-loaded crocin at the outset of reperfusion. Blood samples were taken after reperfusion to measure urea, creatinine (Cr), malondialdehyde (MDA), and superoxide dismutase (SOD) activity. The right kidney was removed for histological examination. The results showed that crocin-contain nano-niosomes have appropriate size and morphology, acceptable encapsulation efficiency, and a proper release pattern of crocin. I/R enhanced creatinine (Cr), urea, and malondialdehyde (MDA) serum levels and reduced SOD activity and histological damages in the renal tissue.

Topics: Animals; Antioxidants; Blood Urea Nitrogen; Carotenoids; Creatinine; Glutathione Peroxidase; Kidney; Kidney Diseases; Liposomes; Male; Malondialdehyde; Nanoparticles; Oxidative Stress; Protective Agents; Rats; Rats, Wistar; Renal Artery; Reperfusion Injury; Superoxide Dismutase

Oxidative stress, caused by extreme accumulation of un-scavenged reactive oxygen species, plays an integral role in the Ischemia-Reperfusion (I/R) injury to the testicles following testicular torsion. The current research aimed to examine the protective effects of crocin as a natural antioxidant on testicular I/R injury in rats. Animals were divided randomly into five groups (seven each): (1) sham group, (2) torsion/detorsion (T/D) group, (3) intact group with 100 mg/kg crocin, (4) and (5) T/D groups followed by treatment with two different doses of crocin (50 and 100 mg/kg (IP)). I/R injury was induced by 720° clockwise torsion of the left testicles for 2 h. After 24 h of reperfusion, blood samples and epididymal sperms were collected to measure biochemical (GPx, SOD, and MDA), hormonal (testosterone), and sperm parameters (total sperm recovery, motility, viability, and morphology). Moreover, affected testicles were subjected to histopathology examination. I/R injury caused a significant reduction in sperm characteristics (except for morphology) (P < 0.05), which could not be significantly improved by crocin administration at either dose (P > 0.05). Johnsen's testicular score, mean seminiferous tubular diameter, and germinal epithelial cell thickness were significantly decreased in the T/D group compared to the intact and sham groups. However, crocin could significantly improve the histopathological parameters in both treatment groups compared to the T/D group (P < 0.05). T/D reduced SOD and GPx activity and testosterone level significantly (except for GPx) compared to the sham group (P < 0.05). However, crocin administration could significantly reverse them. Also, crocin reduced the amount of MDA significantly in the high-dose treatment group in comparison to T/D group (P < 0.05). The results of the current study revealed that crocin could be a promising agent to protect against I/R injury following surgical correction of the testicular torsion.

Topics: Animals; Carotenoids; Male; Malondialdehyde; Rats; Reperfusion Injury; Rodent Diseases; Spermatic Cord Torsion; Testis

Renal Ischemia (RI) usually develops as a secondary manifestation of hypertension, various cardiovascular disorders and renal transplantation. It exerts hypoxic oxidative stress to kidneys, together with stimulation of several immune-mediated inflammatory cascades. Such events eventually damage renal tubules and glomeruli, driving acute kidney injury (AKI) and ultimately, renal failure. Crocin; the main bioactive constituent of Crocus sativus extract has been reported to demonstrate numerous pharmacological merits. In the current study, unilateral renal ischemia reperfusion injury (URIRI) was induced in rats by unilateral clamping of the left renal pedicle for 45 min followed by 24 h of reperfusion. Daily pre-treatment with crocin (20 mg/kg, orally) for 7 days, significantly improved all signs of renal injury. Biochemically, kidney functions; including serum creatinine (Sr Cr), blood urea nitrogen (BUN), proteinuria and creatinine clearance (Cr Cl) significantly improved. Inflammatory biomarkers; serum lactate dehydrogenase (LDH) and kidney nitric oxide (Nos) contents significantly declined. Oxidant/antioxidant balance was significantly restored; manifested in recovery of renal superoxide dismutase (SOD) activity, glutathione (GSH) concentration, malondialdehyde (MDA) content and restoration of serum catalase activity. Kidney contents of inflammatory cytokine interleukin-6 (IL6) and toll-like receptors 4 (TLR4) significantly declined as well. Histopathologically, crocin pretreatment resulted in signs of improvement with minimal renal lesions with significant decrease in renal inflammatory cells count. In conclusion, crocin induced restoration of normal kidney functions is mediated through multiple mechanisms including mainly attenuation of oxidative stress and inflammation via down-regulation of renal TLR4 and IL6 expression.

