Effect of Ginsenoside—Rb1 on Cardiomyocyte Apoptosis after Ischemia and Reperfusion in Rats

Summary The effect of ginsenoside Rb₁ on cardiomyocyte apotosis after ischemia (30 min) and reperfusion (6 h) in rats was observed. The ischemia/reperfusion heart model was established by ligating left anterior descending branch of coronary artery in Wistar rats. The apoptotic cardiomyocytes were examined under transmission electron microscopy and counted byin situ nick end labeling (TUNEL) method and light microscopy. Results showed that (1) The apoptotic cardiomyocytes were found in ischemic regions in the ischemia/reperfusion group, but not in the sham-operating group under transmission electron microscopy; (2) The number of apoptotic cells were 134.45±45.61/field in the ischemia/reperfusion group, 0/field in the sham-operating group and 51.65±13.71/field in the ginsenoside Rb₁-treated group. The differences were significant among the three groups (P<0.01). It was concluded that myocardial ischemia-reperfusion could induce cardiomyocyte apoptosis, and ginsenoside Rb₁ could significantly inhibit cardiomyocyte apoptosis induced by ischemia-reperfusion in rats, indicating that ginsenoside Rb₁ could inhibit cardiomyocyte apoptosis induced by ischemia-reperfusion, thus alleviating ischemia-reperfusion injury. GUAN Li, female, born in 1967, Doctor in Charge This project was supported by a grant from the Natural Science Foundation of Hubei Province (Serial No. 2000J050).     

Effect of Shenfu on the Expression of Bcl-2 and Bax of Intestinal Mucosal Epithelial Cells after Ischemic Reperfusion in Rats

Objective To study the effect of Shenfu on the expression of bax and bcl-2 in small intestinal mucosal epithelial cells after ischemic reperfusion injury and explore the effect of Shenfu on small intestinal mucosal epithelial cells apoptosis. Methods 36 SD rats were randomized into three groups, each consisting of 12 rats： Sham group （S group）, Ischemic reperfusion group （IR group）, Shenfu group （SF group）, Ischemic reperfusion models were made by ligated the superior menseneric artery for 1 hour followed by 2 hrs reperfusion. Histological mucosal damage in each group was graded according to Chiu＇s score. Immunohistochemistry detected the expression of bax and bcl-2, and obtained the optical density （OD） value using a color image pattern analysis system, and then calculated the ratio of bcl-2/bax. TUNEL method measured apoptotic intestinal mucosal epithelial cells, and calculated the apoptotic index. Results There were edema and epithelial impairment in part of villus and the integral was higher in SF group than S group （P〈0.05）, but lower than IR group （P〈0.01）. Compared IR group with S group and SF group, the average OD value of bcl-2 and bax had significant statistic difference （P〈0.01）, and the average OD value of bcl-2 in SF group was higher than in S group （P〈0.05）, meanwhile bcl-2/bax was significantly lower in IR group than in S group and SF group （P〈0.01）, but that was higher in SF group than in S group （P〈0.05）. Apoptotic index in IR group was significantly higher than that in other two groups （P〈0.01）, and that in SF group was higher than in S group （P〈0.05）. Conclusion Shenfu can enhance the expression of bcl-2, decrease the expression of bax, meanwhile increase the ratio of bcl-2/bax, inhibit small intestinal mucosal epithetlial cells apoptosis, and protect small intestine mucosal epithetlial after ischemic reperfusion.     

海马微量注射β淀粉样蛋白对Bax／Bcl—2的表达的影响

探讨Ｂａｘ／Ｂｃｌ－２基因表达在β－淀粉样蛋白脑内致凋亡机制中的作用。方法用微量注射器将Ａβ１－４０注射到大鼠右侧海马ＣＡ１区诱发Ａβ在脑内该区域的沉积，７ｄ后，用免疫组化ＳＡＢＣ法检测Ｂａｘ／Ｂｃｌ－２的表达。结论Ａβ脑内沉积诱导神经元凋亡的机制可能与其增强了Ｂａｘ的表达有关。     

Bcl—2、Bax基因与肺癌

Bcl- 2 ( B- cell lymphoma/leukemia- 2 gene)和Bax ( Bcl- 2 associated x protein)基因是近年来发现的细胞凋亡相关基因 ,它们与肿瘤的发生、发展及预后有密切的关系。有关这方面的研究已成为肿瘤领域的热门课题。本文就 Bcl- 2、Bax基因与肺癌的发生、发展、病理学特征及预后的关系作一综述。1　Bcl- 2基因的结构与功能　　 Bcl- 2基因是 Tsujimoto等 (1) 于 1 984年在人类滤泡性淋巴细胞瘤 1 4号和 1 8号染色体易位断裂点t ( 1 4∶ 1 8) ( q32 ;q2 1 )上发现的。其正常定位点为1 8q2 1。Bcl- 2基因有 3个外显子 ,全长 2 30 kb,c D…     

