Ameliorative impacts of floral extract of Salvia species on oxidative stress and inflammation in rats renal ischemia/reperfusion

I/R injury is a potentially serious problem that is encountered during a variety of medical and surgical procedures, such as thrombolytic therapy, organ transplantation and coronary angioplasty, The basic pathophysiology of I/R injury is microvascular dysfunction which is developed following reperfusion of ischemic tissues. It has clinical importance because of its frequent occurrence and mortality in some surgical conditions such as renal transplantation. Here, we investigated the protective effect of Salvia extracts on kidneys against I/R injury. Forty Spraque Dawley rats were divided into 5 groups. Right nephrectomy was performed to all groups. Gr. I, control; Gr. II, I/R; Gr. III & IV, I/R+50 and I/R+100 mg/kg Salvia floral extract; and Gr. V with I/R+50 mg/kg Rosmarinic acid. Salvia and Rosmarinic acid for 7 days was given single dose as a gavage.60 min ischemia, 60 min reperfusion were applied to groups except control. Intracardiac blood samples were taken, Blood urea nitrogen, creatine, malondialdehyde, myeloperoxidase, nitric oxide and chitotriosidase levels were detected. Mean values were evaluated by statistical analysis. The renal tissues were examined under light microscopy. Based on our biochemical and histological data, Salvia floral extract has potent anti-inflammatory and antioxidant effects against renal structure and function.

A new combination of naringin and trimetazidine protect kidney Mitochondria dysfunction induced by renal Ischemia / Reperfusion injury in rat

Abstract Ischemia/reperfusion (IR) injury leads to overproduction of Reactive Oxygen Species (ROS), and disrupts membrane potential that contributes to cell death. The aim of this study was to determine if naringin (NAR), trimetazidine (TMZ) or their combination, protect the kidney mitochondrial from IR injury. Forty rats were randomly allocated into five groups, harboring eight rats each: Sham, IR, NAR (100 mg/kg), TMZ (5 mg/kg) and NAR plus TMZ. Ischemia was induced by obstructing both renal pedicles for 45 min, followed by reperfusion for 4 hours. The mitochondria were isolated to examine the ROS, Malondialdehyde (MDA), Glutathione (GSH), mitochondrial membrane potential (MMP) and mitochondrial viability (MTT). Our findings indicated that IR injury resulted in excessive ROS production, increased MDA levels and decreased GSH, MMP and MMT levels. However, NAR, TMZ or their combination reversed these changes. Interestingly, a higher protection was noted with the combination of both, compared to each drug alone. We speculate that this combination demonstrates a promising process for controlling renal failure, especially with the poor clinical outcome, acquired with NAR alone. This study revealed that pretreatment their combination serves as a promising compound against oxidative stress, leading to suppression of mitochondrial stress pathway and elevation of GSH level.

An in vivo study of neural protective effect of recombinant human KD/APP on rats with cerebral ischemia/reperfusion injury / 中国实验动物学报

Objective The amyloid β-protein precursor contains a domain highly homologous to Kunitz-type serine protease inhibitors. We have successfully established and characterized the recombinant human rhKD/APP in vitro. The aim of this study is to investigate the potential neuroprotective role of rhKD/APP on cerebral ischemia/reperfusion injury in rats. Methods Rats pretreated with rhKD/APP (4, 8, 16 mg/kg) were subjected to prepare models of cerebral ischemia/reperfusion (I/R) injury and those rats treated with Nimodipine were used as positive control. Comparison of the scores of neurological deficits, TTC-stained infarct volume and cerebral water content between the groups was performed. The activities of SOD, Na+-K+-ATPase and the content of MDA in the cortex tissues were measured and the activities of serum myeloperoxidase ( MPO) enzyme were also compared. The expressions of adhesion molecules ( ICAM-1 and E-selectin) were compared by immunohistochemistry. End-labeling of nuclear DNA fragmentation (TUNEL) and qualification of caspase-3, Bcl-2 and Bax were also employed to evaluate the local apoptosis in cortex tissues. Results By pretreatment with the rhKD/APP at three doses, cerebral infarct volume, water content and neurological deficits were all reduced. The activities of SOD, and Na+-K+-ATPase were increased, the contents of MDA were decreased in the cortex tissues, and the serum MPO activity was reduced. The expressions of adhesion molecules were downregulated and the apoptotic signaling of neurons were inhibited. All the changes induced by rhKD/APP treatment in the ischemia/reperfusion injury models showed statistical significance compared with the control rats. However, no significant difference was shown between the rhKD/APP group and Nimodipine group excepted for the reduced MPO in sera. Conclusions The result of this study suggest that rhKD/APP has neuroprotective effect on the cerebral ischemia/reperfusion injury through inhibiting multiple signaling pathways and is promising to be a potential neuroprotective drug.