Topics: Animals; Anti-Inflammatory Agents; Antioxidants; Blood Urea Nitrogen; Carotenoids; Catalase; Glutathione; Interleukin-6; Kidney; Kidney Diseases; L-Lactate Dehydrogenase; Male; Malondialdehyde; Nitric Oxide; Oxidative Stress; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Superoxide Dismutase; Toll-Like Receptor 4

In this study, the antioxidant activity of crocin via experimental and theoretical methods was investigated. In order to induce oxidative stress, 30-min renal ischemia and 24-h reperfusion were used in male Wistar rats. Oxidative stress was assessed by measuring tissue malondialdehyde (MDA) and ferric reducing/antioxidant power (FRAP). The results showed that following ischemia/reperfusion, the level of MDA was increased and FRAP decreased. Both of these changes were alleviated by crocin administration. The bond dissociation enthalpy and ionization potential values as enthalpies of mechanism of antioxidant activity of crocin were calculated by density functional theory method. According to obtained results, the novel structures of crocin with higher antioxidant activity for synthesis were proposed. Results indicated that NH

Topics: Animals; Antioxidants; Carotenoids; Drug Delivery Systems; Male; Models, Theoretical; Nanotubes; Oxidative Stress; Rats; Rats, Wistar; Reperfusion Injury; Structure-Activity Relationship

The aim of this study was to investigate the comparative protective effects of separate and combined pretreatment with Cr and ZnSO4 on serum levels of miR-122, miR-34a, liver function tests, protein expression of Nrf2 and p53, and histopathological changes following IR-induced hepatic injury.. Fifty-six male Wistar rats randomly assigned into seven groups (n=8). Sham (S), IR, crocin pretreatment (Cr), and crocin pretreatment+IR (Cr+IR), ZnSO4 pretreatment (ZnSO4), ZnSO4 pretreatment+IR (ZnSO4+IR) and their combination (Cr+ZnSO4+IR) groups. In sham, ZnSO4 and Cr groups, animals received normal saline (N/S, 2ml/day), Cr (200mg/kg) and ZnSO4 (5mg/kg) for 7 consecutive days (intraperitoneally; i.p), then only laparotomy was performed. In IR, Cr+IR, ZnSO4+IR and Cr+ZnSO4+IR groups, rats received N/S, Cr and ZnSO4 with same dose and time, then underwent a partial (70%) ischemia for 45min that followed by reperfusion for 60min. Blood sample was taken for biochemical and microRNAs assay, tissue specimens were obtained for antioxidants, protein expression, histopathological and immunohistochemical evaluations.. The results showed that Cr and ZnSO4 increased antioxidants activity and expression of Nrf2, decreased serum levels of liver enzymes, miR-122, miR-34a, p53 expression and also ameliorated histopathological abnormality. However, their combination caused more improvement on IR-induced liver injury.. This study demonstrated that Cr, ZnSO4 and their combination through increasing antioxidant activity and Nrf2 expression, decreasing the serum levels of liver enzymes, miR-122, 34a, p53 expression, and amelioration of histopathological changes, protected liver against IR-induced injury.

Topics: Animals; Carotenoids; Drug Therapy, Combination; Liver; Male; MicroRNAs; NF-E2-Related Factor 2; Rats; Rats, Wistar; Reperfusion Injury; Zinc Sulfate

Cerebral ischemia reperfusion (IR) injury is a process in which oxidative and apoptotic mechanisms play a part. Neuroprotective agents to be found could work out well for the efficient and safe minimization of cerebral IR injury. Crocin is a strong antioxidant agent; however the influence of this agent on the experimental cerebral ischemia model has not been studied extensively and thus it is not well-known. The objective of our study was to investigate the antioxidant, antiapoptotic and protective effects of crocin on the global cerebral IR induced by four-vessel occlusion.. A total of 30 adult female Sprague-Dawley rats were equally and randomly separated into three groups as follows: sham, IR and IR+crocin (40mg/kg/day orally for 10days). 24h after electrocauterization of bilateral vertebral arteries, bilateral common carotid arteries were occluded for 30min and reperfused for 30min. Oxidative stress parameters (TAS, TOS, OSI), haematoxylin and eosin staining, caspase-3 and hypoxia-inducible factor-1 alpha (HIF-1α) expressions and TUNEL methods were investigated.. There was a significant difference between the IR and sham groups by means of OSI level, histopathological scoring, caspase-3, HIF-1α and TUNEL-positive cell parameters. We have also observed that pre-treatment with crocin reduced these parameter levels back to the baseline.. The data obtained from the present study suggest that crocin may exert antiapoptotic, antioxidant and protective effects in IR-mediated brain injury induced by four-vessel occlusion. To the best of our knowledge, this would be the first study to be conducted in this field.