视网膜母细胞瘤Bcl—2和Bax基因蛋白质表达

目的：研究凋亡及凋亡调控基因Ｂｃｌ－２／Ｂａｘ和视网膜母细胞瘤（Ｒｅｔｉｎｏｂｌａｓｔｏｍａ，ＲＢ）的发生、发展及退化的关系。方法：收集ＲＢ标本，对其分别进行Ｂｃｌ－２和Ｂａｘ基因的蛋白质免疫组织化学染色。对其表达情况和染色强度进行观察。结果：①Ｂｃｌ－２在分化型ＲＢ中表达比较好；②Ｂａｘ在未分化型和分化型中表达都比较好。结论：①分化型和未分化型ＲＢ中都有Ｂｃｌ－２／Ｂａｘ基因蛋白表达；②随ＲＢ恶     

Effects of 2-APB on Store-operated Ca^2＋ Channel Currents of Hepatocytes after Hepatic Ischemia／Reperfusion Injury in Rats

Summary The effects of hepatic ischemia/reperfusion (I/R) injuries on hepatocellular viability and store-operated calcium current (Isoc) in isolated rat hepatocytes and the effects of 2-APB on store-operated calcium current (Isoc) in isolated rat hepatocytes after hepatic ischemia/reperfusion injuries were studied. Hepatic ischemia and reperfusion injury model was establised and whole cell patch-clamp techniques were used to investigate the effects of 2-APB on Isoc. The results showed that ischemia/reperfusion injuries could significantly reduce hepatocellular viability and further increase Isoc in hepatocyte and 2-APB (20, 40, 60, 80, 100 μmol/L) produced a concentration-dependent decrease of Isoc with IC₅₀ value of 64.63±10.56 μmol/L (n=8). It was concluded that ischemia/ reperfusion injuries could reduce hepatocellular viability, probably through increased Isoc in hepatocytes and 2-APB had a protective effect on ischemia/reperfusion-induced liver injury, probably though inhibiting Isoc. Huang Changzhou, male, born in 1965, M. D., Ph.D. This project was supported by the National Natural Science Foundation of China (No. 30270532), the Trans-Century Excellent Talent Development Plan Fund of Ministry of Education of China (Official Letter No. 2002-48) and Shuguang Program Project of Shanghai Educational Committee (No. 02SG20).     

Nimodipine Modulates Bcl-2 and Bax mRNA Expression after Cerebral Ischemia

In order to explore whether the member of Bcl-2 gene family, for example, Bcl-2 and Bax, are induced after cerebral ischemia, and whether expression of genes can be modulated by calcium-antagonist, the rat cerebral ischemic models were made by occluding left middle cerebral artery. The expression of Bcl-2 and Bax mRNA was measured by RT-PCR method. After middle cerebral artery occlusion (MCAO), the expression of both Bcl-2 and Bax mRNA were induced. Level of Bcl-2 mRNA increased steadily and level of Bax mRNA increased gradually at first, reached a peak after 24 h, then decreased slowly. After administration of nimodipine, Bcl-2 mRNA was up-regulated in the hippocampus 6 and 24h after ischemia, while Bax mRNA was down-regulated 6 and 24 h after ischemia. Focal cerebral ischemia can induce proto-oncogenes to express, which was associated with apoptosis. Calcium-antagonist can up-regulate Bcl-2 mRNA and down-regulate Bax mRNA.The increased ratio of Bcl-2 and Bax mRNA may contribute to the antiapoptic effect of nimodipine.The study indicates that pharmacological modulation of Bcl-2 family member expression could become a new stratehgy to manage neuronal damage.     

Bcl—2、Bax及M30在寻常疣中的表达

目的：研究细胞凋亡及其相关基因在寻常疣皮损中的表达及意义。方法：采用免疫组化SP法检测了29例建党疣皮损中Bcl-2,Bax及M30的表达。结果：M20均阴性，9例（31％）表皮角质形成细胞Bax阳性表达，仅6例（21％）基底细胞Bcl-2染色弱阳性。结论：寻常疣的增殖性改变可能不是由于表皮角质形成细胞凋亡降低引起的。     

Effect of Ginsenoside-Rb_1 on Cardiomyocyte Apoptosis after Ischemia and Reper fusion in Rats

