异搏定对大鼠视网膜缺血再灌注损伤的保护作用

目的:在大鼠视网膜缺血再灌注损伤模型上,研究异搏定对视网膜缺血再灌注损伤后视网膜内细胞凋亡的影响。方法:60只大鼠随机分为对照组及异搏定处理组,每组30只。通过结扎大鼠左颈总动脉1h,然后再灌注。异搏定处理组腹腔注射异搏定1mg/kg,对照组腹腔注射生理盐水1mL/kg,分别检测1、6、12、24、48、72h各时段大鼠视网膜内的细胞凋亡,每时段大鼠各5只。结果:再灌注后,细胞凋亡主要出现于视网膜的神经节细胞层和内核层,大鼠视网膜在再灌注6h逐渐出现细胞凋亡,12h逐渐增加,24h细胞凋亡达到高峰,偶尔在外核层可见凋亡细胞。然后细胞凋亡逐渐减少,但是,到了术后72h,仍然可见视网膜内有细胞凋亡,而异搏定可以减少视网膜细胞凋亡的发生。结论:异搏定可以抑制再灌注后细胞凋亡的发生,对大鼠视网膜具有保护作用。

EFFECT OF ACUPUNCTURE ON NEURONAL APOPTOSIS AFTER FOCAL CEREBRAL ISCHEMIC REPERFUSION INJURY IN RATS

Objective To observe the impacts of acupuncture on cell-cycl ODK4） and neuronal death in hippocampal neurons in rats with focal cerebra e-related factors （cyclin D1, schemic reperfusion injury Methods Middle cerebral artery occlusion （MCAO） was used to establish the model of cerebral ischemic reperfusion injury. Western blot （WB） and flow cytometry （FCM） were applied to the tests of cell-cycle-related factors and apoptosis respectively. Results In 48 h of reperfusion, the expressions of cell-cycle-related factors （cyclin D1, CDK4） in hippocampal neurons and apoptosis were increased. In acupuncture group, the expressions of cyclin DI and CDK4 and apoptosis were reduced remarkably （P 〈 0.01 ）. Conclusion Acupuncture plays the protective role in cerebral ischemic reperfusion injury, which is contributed probably to the modulation of cell-cycle-related factors to inhibit apoptosis.

Anti-apoptotic effects of aspirin following cerebral ischemia-reperfusion injury in rats

BACKGROUND： The pharmacological effects of aspirin on apoptosis are complex. The underlying mechanisms have not been properly defined. OBJECTIVE： To observe the effect of different doses of aspirin on brain cell apoptosis following focal cerebral iscbemia-reperfusion injury （CIRI） in rats. DESING, TIME AND SETTING： A randomized, controlled, animal experiment, performed at the School of Medicine and Pharmaceutics, Jiangnan University between June and October 2006. MATERIALS： Twenty-six male, adult, Sprague Dawley rats （grade Ⅱ）, weighing 240-290 g, were obtained from Shanghai Experimental Animal Center, Chinese Academy of Sciences. Aspirin was provided by Sigma （USA）. METHODS： The rats were randomly divided into four groups： sham-operation （SO）, CIRI ＋ vehicle, CIRI ＋ aspirin （6 mg/kg）, and CIRI ＋ aspirin （60 mg/kg）. Rats in the lesion groups were intragastrically administrated saline, aspirin （6 mg/kg）, or aspirin （60 mg/kg）, respectively. MAIN OUTCOME MEASURES： The number of pyramidal neurons with normal appearance in the cerebral cortex at 24 mm from the midline; apoptotic cell death as measured by TUNEL; Bcl-2 and Bax protein localization was determined by immunohistochemistry; malondialdehyde （MDA） and super oxidation （SOD） content were determined by biochemistry method; adenosine triphosphate （ATP） content measured by capillary electrophoresis. RESULTS： Following CIRI, the following parameters were altered compared with sham-operated animals： the number of neurons with normal appearance was significantly reduced in the cerebral cortex; the number of apoptotic cells increased; Bax protein expression was enhanced; and the ratio between Bcl-2 and Bax decreased. In addition, MDA content increased significantly, whereas ATP content decreased （P 〈 0.01）. Aspirin ameliorated the loss of healthy pyramidal neurons. Both 6 and 60 mg/kg aspirin increased the ratio between Bcl-2 and Bax, with no significant difference between the treatment groups. In addition, 60 mg/kg aspirin decreased MDA content and increased ATP levels. However, 6 mg/kg aspirin did not have the same effect. CONCLUSION： Aspirin reduced the number of apoptotic cells following CIRI. These results suggest that the neuroprotective mechanism of aspirin could be related to elevated Bcl-2 protein levels or decreased Bax protein expression. The increase in the ratio of Bcl-2 to Bax appears to be a common anti-apoptotic mechanism of aspirin.