Topics: Animals; Antioxidants; Apoptosis; Brain Ischemia; Carotenoids; Cerebrovascular Circulation; Female; Rats; Rats, Sprague-Dawley; Reperfusion Injury

Ischemia and reperfusion (IR) injury is an important complication of abdominal aortic surgery, and it mainly affects the lower extremities and remote organs. In the present study, we aimed to investigate the possible protective effect of crocin in IR-mediated kidney damage.. A total of 24 adult male Wistar-Albino rats were equally and randomly separated into three groups as follows: sham laparotomy, IR, and IR + crocin. Infrarenal aortic occlusion and reperfusion was applied for 1 and 2 h, respectively. Tissue samples were removed and collected. Biochemical and histopathologic analyses were performed.. Urea, blood urea nitrogen, creatinine, renal tissue tumor necrosis factor α, interleukin (IL)-6, IL-18, interferon gamma, IL-1β, total oxidant status, and oxidative stress index levels were significantly higher in IR group, when compared with other groups. These improvements were also demonstrated with some parameters including total score of histopathologic damage, Tunel, Bax, and Caspase-3 expression levels, and these parameters were prominently higher in the IR group, when compared with the other groups. Nevertheless, Bcl2 expression degree was prominently lower in the IR group than those in the other two groups.. Data established from the present study suggest that crocin can preclude renal damage in infrarenal aortic occlusion models.

Topics: Acute Kidney Injury; Animals; Aorta, Abdominal; Biomarkers; Carotenoids; Drug Administration Schedule; Male; Protective Agents; Random Allocation; Rats; Rats, Wistar; Reperfusion Injury; Treatment Outcome

Ischemia-reperfusion (IR) injury of the liver is an unresolved problem that occurs during certain surgical approaches, including hepatic, cardiac and aortic operations. In this study we aimed to investigate whether crocin and safranal had protective effects on liver IR injury induced in an infrarenal aortic clamping (IRAC) model. Male Wistar-Albino rats (n=32) were divided into four groups with 8 animals each as follows: Sham, IR, IR+crocin, and IR+safranal. The infrarenal aorta (IRA) was clamped for 60min for the ischemic period and allowed to reperfuse for 120min. Blood and tissue samples were collected for biochemical, histological and immunohistological analysis. Plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were found to be significantly higher in the IR group than the sham group (respectively; p=0.015, p<0.001). There were significant differences between the IR group and the IR+crocin group or the IR+safranal group in AST levels (respectively; p=0.02, p<0.001). ALT showed a significant decrease in the IR+crocin group compared to the IR group (p<0.05). We also observed histopathological changes among the groups. Bax and Caspase-3 expression in the IR group was remarkably higher than in the other groups. Caspase-3 and Bax expression in the IR+crocin and the IR+safranal groups were significantly lower than in the IR group. Nevertheless, there were no significant differences in BCL2 expression among the groups. IRAC is a cause of IR injury in the liver. This study showed that crocin and safranal have protective effects on IR induced liver injury.