It had been demonstrated by clinical trials andanimal experiments that Ginseng and its extractginsenoside could alleviate cardiomyocyte ischemia,restrict myocardial infarction size,and exertmarked protection effect on myocardial reperfusioninjury[1- 3] .More recentresearch revealed thatcar-diomyocyte apoptosisplays a key role in myocardialischemia- reperfusion injury[4 ] ,and ginsenoside ex-erted an inhibitory effect upon cardiomyocyteapoptosis in ischemia- reperfusion injury[5] .Ex-tracted gi…     

The Effect of Nitric Oxide／Endothelins System on the Hepatic Ischemia／Reperfusion Injury

In a hepatic ischemia/reperfusion( I/R) mod-el,we used three tool drugs,L- arginine( L- Arg) ,L- nitro- arginine methyl ester( L- NAME) and en-dothelins( ET) receptor antagonist TAK- 0 4 4 ( C4 5H51N9Na2 O11S) ,to regulate nitric oxide ( NO) /ETbalance and observe itsinfluence on hepatic I/R in-jury.1　 MATERIALSAND METHODS1 .1　Animal Grouping and Model EstablishmentSeventy- eight Wistar rats ( weighing 2 90 g-35 0 g,both sexes) were divided into operationcontrol group ( S…     

黄体酮对大鼠缺血再灌注脑组织细胞凋亡及Bcl-2、Bax表达的影响

目的:探讨黄体酮对缺血再灌注脑损伤的保护机制.方法:采用SD大鼠局灶性脑缺血再灌注模型(MCAO),将48只SD大鼠随机等分为6组:假手术组、脑缺血再灌注组、溶剂(DMSO)对照组、黄体酮(PROG)预防组、PROG治疗组、PROG防治组,应用免疫组织化学和原位末端标记(ISEL)法检测脑组织细胞凋亡及凋亡相关蛋白Bcl-2、Bax的表达情况.结果:假手术组每高倍视野下凋亡细胞数为1.88±0.25,缺血再灌注组为41.38±3.85, DMSO组为38.13±5.69,预防组为22.88±2.70,治疗组为25.63±2.93, 防治组为20.88±2.30;缺血再灌注组每高倍视野下Bcl-2蛋白阳性细胞数为9.50±1.69, DMSO组为10.25±2.61,预防组为 17.13±1.13,治疗组为16.63±1.30,防治组为23.50±1.93;缺血再灌注组每高倍视野下Bax蛋白阳性细胞数为28.13±2.75, DMSO组为27.75±3.06,预防组为 14.25±1.83,治疗组为13.00±1.85,防治组为11.38±1.41.各PROG处理组(预防组、治疗组及防治组3项指标与缺血再灌注组、DMSO组之间比较,差异均有统计学意义(P＜0.05.结论:PROG可抑制脑神经细胞中Bax表达,上调Bcl-2表达,从而抑制脑细胞凋亡,发挥脑保护作用.     

Study on the effects of losartan on cardiomyocyte apoptosis and gene expression after ischemia and reperfusion in vivo in rats

Along with the thrombolysis therapy, PTCAand the coronary artery bypass graft are used inthe treatment of acute myocardial infarction(AMI), these method played the important rolesfor restoring the myocardial perfusion, rescuingagonal cardiomyocyte or reducing the extension ofthe myocardial infarctlon and protecting the cardiacfunction, but ischemia/reperfusion can cause themyocardial injury. Therefore the study of the myocardial ischemia and reperfusion injury become theclinical hotspot. It …     

缺血后处理对缺血再灌注心肌细胞凋亡及Bcl-2和Bax蛋白表达的影响

目的:研究缺血后处理对缺血再灌注心肌细胞凋亡及Bcl-2和Bax蛋白表达的影响.方法:24只大耳白兔随机分成4组:假手术组(S)、缺血再灌注组(IR)、缺血预适应组(IP)和缺血后处理组(IPO).S组(6只):开胸只穿线不结扎血管,麻醉维持120 min;IR组(6只):结扎30 min,再灌注120 min;IP组(6只):结扎5 min,再灌注5 min,反复3次诱导缺血预适应,之后再按IR组操作;IPO组(6只):结扎30 min后, 再灌注10 s、缺血10 s, 反复6次,再灌注120 min.应用TUNEL法观察各组心肌细胞凋亡,应用免疫组化SP法观察Bcl-2和Bax蛋白的表达.结果:与IR组相比,IP组和IPO组心肌梗死范围减小,心肌细胞的凋亡指数降低(P＜0.05),Bcl-2/Bax比例升高(P＜0.05),而IP组和IPO组间差异无统计学意义(P＞0.05).结论:缺血后处理可以减轻再灌注心肌细胞的凋亡,这一过程可能是通过升高Bcl-2/Bax比值来实现的.     