Effects of hypothalamic paraventricular nuclei on apoptosis and proliferation of gastric mucosal cells induced by ischemia/reperfusion in rats

AIM： To investigate the effects of electrical stimulation of hypothalamic paraventricular nuclei （PVN） on gastric mucosal cellular apoptosis and proliferation induced by gastric ischemia/reperfusion （I/R） injury. METHODS： For different experimental purposes, stimulating electrode plantation or electrolytic destruction of the PVN was applied, then the animals＇ GI/R injury model was established by clamping the celiac artery for 30 min and allowing reperfusing the artery for 30 rain, 1 h, 3 h or 6 h respectively. Then histological, immunohistochemistry methods were used to assess the gastric mucosal damage index, the gastric mucosal cellular apoptosis and proliferation at different times. RESULTS： The electrical stimulation of PVN significantly attenuated the GI/R injury at 30 min, i h and 3 h after reperfusion. The electrical stimulation of PVN decreased gastric mucosal apoptosis and increased gastric mucosal proliferation. The electrolytic destruction of the PVN could eliminate the protective effects of electrical stimulation of PVN on GI/R injury. These results indicated that the PVN participated in the regulation of GI/R injury as a specific area in the brain, exerting protective effects against the GI/R injury, and the protection was associated with the inhibition of cellular apoptosis and the promotion of gastric mucosal proliferation. CONCLUSION： Stimulating PVN significantly inhibits the gastric mucosal cellular apoptosis and promots gastric mucosal cellular proliferation. This may explain the protective mechanisms of electrical stimulation of PVN against GI/R injury.

PI 3-kinase pathway is responsible for antiapoptotic effects of atrial natriuretic peptide in rat liver transplantation

AIM： To investigate the in vivo effect of atrial natriuretic peptide （ANP） and its signaling pathway during orthotopic rat liver transplantation.METHODS： Rats were infused with NaCI, ANP （5 μg/ kg）, wortmannin （WH, 16μg/kg）, or a combination of both for 20 min. Livers were stored in UW solution （4℃） for 24 h, transplanted and reperfused. Apoptosis was examined by caspase-3 activity and TUNEL staining. Phosphorylation of Akt and Bad was visualized by Western blotting and phospho-Akt-localization by confocal microscopy.RESULTS： ANP-pretreatment decreased caspase-3 activity and TUNELopositive cells after cold ischemia, indicating antiapoptotic effects of ANP in vivo. The an- tiapoptotic signaling of ANP was most likely caused by phosphorylation of Akt and Bad, since pretreatment with PI 3-kinase inhibitor WM abrogated the ANP-induced reduction of caspase-3 activity. Interestingly, analysis of liver tissue by confocal microscopy showed translocation of phosphorylated Akt to the plasma membrane of hepa- tocytes evoked by ANP.CONCLUSION： ANP activates the PI-3-kinase pathway in the liver in vivo leading to phosphorylation of Bad an event triggering antiapoptotic signaling cascade in ischemic liver.

Edaravone inhibits apoptosis caused by ischemia/reperfusion injury in a porcine hepatectomy model