Topics: Alanine Transaminase; Animals; Arterial Occlusive Diseases; Aspartate Aminotransferases; bcl-2-Associated X Protein; Carotenoids; Caspase 3; Cyclohexenes; Immunohistochemistry; Kidney; Liver; Liver Function Tests; Male; Rats, Wistar; Reperfusion Injury; Terpenes

Crocin, a pharmacologically active component of Crocus sativus L. (saffron), has been reported to be useful in the treatment of oxidative stress injury. In the present study, we investigated the antioxidative effect of crocin and the change of the ERK signaling pathway on rat retina induced by ischemia/reperfusion (IR) injury.. Crocin was pretreated 30 min before and once daily after retinal IR injury by intraperitoneal injection. The retinal morphological damage was observed by hematoxylin and eosin (HE) staining. The number of retinal ganglion cells (RGCs) was counted by Brn-3a immunofluorescence staining. The antiapoptotic effect of crocin was determined by detecting cleaved caspase-3 protein levels by means of Western blot and immunohistochemical analysis. Furthermore, retinas were also used for the determination of malondialdehyde (MDA) levels, glutathione (GSH) levels, total superoxide dismutase (T-SOD), and reactive oxygen species (ROS). The phosphorylated ERK (p-ERK) protein level was determined by Western blot and immunohistochemical analysis.. Our results showed that crocin treatment (50 mg/kg) significantly inhibited the decrease of retinal thickness through HE staining and protected RGCs from decreasing. Compared with the IR + vehicle group, crocin treatment significantly decreased cleaved caspase-3 and p-ERK protein expression by Western blot analysis and immunohistochemistry. Immunohistochemical staining for cleaved caspase-3 and p-ERK in the retinal sections showed positive cells were present in the RGC layer and inner nuclear layer after IR injury. Similarly, crocin (50 mg/kg) treatment also significantly increased the level of activity of GSH, enhanced the activity of T-SOD, and decreased the activity level of ROS and MDA after IR injury.. These findings demonstrated that crocin treatment could notably protect the retina from damage induced by IR. It might be related to crocin antioxidant and antiapoptotic properties in the retina.

Topics: Animals; Apoptosis; Blotting, Western; Carotenoids; Caspase 3; Cell Survival; Free Radical Scavengers; Glutathione; Immunohistochemistry; Male; Malondialdehyde; MAP Kinase Signaling System; Neuroprotective Agents; Oxidative Stress; Rats; Rats, Sprague-Dawley; Reactive Oxygen Species; Reperfusion Injury; Retinal Diseases; Retinal Ganglion Cells; Superoxide Dismutase

Crocin is a pharmacologically active component of Crocus sativus L. (saffron) and has been reported to be useful in the treatment of neuronal damage. In the present study, we investigated the neuroprotective effect of crocin on retinal ganglion cells (RGCs) after retinal ischaemia/reperfusion (IR) injury, and our results show that crocin acts through the PI3K/AKT signalling pathway. Retinal IR injury was induced by raising the intraocular pressure of Sprague-Dawley rats to 110 mmHg for 60 min. The neuroprotective effect of crocin was determined by quantifying the surviving RGCs and apoptotic RGCs following IR injury by means of retrograde labelling and TUNEL staining, respectively. The phosphorylated AKT protein level was determined by western blot and immunohistochemical analysis. To determine the extent to which the PI3K/AKT pathway contributes to the neuroprotective effect of crocin, experiments were also performed using the PI3K inhibitor LY294002. Compared with the IR + vehicle group, crocin (50 mg/kg) treatment enhanced RGC survival by approximately 36% and decreased RGC apoptosis by 44% after retinal IR injury. Western blot and immunohistochemical analysis demonstrated that the PI3K/AKT pathway was activated by crocin in the ganglion cell layer after retinal IR injury. Intravitreal injection of LY294002 blocked the neuroprotective effect of crocin on IR-induced RGC death. In conclusion, crocin prevents retinal IR-induced apoptosis of RGCs by activating the PI3K/AKT signalling pathway.

Topics: Animals; Apoptosis; bcl-2-Associated X Protein; Blotting, Western; Carotenoids; Cell Survival; Chromones; Enzyme Inhibitors; In Situ Nick-End Labeling; Male; Morpholines; Neuroprotective Agents; Phosphatidylinositol 3-Kinase; Phosphorylation; Proto-Oncogene Proteins c-akt; Proto-Oncogene Proteins c-bcl-2; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Retinal Ganglion Cells; Signal Transduction