Curcumin protects against the intestinal ischemia-reperfusion injury: involvement of the tight junction protein ZO-1 and TNF-alpha related mechanism

Present study aimed to investigate the eff ect of curcumin-pretreatment on intestinal I/R injury and on intestinal mucosa barrier. Thirty Wistar rats were randomly divided into: sham, I/R, and curcumin groups (n=10). Animals in curcumin group were pretreated with curcumin by gastric gavage (200 mg/kg) for 2 days before I/R. Small intestine tissues were prepared for Haematoxylin & Eosin (H&E) staining. Serum diamine oxidase (DAO) and tumor necrosis factor (TNF)-alpha levels were measured. Expression of intestinal TNF-alpha and tight junction protein (ZO-1) proteins was detected by Western blot and/or immunohistochemistry. Serum DAO level and serum and intestinal TNF-alpha leves were signifi cantly increased after I/R, and the values were markedly reduced by curcumin pretreatment although still higher than that of sham group (p<0.05 or p<0.001). H&E staining showed the significant injury to intestinal mucosa following I/R, and curcumin pretreatment signifi cantly improved the histological structure of intestinal mucosa. I/R insult also induced significantly down-regulated expression of ZO-1, and the eff ect was dramatically attenuated by curcumin-pretreatment. Curcumin may protect the intestine from I/R injury through restoration of the epithelial structure, promotion of the recovery of intestinal permeability, as well as enhancement of ZO-1 protein expression, and this eff ect may be partly attributed to the TNF-alpha related pathway.

Apoptosis signal transduction pathways in ischemia-reperfusion injury / 中国药学杂志

Ischemic-reperfusion (I/R) injury causes structural and functional damage to cells, eventually results in cells death. Apoptosis is known to be a vital mechanism that leads to cells death after ischemia followed by reperfusion. So the relationship between cell apoptosis and I/R injury has stimulated great interest in the development of anti-apoptotic therapies in brain I/R and myocardial I/R. However, there are many cell apoptosis signaling pathways which cross link with each other. This article makes a simple review between ischemic-reperfusion injury and cell apoptosis signal pathways.

Role of Nitric Oxide During Liver Transplantation.

Nitric oxide (NO) is a gaseous molecule produced from Nitric Oxide Synthases (NOS) enzyme. Three isoforms of NOS have been observed: endothelial NOS (eNOS), inducible NOS (iNOS) and neuronal NOS (nNOS). All three of these isoforms are expressed in liver in varying spatial and temporal ways. In liver, both nNOS and eNOS maintain homeostasis. Whereas iNOS is not expressed constitutively in liver, but rather is expressed in most liver cell types given the appropriate stimulatory conditions. Conflicting results have been observed on the behaviour and possible roles of the NO in several models of ischemia/ reperfusion injury during liver transplantation. Indeed, endogenous NO production has been associated with either protective or cytotoxic effects. Thus some, not all studies suggest that although eNOSderived NO production is protective in ischemia/reperfusion, iNOS derived NO production may contribute to ischemia/ reperfusion injury. This review article focuses on possible role of NO in liver transplantation.

