trans-sodium-crocetinate and Reperfusion-Injury

Uterine ischemia/reperfusion (I/R) injury often occurs during many complex surgical procedures, such as uterus transplantation, cesarean, and myomectomy, which may lead to the loss of uterine function and failure of the operation. Crocetin (CRO), as one of the major active constituents from saffron extract, shows protective effects against reactive oxygen species, inflammation, and apoptosis. However, the role of CRO in protecting the uterus against I/R-induced injury has never been investigated. This study aims to clarify the protective role of CRO against I/R injury and the underlying mechanisms.. Sprague-Dawley rats were randomly divided into five groups: the control group, I/R group, 20 mg/kg CRO-treated I/R group, 40 mg/kg CRO-treated I/R group, and 80 mg/kg CRO-treated I/R group. Rats were given daily gavages with different doses of CRO or vehicle for five consecutive days. The rat uterine I/R model was created by routine method with 1h ischemia and 3h reperfusion. The serum and uterine tissues were collected, the changes in malondialdehyde (MDA) level and superoxide dismutase (SOD) activity, the mRNA and protein levels of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α and IL-10, the protein levels of B-cell chronic lymphocytic leukemia/lymphoma (Bcl)-2, Bcl-2-associated X protein (Bax), caspase-3, nuclear factor erythroid 2-related factor (Nrf)-2, and heme oxygenase (HO)-1, were measured. The histological changes were examined by HE staining. The number of apoptotic cells was analyzed by flow cytometry.. Uterine I/R significantly induced MDA level, suppressed SOD activity, upregulated levels of pro-inflammatory cytokines, down-regulated level of the antiinflammatory cytokine, induced caspase-3-dependent apoptosis, activated the protein expression of Nrf-2 and HO-1, and caused uterine damage. However, pre-administration of CRO effectively reversed I/R-induced above changes and further enhanced Nrf-2/HO- 1 activation in a dose-dependent manner.. Pre-administration of CRO effectively alleviates I/R-induced oxidative stress, inflammation, apoptosis, and tissue injury probably through activating the Nrf- 2/HO-1 pathway, suggesting a protective role of CRO in I/R-induced uterus injury.

Topics: Animals; Apoptosis; Caspase 3; Cytokines; Female; Inflammation; Ischemia; NF-E2-Related Factor 2; Oxidative Stress; Rats; Rats, Sprague-Dawley; Reperfusion; Reperfusion Injury; Superoxide Dismutase; Tumor Necrosis Factor-alpha

Trans sodium crocetinate (TSC) has been reported to exert a protective effect against cerebral ischemia/reperfusion (I/R) injury. However, whether TSC protects against myocardial ischemia/reperfusion (MI/R) injury remains unknown. Herein, we found that TSC treatment reduced myocardial infract size and elevated serum LDH and CK activities of MI/R rats. TSC administration attenuated oxidative stress in MI/R rats and H9C2 cells exposed to oxygen glucose deprivation/reperfusion (OGD/R). TSC administration relieved I/R-induced myocardial apoptosis in vivo and in vitro, as evidenced by reduced number of TUNEL positive cells, accompanying with marked decreases in caspase-3 activity and Bax protein level and an increase in Bcl-2 protein level. TSC treatment markedly increased SIRT3 activity and SIRT3 and SOD2 protein levels, and could also diminished the phosphorylation of FOXO3a protein. Additionally, TSC treatment attenuated the acetylation of FOXO3a and SOD2 protein. But, these effects were obviously blocked by SIRT3 knockdown. Besides, SIRT3 knockdown blocked the cardioprotective effect of TSC on OGD/R-induced oxidative stress, apoptosis and mitochondrial dysfunction in vitro. In summary, TSC alleviates I/R-induced myocardial oxidative stress and apoptosis via the SIRT3/FOXO3a/SOD2 signaling pathway. Our study suggests that TSC may become a novel drug for the treatment of MI/R injury.

Topics: Animals; Antioxidants; Apoptosis; Carotenoids; Coronary Occlusion; Disease Models, Animal; Forkhead Box Protein O3; Humans; Male; Myocardial Infarction; Myocardium; Oxidative Stress; Rats; Rats, Wistar; Reperfusion Injury; RNA, Small Interfering; Signal Transduction; Sirtuin 3; Vitamin A

Crocetin, an aglycone of crocin, is found both in the saffron crocus (Crocus starus L.) and in gardenia fruit (Gardenia jasminoides Ellis). We evaluated the protective effects of crocetin against the retinal ischemia induced by 5h unilateral ligation of both the pterygopalatine artery (PPA) and the external carotid artery (ECA) in anesthetized mice. The effects of crocetin (20mg/kg, p.o.) on ischemia/reperfusion-induced retinal damage were examined by histological, electrophysiological, and anti-apoptotic analyses. Data for anti-apoptotic analysis was obtained by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Using immunohistochemistry and immunoblotting, the protective mechanism mediating the effects of crocetin was evaluated by examining crocetin's effects on the expression of 8-hydroxy-2-deoxyguanosine (8-OHdG; used as a marker of oxidative stress) and on phosphorylations of mitogen-activated protein kinases [MAPK; viz. extracellular signal-regulated protein kinases (ERK), c-Jun N-terminal kinases (JNK) and p38], and the redox-sensitive transcription factors nuclear factor-kappa B (NF-κB) and c-Jun. The histological analysis revealed that ischemia/reperfusion (I/R) decreased the cell number in the ganglion cell layer (GCL) and the thickness of inner nuclear layer (INL), and that crocetin inhibited GCL and INL. ERG measurements revealed that crocetin prevented the I/R-induced reductions in a- and b-wave amplitudes seen at 5 days after I/R. In addition, crocetin decreased the numbers of TUNEL-positive cells and 8-OHdG-positive cells, and the phosphorylation levels of p38, JNK, NF-κB, and c-Jun present in the retina after I/R. These findings indicate that crocetin prevented ischemia-induced retinal damage through its inhibition of oxidative stress.

Topics: Animals; Apoptosis; Carotenoids; DNA; Electroretinography; JNK Mitogen-Activated Protein Kinases; Male; Mice; Mitogen-Activated Protein Kinases; NF-kappa B; Oxidation-Reduction; Protective Agents; Reperfusion Injury; Retinal Diseases; Vitamin A