Melatonin通过影响Bcl-2／Bax比率抑制再灌注后大鼠肝细胞凋亡

目的 探讨再灌注损伤对大鼠肝细胞凋亡的影响。Mel对大鼠肝再灌注细胞凋亡的影响作用及其机制。方法 150只健康雄性Wistar大鼠（质量190-210g，6-7周龄），随机分为褪黑素处理组（Mel）、酒精溶媒对照组（Alc）和生理盐水对照组（NS）。建立肝缺血再灌注损伤模型，缺血均为60min。之后每组分别按再灌注后0．5、1、6、12及24h采集标本。M组（20mg／kg）于缺血前30min腹腔注射melatonin；A组采取与Mel组相同浓度的酒精液，N组则注射同比例的生理盐水。测定血清天门冬酸氨基转移酶（AST）进行测定；对肝组织进行Bcl-2及Bax免疫组化染色；并在此基础上进行透射电镜观察肝细胞核及细胞器。结果 Mel组在再灌注后的6及12h时点AST均显著低于Alc及NS对照组（aP〈0．05），且Alc组与NS组相比差异无显著性。Mel组在再灌注后各时点的Bcl-2染色的阳性细胞率显著高于Alc及NS对照组（aP〈0．05），且各时点内Alc组与NS组相比差异无显著性。Mel组在再灌注后1，6，12及24h时点的Bax染色的阳性细胞率显著低于Alc及NS对照组（cP〈0．05）。且各时点内Alc组与NS组相比差异无显著性。Mel组再灌注后6，12及24h的Bcl-2／Bax值显著高于Alc组和NS组（aP〈0．05）。且上述每时点内的Alcohol组与N．s．组相比差异无显著性。电镜观察结果在Alcohol组及N．s对照组发现了普遍的肝细胞凋亡现象，相同时点Mel组则未发现肝细胞凋亡的发生。结论 外源性Mel可以抑制再灌注后血清天门冬酸氨基转移酶（AST）。增强肝细胞Bcl-2蛋白的表达，抑制Bd-2蛋白的表达，通过提升Bcl-2／Bax值来抑制再灌注后肝细胞凋亡的发生。     

细胞凋亡和Bcl-2、Bax基因在兔肺缺血再灌注损伤中的作用

目的:探讨细胞凋亡在兔肺缺血再灌注损伤中的作用以及Bcl-2和Bax基因的调控.方法:健康日本大耳白兔48只,随机分为对照组与肺缺血再灌注损伤1 h、3 h和5 h组.复制在体肺缺血再灌注损伤模型.采用TUNEL法观测肺组织细胞凋亡;予免疫组化及原位杂交技术检测肺组织细胞Bcl-2、Bax蛋白和基因表达.结果:随着再灌注时间的延长,凋亡指数(AI)及Bax蛋白、BaxmRNA水平进行性升高;Bcl-2蛋白及Bcl-2mRNA先升高而后下降(均P＜0.01).凋亡指数(AI)与Bax蛋白及Bax mRNA之间均呈显著正相关(r分别=0.926,0.933;均P＜0.01),而与Bcl-2蛋白/Bax蛋白及Bcl-2mRNA/BaxmRNA的比值呈显著负相关(r分别=-0.836,-0.879;均P＜0.01).结论:肺组织细胞凋亡参与了肺缺血再灌注损伤的发生,Bcl-2/Bax的比值下降是促进凋亡的重要因素.     

针刺对发育期脑损伤大鼠脑组织细胞凋亡及Bcl-2、Bax蛋白表达的影响

目的观察针刺疗法对宫内缺血脑损伤大鼠脑组织细胞凋亡及其凋亡相关蛋白Bcl-2、Bax表达的影响,为针刺疗法应用于发育期脑损伤提供实验依据。方法采用结扎孕鼠双侧子宫动脉,完全阻断血供25 min娩出胎鼠,制成宫内窘迫脑损伤模型,将成功的模型随机分为针刺组和模型组,每组各8只;正常组剖宫产取出胎鼠,8只;其余胎鼠丢弃。正常组和模型组不治疗,针刺组于出生后7~30 d行针刺治疗,每日针刺1次,不留针。30 d实验结束后断头取脑,运用TUNEL法检测脑细胞凋亡,SABC法测脑组织Bcl-2、Bax蛋白表达。结果正常组大鼠脑细胞凋亡数极少,而模型组较高,针刺组介于两者之间,3组差异有统计学意义(P<0.05);Bcl-2染色指数正常组或针刺组比模型增加,差异有统计学意义(P<0.05);Bax染色指数正常组或针刺组比模型组降低,差异有统计学意义(P<0.05)。结论针刺可以减少宫内窘迫脑损伤大鼠脑组织神经细胞凋亡,提高Bcl-2蛋白的表达,降低Bax蛋白的表达,这可能是应用针刺疗法治疗发育期脑损伤的机制之一。     