AIM： TO investigate the effect of E3-methyl-l-phe- nyl-2-pyrazolin-5-one （Edr） on hepatic ischemia-reper- fusion （I/R） injury and liver regeneration in a porcine hepatectomy model. METHODS： One hour ischemia was induced by occlud- ing the vessels and the bile duct of the right and median lobes. A 40% left hepatectomy was performed after re- perfusion. Six animals received Edr （3 mg/kg per hour） intravenously and six control animals received saline just before reperfusion. Remnant liver volume, hemody- namics, aspartate aminotransferase （AST）, alanine ami- notransferase, lactate dehydrogenase and lactic acid, were compared between the groups. The expression of transforming growth factor-β （TGF-β1） and toll-like receptor （TRL） mRNA in hepatic tissues was examined using reverse transcription polymerase chain reaction. Apoptosis was demonstrated by terminal deoxynucleo- tidyl transferase dUTP nick end labeling （TUNEL） stain- ing, respectively. RESULTS： Serum AS-I- （P = 0.029）, and toll like recep- tor 4 level （P = 0.043） were significantly lower after 3 hin animals receiving Edr. In addition, TUNEL staining in Edr-treated pigs showed significantly fewer hepatocytes undergoing apoptosis compared with control pigs. After 1 mo, all factors were non-significantly different between the two groups. CONCLUSION： Edr is considered to reduce hepatic injury in the early stage of I/R injury in a porcine model.

THE EFFECT OF ISCHEMIC RE-PERFUSION INJURY PLUS PARTICLE INFUSION EMBOLISM ON THE APOPTOSIS OF RATS WITH PANCREATIC CANCER

Objectives. In an attempt to develop new method of treating the end ormid stage pancreatic cancer, we examined the effect of ischemic re perfusion injury plus particle embolism on the pathology and cell apoptosis of pancreatic cancer in Sprague Dawely rats. Methods. 9 mg dimethylbeneanthracine (DMBA) were implanted directly into the parenchyma of pancreatic tail of Sprague Dawely rats. After establishment of tumor, the inferior splenic artery, a main supplying vessel to pancreatic tail was subjected to blockade and re opening for 30 min separately, then embolism particles were infused via the artery. Afterwards, artery was ligated. Pathological changes and cell apoptosis indicators (AI) of pancreatic cancer were observed by light microscopy and ISEL respectively 14 days after the operation. Results. The prevalence of pancreatic cancer among DMBA implanted rats evaluated 3 months to 4 months after implantation was 59%. The volumes of the tumor in positive control group (B), pancreatic ischemic group (C), pancreatic ischemic re perfusion injury group (D) were significantly larger than pancreatic ischemic re perfusion injury plus particle thrombus group (E) (P< 0.01). Thevolumes of the tumor in groups D, E were significantly smaller than that in group C (P< 0.01). There was a significant difference in tumor size between group Band group C (P< 0.01), but the difference was not significant between group D and group E (P >0.05). There was a significant infiltration of tumor tissue in group B rats, but strong inflammatory reaction was not noted. In groups C, D, E, alocalized tumor growth was observed; infiltration of inflammatory cells and proliferation of fibroblasts and connective fiber were obvious, and some of these fibers grew into cancer nests and separate the tumor. The above findings were most conspicuous in group E. There was a significant difference in AI between group E (13.7±1.5)and other groups (P< 0.01), with the difference being also significant between group C(4.3±2.4), D (8.5±1.1)and group B (1.2±0.8)(P< 0.01), and between group C and group D (P< 0.01) or between group D and group E (P< 0.01). In the samples of group A, the apoptotic cells were not found. Conclusions. Pancreatic ischemic re perfusion injury plus particle thrombus can cause significant infiltration of inflammatory cells in tumor tissues thereby limiting its growth, and inducing cell apoptosis of pancreatic cancer. This effect is superior to either pancreatic ischemia alone or pancreatic ischemia plus re perfusion injury.

Moderate hypothermia prevents neural cell apoptosis following spinal cord ischemia in rabbits

Paraplegia is a disastrous complication after operations of descending and thoracoabdominal aortic aneurysm. Re- gional hypothermia protects against spinal cord ischemia although the protective mechanism is not well know. The objective of this study is to examine whether hypothermia protects the spinal cord by preventing apoptosis of nerve cell and also investigate a possible mechanism involved in hypothermia neuroprotection. Cell apoptosis with necrosis was evident in the spinal cord 24 h after 30 min of ischemia. Moderate hypothermia decreased the incidence of apoptotic nerve cells. Both cell apoptosis and necrosis were attenuated by hypothermia. p53 expression increased and bcl-2 expression declined after ischemia, while hypothermia mitigated these changes. This study suggests that apoptosis contributes to cell death after spinal cord ischemia, and that moderate hypothermia can prevent nerve cell apoptosis by a mechanism associated with bcl-2 and p53 genes.