Mediated protective effect of electroacupuncture pretreatment by miR-214 on myocardial ischemia/reperfusion injury / 老年心脏病学杂志（英文版）

Background Electroacupuncture pretreatment plays a protective role in myocardial ischemia/reperfusion (I/R) injury and microRNAs (miRNAs) could act on various facets of cardiac function. However, the role of miRNAs in the cardioprotection by electroacupuncture pre-treatment on myocardial I/R injury remains unknown. The purpose of the study was to examine whether miR-214 was involved in cardio-protection by electroacupuncture. Methods Using rat myocardial I/R model, we examined the role of electroacupuncture pretreatment in myocardial I/R injury and analyzed the changes in the expression of miR-214. In addition, I/R was simulated in vitro by performing oxy-gen-glucose deprivation (OGD) on H9c2 cell cultures, and the effect of electroacupuncture pretreatment on I/R injury as well as expressional level of miR-214 were examined in vitro. Furthermore, the miR-214 mimic was transfected into OGD-treated H9c2 cells, we analyzed the cell apoptosis, lactate dehydrogenase (LDH) and creatine kinase (CK) activities, intracellular free Ca2+concentration ([Ca2+]i) as well as the relative protein levels of sodium/calcium exchanger 1(NCX1), BCL2-like 11 (BIM), calmodulin-dependent protein kinase IIδ(CaMKIIδ) and Cyclophilin D (CypD). Results The in vivo results revealed that compared with the I/R group, the electroacupuncture pretreatment group showed significant decreased myocardial infarct size, as well as the increased indices of the cardiac function, including heart rate, mean arterial pressure, left ventricular systolic pressure and maximal rate for left ventricular pressure rising and declining (±dp/dt max). In addition, electroacupuncture pretreatment could inhibit the elevation of LDH and CK activities induced by I/R injury. The quantitative PCR (qPCR) results demonstrated electroacupuncture pretreatment could provide cardioprotection against myocardial I/R injury in rats with miR-214 up-regulation. In the meanwhile, in vitro, electroacupuncture pretreatment protected H9c2 cells from OGD-induced injury. Trans-fection of miR-214 mimic showed protective effects on OGD-induced injury to H9c2 cells by reducing apoptosis, decreasing LDH and CK activities, rescuing the OGD-induced Ca2+and down-regulating elevated protein levels of NCX1, BIM, CaMKIIδand CypD. Conclusions Our findings firstly demonstrated that electroacupuncture pretreatment promotes the expression of miR-214 in myocardial I/R injury and miR-214 contributes to the protective effect of electroacupuncture on myocardial I/R injury.

Experimental study on fasudil for increasing preserving effect on porcine DCD heart induced by ischemia / 重庆医学

Objective To evaluate the effects of fasudil on DCD hearts .Methods Sixteen domestic ,mixed‐breed male porcine [mean body weight (28 ± 3)kg] were randomly divide into experiment group and control group (n= 8 in each) .Experiment group :animals were sedated ,anesthetized and paralyzed .Mechanical ventilation with room air was provided .After thoroctomy ,cardiac arrest was established by bloodletting ,and then keeps at room temperature for additional 25 minutes (warm ischemia) .Hearts were perfused via the aortic root with fasudil (0 .1 mg/kg) enriched cold Stanford solution for coronary artery flushing ,rapidly excised and the aorta was cannulated .Hearts were subjected to isolated Langendorff perfusion (retrograde perfused with warm oxygenated autologous blood) afterwards ,and then underwent 20 minutes of equilibrium ,immersed in fasudil (0 .5 mg/kg) enriched cold Stan‐ford solution for 2 hours in situ cold preservation .Finally ,all the hearts were resuscitated with the warm oxygenated autologous blood perfusion .Control group :hearts from the animals experienced the same except for fasudil supplement .Left ventricular per‐formances were evaluated .Coronary blood flow ,myocardial infarction volume ,myocardial water content ,and myocardial enzyme were measured .Myocardial electron microscopic examinations were carried out as well .Results All the hearts from both groups were successfully resuscitated ,fasudil significantly decreased water content ,enzyme leakage (P< 0 .01 each versus control group) , and increased coronary blood flow (P< 0 .01 versus control group) .Left ventricular function were better preserved (P< 0 .01 each versus control group) .All hearts lacked severe necrosis as determined by tetrazolium staining ,myocardial infarction volume were decresed in experiment group (P< 0 .01 each versus control group) .Intracellular components retained various types of organelle in both group ,but still ultrastructural alterations in control group were more distinctive than in experiment group .Conclusion (1) Donor heart arrested by exsanguinations and plus 25 minutes warm ischemia ,could still be resuscitated with satisfactory result ;(2) The addition of fasudil to Stanford solution (before excise of graft ,during the cold preservation) ,might alleviate DCD heart I‐R in‐jury ,improving the effect of DCD heart preservation ,and hopefully ,might be a novel arsenal in clinical DCD heart transplantation in the future .