大鼠局灶性脑缺血再灌注后神经细胞凋亡和Bcl-2蛋白表达的改变及银杏叶提取物的保护作用

目的研究大鼠局灶性脑缺血再灌注后神经细胞凋亡和Bcl-2蛋白表达的变化及银杏叶提取物对其表达的影响.方法制造大鼠大脑中动脉缺血再灌注模型.40只Wistar雄性大鼠被随机分为A假手术组、B缺血组、C小剂量治疗组和D大剂量治疗组.于术前30min及术后1 h分别给予银杏叶提取物20mg/kg和40mg/kg腹腔内注射.应用TTC染色及HE染色观察梗死体积及缺血坏死程度,应用免疫组化染色及POD法检测Bcl-2蛋白表达及凋亡细胞数.结果C、D两组梗死体积明显小于B组(P＜0.01),D组梗死体积小于C组(P＜0.05);C、D两组凋亡细胞数明显少于B组(P＜0.01),D组凋亡细胞数少于C组(P＜0.05);C、D两组Bcl-2蛋白表达明显高于B组(P＜0.01),D组Bcl-2蛋白表达高于C组(P＜0.05).结论银杏叶提取物可通过上调Bcl-2蛋白表达,减少神经细胞凋亡,对脑缺血再灌注损伤起保护作用,疗效与剂量有关.     

Effects of Ethyl Pyruvate on Myocardial Apoptosis and Expression of Bcl-2 and Bax Proteins after Ischemia-reperfusion in Rats

In order to study the effects of ethyl pyruvate on cardiomyocyte apoptosis following ischemia/reperfusion （I/R） in vitro and the expression of Bcl-2 and Bax proteins, isolated rat hearts were perfused in a Langendorff model. Twenty-four rats were randomly divided into 3 groups （n=8 in each group）： control group was perfused for 120 min. In the I/R group, after 30 min stabilization the injury was induced by 30 min global ischemia followed by 60 min reperfusion. Ethyl pyruvate （EP） group was set up with the same protocol as I/R group except that it was supplied with 2 mmol/L EP 15 rain before ischemia and throughout reperfusion. Myocardial malonaldehyde （MDA） content was measured. Myocardial apoptotic index （AI） was tested by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling （TUNEL） method. The expression of anti-apoptotic protein Bcl-2 and pro-apoptotic protein Bax in cardiac myocytes was detected by immunohistochemistry. As compared with control group, the content of MDA, myocardial AI and the expression of Bcl-2, Bax proteins were increased significantly in I/R group, but the content of MDA, myocardial AI and the expression of Bax protein were decreased obviously and the expression of Bcl-2 protein was up-regulated in EP group （P〈0.05）. These results demonstrate that EP could inhibit apoptosis of cardiac myocytes possibly via alleviating oxidative stress, up-regulating Bcl-2 and down-regulating Bax proteins.     

不同类型颈动脉粥样斑块Bcl-2/Bax的基因表达

目的观察Bcl-2/Bax凋亡调控基因在不同类型的颈动脉粥样斑块中的表达,探讨该基因表达与粥样硬化斑块稳定性和脑缺血症状的关系.方法选取人体颈动脉内膜剥脱术标本42例,根据病理学分为稳定型(19例)和不稳定型(23例)两组,8例正常腹主动脉及其分支作为对照组.Bcl-2/Bax抗凋亡基因表达分别采用免疫组化和原位杂交进行检测.结果免疫组化和原位杂交检测Bcl-2在不稳定型斑块中分别表达20和9例,在稳定型斑块中表达11和4例(P＜0.05);在平滑肌细胞、内皮细胞和泡沫细胞均有表达.Bax在不稳定性斑块中分别表达18和11例,在稳定性斑块中分别表达8和5例(P＜0.05);在泡沫细胞、平滑肌细胞均有表达.Bcl-2/Bax在细胞成分聚集的肩区,表达强度在不稳定型斑块中明显高于稳定型斑块(P＜0.01).结论Bcl-2/Bax在不稳定型斑块的表达均高于稳定型斑块,其在颈动脉粥样硬化不同时期的表达可能是调控颈动脉粥样硬化发病机制之一.