MicroRNA-15a/b are up-regulated in response to myocardial ischemia/reperfusion injury

Objective Several studies have indicated that miR-15a,miR-15b and miR-16 may be the important regulators of apoptosis.Since attenuate apoptosis could protect myocardium and reduce infarction size,the present study was aimed to find out whether these miRNAs participate in regulating myocardial ischemia reperfusion （I/R） injury.Methods Apoptosis in mice hearts subjected to I/R was detected by TUNEL assay in vivo,while flow cytometry analysis followed by Annexin V/PI double stain in vitro was used to detect apoptosis in cultured cardiomyocytes which were subjected to hypoxia/reoxygenation （H/R）.Taqman real-time quantitative PCR was used to confirm whether miR-15a/15b/16 were involved in the regulation of cardiac I/R and H/R.Results Compared to those of the controls,I/R or H/R induced apoptosis of cardiomyocytes was significantly iucreased both in vivo （24.4％ ± 9.4％ vs.2.2％ ± 1.9％,P ＜ 0.01,n =5） and in vitro （14.12％ ±0.92％ vs.2.22％ ± 0.08％）.The expression of miR-15a and miR-15b,but not miR-16,was increased in the mice I/R model,and the results were consistent in the H/R model.Conclusions Our data indicate miR-15 and miR-15b are up-regulated in response to cardiac I/R injury,therefore,down-regulation of miR- 15a/b may be a promising strategy to reduce myocardial apoptosis induced by cardiac I/R injury.

Effects of acupuncture plus mild hypothermia on apoptosis-related factors in rats with cerebral ischemia-reperfusion

Objective： To investigate the effect of acupuncture plus mild hypothermia on neurological function impairment score, cerebral infarct size and apoptosis-related factors in cerebral ischemia reperfusion injury（CIRI） rats. Methods： Sixty healthy male Sprague-Dawley（SD） rats were routinely reared for 1 week. Ten rats were randomly selected as the sham operation group and 10 rats as the blank control group, while the remaining 40 rats were subjected to preparing the middle cerebral artery occlusion（MCAO） model by modified filament occlusion method. The 40 MCAO rats were further randomly divided into a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, with 10 rats in each group. Rats in the sham operation group, the blank control group and the model group did not accept treatment except binding; rats in the acupuncture group received acupuncture treatment; rats in the mild hypothermia group received mild hypothermia treatment; rats in the acupuncture plus mild hypothermia group received acupuncture and mild hypothermia treatment. 72 h after the treatment, neurological function impairment score was performed; the infarct area ratio was determined by 2,3,5-tripheyl tetrazolium chloride（TTC） staining; apoptosis of brain cells was observed by TUNEL method; the expressions of Bcl-2, Bax and Caspase-3 were detected by immunohistochemistry.Results： Compared with the blank control group and the sham operation group, the neurological function impairment score, cerebral infarct area ratio, apoptosis, and the expressions of Bax and Caspase-3 in the model group were significantly increased, while the expression of Bcl-2 was significantly decreased, and there were significant between-group differences（all P〈0.05）. After the treatment, there were statistically significant differences among the treatment groups in the neurological function impairment score, cerebral infarct area ratio and apoptosis in the ischemic side of rats, as well as the expressions of Bcl-2, Bax and Caspase-3（all P〈0.05）, and from the figures, tables and statistical analysis, it was found that a better tendency in the acupuncture plus mild hypothermia group than the acupuncture group or mild hypothermia group. Conclusion： Acupuncture plus mild hypothermia can protect the brain cells by improving neurological function impairment, decreasing cerebral infarct area ratio, reducing the number of apoptotic cells in the ischemic area and regulating the expressions of apoptosis related proteins to inhibit apoptosis.

Effect of acupuncture plus mild hypothermia on MAPK/ERK pathway of brain tissues in rats with cerebral ischemia-reperfusion injury