Cardioprotective effects of Guanxinshutong (GXST) against myocardial ischemia/reperfusion injury in rats / 老年心脏病学杂志（英文版）

Background The protective effects against reperfusion injury of cardioprotective drugs have recently been evaluated and found to be inadequate. Guanxinshutong (GXST), a combination of the traditional herb and Mongolian medicine, is effective and safe in treating angina pectoris in clinical trials. We assess the cardioprotective effects of GXST against myocardial ischemia and reperfusion (MI/R) injury in rats and explore its possible mechanism. Methods Forty-five male Sprague Dawley rats were randomized into three groups: non-MI/R group (Sham, n = 15), MI/R group treated with vehicle (Control, n = 15) and MI/R group treated with GXST (Drug, n = 15). MI/R was induced by ligation of the left anterior descending coronary artery (LAD) for 30 minutes, followed by 2/24 hour reperfusion in the Control and Drug groups. In the Sham group, the LAD was exposed without occlusion. GXST powder (in the Drug group) or saline (in the Control and Sham groups) were administered via direct gastric gavage from 7 day prior to surgery. Blood samples were collected from the carotid artery (10 rats each group) after 2 hours of reperfusion, to determine the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6) and intercellular adhesion molecule-1 (ICAM-1) using enzyme-linked immunosorbent assays. The animals were then sacrificed and the hearts were harvested for histopathology and western blot analysis. Infarct size was measured in the remaining five rats in each group after 24 hours reperfusion. Results GXST significantly decreased levels of TNF-α, IL-1β, IL-6, ICAM-1, apoptosis index (AI) and infarct size. GXST also obviously inhibited nuclear factor kappa B (NF-κB) activity when compared with the Control group (all P < 0.05). Conclusions GXST is effective in protecting the myocardium against MI/R injury in rats. Its possible cardioprotective mechanism involves inhibition of the inflammatory response and apoptosis following MI/R injury.

Application of MALDI-TOF MS in Renal Ischemia/Reperfusion(I/R) Injury Research / 医学研究杂志

Objective Proteomics changes from the proteserum which isolated from the rat model of renal ischemia/reperfusion(I/R) injury are detected and investigated by the matrix-assisted UV laser desorption ionization time of flight mass sperctra (MALDI-TOF MS). Methods After the establishment of rat renal ischemia-reperfusion model,the serum samples which we selected respectively in 6,12,24 hours after reperfusion in each group were detected by MALDI-TOF MS analysis. And the peptide fingerprint which existed differences in each group were analyzed to identify.SPSS13.0 software was used to the analysis the data. At the same time,we used Mascot Search to determine their nature in protein database. Results ①the serum which was analyzed by IMAC-Cu bead was detected and had statistically significant peptide fingerprint in the m/z 2481 Da.②the results obtained from peptide mass fingerprint (PMF) were analyzed by Mascot search program for protein identification. We identified it as rat fibrinogen fragment.Conclusion Fibrinogen in kidney ischemia/reperfusion (I/R) injury plays an important role.

Effects of Sodium 2,3-Dimercapto-1-propanesulfonate (DMPS) on ET-1 During Experimental Myocardial Ischemia-reperfusion Injury / 医学研究杂志

Objective To study the effects of DMPS on ET-1 during experimental myocardial ischemia-reperfusion (I/R) injury.Methods 20 New Zealand rabbits were randomly assigned to 2 groups: I/R group and DMPS protection group with 10 in each group. The blood sample was obtained through vien at different time (5 min before ischemia, the end of the ischemia period and 0.5h, 1h, 2h, 4h, 6h after reperfusion ) in each group.The serum concentrations of ET-1 were detected with radioimmunology method. Results The levels of ET-1 of serum and cardiac tissues increased after ischemia and reperfusion, and were significant different compared with that before ischemia(P0.05).Conclusions The changes of ET-1 were significant when myocardial I/R. DMPS may effectively effect the levels of ET-1 after myocardial ischemia and during I/R injury,and have protecfion of myocardium from ischemia and reperfusion injury.