This paper studied the effects of crocin, a pharmacologically active component of Crocus sativus L., on ischemia/reperfusion (I/R) injury in mice cerebral microvessels. Transient global cerebral ischemia (20 min), followed by 24 h of reperfusion, significantly promoted the generation of nitric oxide (NO) and malondialdehyde (MDA) in cortical microvascular homogenates, as well as markedly reduced the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) and promoted the activity of nitric oxide synthase (NOs). Reperfusion for 24 h led to serous edema with substantial microvilli loss, vacuolation, membrane damage and mitochondrial injuries in cortical microvascular endothelial cells (CMEC). Furthermore, enhanced phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and decreased expression of matrix metalloproteinase-9 (MMP-9) were detected in cortical microvessels after I (20 min)/R (24 h). Reperfusion for 24 h also induced membrane (functional) G protein-coupled receptor kinase 2 (GRK2) expression, while it reduced cytosol GRK2 expression. Pretreatment with crocin markedly inhibited oxidizing reactions and modulated the ultrastructure of CMEC in mice with 20 min of bilateral common carotid artery occlusion (BCCAO) followed by 24 h of reperfusion in vivo. Furthermore, crocin inhibited GRK2 translocation from the cytosol to the membrane and reduced ERK1/2 phosphorylation and MMP-9 expression in cortical microvessels. We propose that crocin protects the brain against excessive oxidative stress and constitutes a potential therapeutic candidate in transient global cerebral ischemia.

Topics: Animals; beta-Adrenergic Receptor Kinases; Blood Vessels; Brain Edema; Brain Ischemia; Carotenoids; Cerebral Cortex; Endothelial Cells; Extracellular Signal-Regulated MAP Kinases; G-Protein-Coupled Receptor Kinase 2; Glutathione Peroxidase; Male; Malondialdehyde; Matrix Metalloproteinase 9; Mice; Mice, Inbred C57BL; Microcirculation; Nitric Oxide; Nitric Oxide Synthase; Oxidative Stress; Phosphorylation; Reperfusion Injury; Subcellular Fractions; Superoxide Dismutase; Tissue Distribution

The generation of reactive oxygen species and lipid peroxidation are associated with tissue injury following ischemic insult; therefore, the use of antioxidants appears rational in the improvement of kidney diseases therapy. The aim of the present study was to assess the effect of aqueous saffron extract (Crocus sativus L.) and its active constituent, crocin, on oxidative stress following renal ischemia-reperfusion injury (IRI) in rats.. The cellular redox status (thiobarbituric acid reactive species (TBARS) and total thiol levels) and antioxidant power (using ferric reducing/antioxidant power test) were assessed in control and ischemic groups. The left kidney was exposed to warm ischemia for 60 min followed by reperfusion for 90 min. The macerated aqueous extract of saffron (with doses of 5, 20 and 80 mg/kg, i.p.) and crocin (with doses of 50, 200 and 400 mg/kg, i.p.) were administrated prior to induction of ischemia. Normal saline (10 ml/kg, i.p.) was injected to control group and a sham group that did not have ischemia-reperfusion.. Ischemia-reperfusion (IR) caused a significant increase in TBARS levels (p<0.001) and decrement in both antioxidant power (FRAP value) (p<0.05) and total thiol concentration (p<0.001) in kidney homogenate samples. In crocin pretreated groups, a reduction in TBARS levels (from 85.8 +/- 5.4 to 20.9 +/- 1.5 nmol/g tissue, p<0.001; 400 mg/kg) and elevation in antioxidant power (FRAP value) (from 3.05 +/- 0.16 to 4.15 +/- 0.16 micromol/g tissue, p<0.001; 400 mg/kg) and total thiol concentrations (from 0.38 +/- 0.03 to 0.62 +/- 0.03 mM, p<0.001; 200 mg/kg), as compared with control group, were observed. The aqueous extract also reduced lipid peroxidation products (from 85.8 +/- 5.4 to 15.9 +/- 2.6 nmol/g tissue, p<0.001; 80 mg/kg) and increased antioxidant power (from 2.98 +/- 0.11 to 5.97 +/- 0.56 micromol/g tissue, p<0.001; 80 mg/kg) in ischemia-reperfusion injured rat kidneys.. This study therefore suggests that the aqueous saffron extract (Crocus sativus L.) and its active constituent, crocin, may be useful agents for the prevention of renal ischemia-reperfusion (IR)-induced oxidative injury in rats.

Topics: Animals; Carotenoids; Crocus; Dose-Response Relationship, Drug; Flowers; Kidney; Male; Oxidative Stress; Plant Extracts; Rats; Rats, Wistar; Reperfusion Injury