Objective： To observe the protective effect of acupuncture plus mild hypothermia on brain tissues in rats with cerebral ischemia-reperfusion injury （CIRI）, and the influence on protein expression levels of phosphorylated Raf-1, MEK-2 and ERK3/2 in the mitogen-activated protein kinase （MAPK）/extracellular regulated protein kinases （ERK） pathway, and to explore the mechanism of acupuncture plus mild hypothermia therapy for the ischemic stroke. Methods： Ninety Sprague-Dawley （SD） rats were randomly divided into a blank control group, a sham operation group, a model group, an acupuncture group, a mild hypothermia group and an acupuncture plus mild hypothermia group, 15 rats in each group. Except the rats in the blank control group, the remaining rats were used to prepare the middle cerebral artery occlusion （MCAO） models according to the modified occlusion method using lines, while only the occlusion lines were inserted without blocking the brain arteries of rats in the sham operation group. When the vital signs of rats were stable, rats in the blank control group did not receive any intervention; rats in the sham operation group and the model group received fastening without treatment; rats in the acupuncture group, the mild hypothermia group, and the acupuncture plus mild hypothermia group were treated with the corresponding therapeutic methods. 72 h later, observed neurologic injury score, evaluated infarction area ratio by 2,3,5-tripheyl tetrazolium chloride （TTC） staining, determined apoptosis by TUNEL assay, and measured the phosphorylated Raf-1, MEK-2 and ERK3/2 protein expression levels in rat ischemic hippocampal tissues by Western blot assay. Results： Compared with the blank control group and the sham operation group, after modeling, the neurologic injury score, infarction area ratio and apoptotic cells were increased, and phosphorylated Raf-1, MEK-2 and ERK：1/2 protein expression levels were significantly increased in the model group; the differences were statistically significant （P〈0.05 or P〈0.01）. Compared with the model group, after acupuncture or mild hypothermia therapy, neurologic injury score and infarction area ratio were decreased; apoptotic cells and phosphowlated Raf-1, MEK-2 and ERK1/2 protein expression levels were significantly decreased; the differences were statistically significant （P〈0.05 or P〈0.01）. Compared with the acupuncture group, neurologic injury score and phosphorylated Raf-1, MEK-2 and ERK3/2 protein expression levels were decreased in the acupuncture plus mild hypothermia group; differences between the groups were statistically significant （P〈0.05 or P〈0.01）. Compared with the mild hypothermia group, phosphorylated Raf-1, MEK-2 and ERK1/2 protein expression levels decreased in the acupuncture plus mild hypothermia group, and differences were statistically significant （P〈0.01）. Conclusion： Acupuncture or mild hypothermia therapy can improve neurologic injury, reduce infarction area and apoptosis, which brought about protective effect on the brain tissues, in the MCAO model. The protective effect of acupuncture plus mild hypothermia group is the strongest. The mechanism may involve the MAPK/ERK pathway, by reducing the phosphorylated Raf-l, MEK-2 and ERK：1/2 protein expression levels.

Effects of Acupuncture Pretreatment on Ischemic Cardiac Muscle Cell Apoptosis and Gene Expression in Ischemia-reperfusion Rats

Objective： To investigate the protective effects of acuptmcture pretreatment on ischemic myocardium, the protective mechanism of acupuncture pretreatment on ischemic myocardium was explored by observing the cardiac muscle cell apoptosis and the expression of HSP70 mRNA of ischemia-reperfusion injury rats treated with acupuncture pretreatment. Methods： Sixty-four Wistar rats were randomly divided into eight groups： control group, sham surgery group, ischemia-reperfusion group, ischemia pretreatment group, manual acupuncture pretreatment group （once a day）, electroacupuncture pretreatment group （once a day）, manual acupuncture pretreatment group （twice a day）, and electroacupuncture pretreatment group （twice a day）. The reperfusion model of rat myocardial ischemia was made. Expression of HSP70 mRNA was assayed by in situ hybridization, and cell apoptosis by TUNEL. Results： Compared with those in the control group and the sham surgery group, the apoptosis and the expression of HSP70 mRNA were increased in the ischemia-reperfusion group. Compared with those in the ischemia-reperfusion group, the cardiac muscle cell apoptosis was decreased and the HSP70 mRNA was increased in the rats treated with acupuncture pretreatment; meanwhile, acupuncture pretreatment twice a day had stronger effects than acupuncture pretreatment once a day and ischemia pretreatrnent. Conclusion： Acupuncture pretreatment can inhibit the cardiac muscle cell apoptosis, and up-regulate the expression of HSP70 mRNA in ischemia-reperfusion rats. Acupuncture pretreatment twice a day has stronger effects than pretreatment once a day.

Effects of Chinese Herbal Medicine Serum on the Apoptosis of Sinoatrial Node Cells Induced by Simulated Ischemia-reperfusion

Objective:To study the effect of Chinese herbal medicine Kangxin Fumai Granule(康心复脉颗粒 Granule for heart diseases) serum on the primary cultured sinoatrial node(SAN) cell apoptosis induced by simulated ischemia-reperfusion(IR).Methods:The SAN cells removed from SAN tissue of neonatal Wistar rats were cultured and purified with differential attachment and 5'-bromodeoxyuridine(BrdU) treatment.Simulated IR model was adopted.The obtained cells were morphologically observed with inverted microscopy.By using the method of serum pharmacology,the cell apoptosis was measured with TUNEL staining qualitatively and with flow cytometry quantitatively.Results:Three kinds of cells were observed in the cultured SAN cells:spindle,triangle and irregular.The spindle cells comprised the greatest proportion.The SAN cells in the model group showed moderate positive brown staining in the nucleus,and the apoptosis rate increased significantly compared to that in the control group(P<0.01).While the SAN cells in the Kangxin Fumai Granule high-dose group did not demonstrated positive staining in the nucleus,and the apoptosis rate decreased significantly compared to that in the model group(P<0.05).Conclusion:Of the cells cultured from SAN,the spindle cells were pacemaker cells of SAN in rats.Blockade and/or inhibition of the SAN cell apoptosis might be one of the important mechanisms of Kangxin Fumai Granule in preventing and treating sinoatrial injury induced by simulated IR.

Antioxidant properties of magnesium lithospermate B contribute to the cardioprotection against myocardial ischemia/reperfusion injury in vivo and in vitro

OBJECTIVE： To determine the cardioprotective ef- fect of magnesium lithospermate B （MLB） on myo- cardial ischemia/reperfusion （MI/R） injury and to in- vestigate the antioxidant potential in vivo and in vitro. METHODS： MI/R injury was induced by the occlu- sion of left anterior descending coronary artery for 30 min followed by reperfusion for 3 h in rats. After reperfusion, hearts were harvested to assess infarct size, histopathological damages, the levels of superoxide dismutase （SOD）, catalase （CAT）, glutathione peroxidase （GPx）, reduced glutathione （GSH） and malondialdehyde （MDA）. Blood samples were collected to determine serum levels of creatine kinase-MB （CK-MB）, cardiac troponin （cTnl） and lactate dehydrogenase （LDH）. Furthermore, simulatedischemia/reperfusion （SI/R） injury in vitro was established by oxygen and glucose deprivation （OGD） for 2 h followed by 24-hour recovery period in cardiomyocytes. The activity of LDH in the cultured su- pernatant and the levels of intracellular reactive oxygen species （ROS）, SOD and MDA in cardiomyo- cytes were also measured. Finally, cardiomyocytes apoptosis was determined with flow cytometry. RESULTS： MLB significantly limited infarct size, ameliorated histopathological damages and prevented leakage of CK-MB, cTnl and LDH. Additional- ly, SOD, CAT, GPx and GSH activities were notably increased by MLB, along with the MDA content decreased as compared with the model group in rats. In vitro study, MLB also decreased LDH activity in the cultured supernatant, increased SOD activity in cardiomyocytes, reduced intracellular ROS and MDA levels, and significantly suppressed cardiomyocytes apoptosis. CONCLUSION： MLB possessed remarkably cardioprotective effects on MI/R injury in vivo and in vitro. The protection of MLB may contribute to its antioxidant properties.

Salusins protect myocardium against ischemic injury by alleviating endoplasmic reticulum stress

Salusins are regulatory peptides that affect cardiovascular function. We previously reported that salusin-a and -β protected cultured cardiomyocytes from serum deprivation-induced cell death through upregulating glucose-regulated protein 78 （GRP78）, an endoplasmic reticulum （ER） resident protein whose overexpression acts as a marker and suppressor of ER stress. The present study examined whether salusin-α and -β inhibit ER stress in ischemic myocardium. In a rat model of myocardial infarction created by ligating the left anterior descending coronary artery （LAD）, salusin-α or -β was intravenously injected at 5 or 15 nmol kg-1 15 min prior to 2 h of LAD occlusion. The high dose of salusin-α and -β3 significantly improved heart function and hemodynamics in LAD-occluded rats, but had no effects in sham-operated rats. The arrhythmias caused by LAD oc- clusion were markedly attenuated by salusin-α and -β. The apoptotic rate in ischemic myocardium was reduced from 31.5%±3.7% to 19.8%±2.2% and 12.3%±2.2%, and the infarct size was reduced from 53.4%±4.0% of the risk area to 26.5%±9.7% and 23.7%±8.9% by 15 nmol kg-1 salusin-α and -β, respectively. Furthermore, salusin-α and -β prevented the ac- tivation of GRP78 and ER stress-specific apoptotic effectors caspase-12 and CHOP （C/EBP homologous protein）, and attenu- ated the reduction of an ER stress-associated antiapoptotic protein Bcl-2 in ischemic cardiac tissue. The salusins also inhibited the ER stress induced by tunicamycin in cultured rat H9c2 cardiomyocytes. These results indicate that salusins protect myo- cardium against ischemic injury by inhibiting ER stress and ER stress-associated apoptosis.

Combinatorial effects of conception and governor vessel electroacu-puncture and human umbilical cord blood-derived mesenchymal stem cells on pathomorphologic lesion and cellular apoptosis in rats with cerebral ischemia/reperfusion

OBJECTIVE: To investigate the combinatorial effects of conception and governor vessel electroacupuncture(EA) and human umbilical cord blood-derived mesenchymal stem cells(HUCB-MSCs) on pathomorphologic lesion and cellular apoptosis in rats with cerebral ischemia/reperfusion. METHODS: With the HUCB-MSCs isolated, cultured and identified and the models of cerebral ischemia-reperfusion established, the HUCB-MSCs of passage three were intracranially transplanted and the EA at conception and governor vessels was applied. The pathomorphologic lesion by hematoxylin-eosin staining and the cellular apoptosis by terminal deoxynucleotidyl transferase-mediated nick-end labeling method around the ischemic focus were observed. RESULTS: The cultured adherent HUCB-MSCs exhibited a spindle shape and expressed MSC-specific markers, with the cell purity and proliferation rate significantly increasing after the primary passage. HE staining showed that there were no pathological changes observed in the sham surgery group. However, in the PBS transplantation group, degeneration and necrosis of a great number of nerve cells were seen. In both the HUCB-MSCs transplantation group and the HUCB-MSCs transplantation + EA group, reparative changes of the pathomorphism of the tissue were found. Both combination treatment and simple MSCs treatment were able to improve the pathomorphorlogic lesion following cerebral ischemia and reduce the abnormal TUNEL-positive numbers, with former better than latter. CONCLUSION: HUCB-MSCs improve pathological lesions and inhibit the cellular apoptosis around the cerebral ischemic area. EA at conception and governor vessels also improve pathological lesion and inhibit the cellular apoptosis in rats treated with HUCB-MSCs transplantation, which effects were superior to that of simple HUCB-MSCs transplantation.

Effects of Electroacupuncture on Hippocampal and Cortical Apoptosis in A Mouse Model of Cerebral Ischemia-reperfusion Injury

Objective: To observe the effects of electroacupuncture on hippocampal and cortical apoptosis in a mouse model of cerebral ischemia-reperfusion injury. Methods: Mouse models established by repeated cerebral ischemia-reperfusion, followed by electroacupuncture at Shenshu, Geshu, and Baihui points. The control group mice were intragastrically administered Hydergine. On day 1 and 7 post-treatment, hippocampal and cortical apoptosis was detected by terminal-deoxynucleotidyl transferase mediated dUTP nick-end labeling (TUNEL), and apoptosis images in the hippocampal CA1 zone and cortical area were analyzed. Results: In the model group, apoptotic cells were detected one day after treatment and some cellular fibers were disarrayed. By day 7 post-treatment, there was an increase in the number of apoptotic cells in the hippocampal CA1 region. In addition, there were apoptotic cells in the cortical area, the cortical layers were thinner with localized neuronal loss and sieve-like lymphocyte infiltration, as well as glial cell proliferation and visible infarct lesions. However, in the Hydergine and electroacupuncture groups, there was a small number of apoptotic cells. At 7 days post-treatment in the model group, field number, numerical density on area, and surface density were increased. However, in the Hydergine and electroacupuncture groups these parameters were decreased (P<0.01), with a significant difference between the two treatment groups (P<0.01). Conclusion: Electroacupuncture treatment inhibited apoptosis and provided neuroprotection.