生长激素对大鼠阿霉素心肌病心肌细胞凋亡及P53的影响

目的探讨生长激素(growth hormone,GH)对阿霉素(adriamycin,ADR)心肌病大鼠心肌细胞凋亡的影响及对凋亡相关基因P53蛋白表达的影响.方法30只雄性Wistar大鼠,体重200～250 g,随机分为3组,对照组、ADR组和GH联用ADR组.用原位末端标记法标记凋亡的心肌细胞,用免疫组织化学方法检测P53蛋白的表达.结果与对照组比较,ADR组心肌细胞凋亡指数升高的差异有统计学意义(P＜0.05),GH联用ADR组细胞凋亡指数低于ADR组(P＜0.05),ADR组P53蛋白表达明显高于对照组(P＜0.05)GH联用ADR组P53蛋白表达明显低于ADR组(P＜0.05).结论阿霉素可使大鼠心肌细胞发生凋亡,同时使P53蛋白表达升高.GH干预治疗可减少阿霉素心肌病的心肌细胞凋亡.     

卡维地洛对外源性羟自由基诱导心肌细胞凋亡的影响

目的 :观察卡维地洛 (Carvedilol)对外源性羟自由基诱导心肌细胞凋亡的影响。方法 :采用培养的第 2代心肌细胞 ,随机分成 4组 :1.正常对照组 :仅用DMEM培养 ;2 .羟自由基组 :FeSO410 0 μmol·L- 1 +H2 O2 10 0 μmol·L- 1 ;3.卡维地洛组 :卡维地洛终浓度为 1μmol·L- 1 ;4 .卡维地洛 +羟自由基组 :卡维地洛 1μmol·L- 1 +FeSO4 10 0 μmol·L- 1 +H2 O2 10 0 μmol·L- 1 。观察心肌细胞存活率和形态学 ,流式细胞仪检测细胞凋亡率。结果 :1.羟自由基组心肌细胞存活率降低 ,和对照组比较有显著性差异(P <0 0 5 ) ;卡维地洛 +羟自由基组细胞存活率较羟自由基组高 (P <0 0 5 )。 2 .羟自由基组凋亡率较高 ,明显高于对照组 (P <0 0 5 ) ;卡维地洛 +羟自由基组的细胞凋亡率显著低于羟自由基组 (P <0 0 5 )。3.羟自由基组凋亡心肌细胞呈特征性超微结构及AnnexinV+ PI- 荧光染色。结论 :卡维地洛能减轻外源性羟自由基诱导的心肌细胞凋亡。     

不同剂量卡维地洛干预对心肌梗死后心力衰竭大鼠心肌细胞凋亡和凋亡相关基因表达的影响

目的 :探讨不同剂量卡维地洛 (CAR)对心肌梗死 (MI)后大鼠心肌细胞凋亡的影响及CAR治疗充血性心力衰竭 (CHF)的作用机制。方法 :将雄性Waistar大鼠前降支结扎 ,于术后 1周开始分别予大剂量CAR[HCAR组 ,6 0mg/ (kg·d) ]和小剂量CAR[LCAR组 ,6mg/ (kg·d) ]干预 7周 ,观察不同剂量CAR对大鼠血流动力学参数、心肌细胞凋亡、Fas、Fas配体 (FasL)和Bcl 2mRNA表达的影响。结果 :CAR可改善心功能指标 ,降低心肌细胞凋亡指数、Fas及FasL的mRNA水平 ,上调Bcl 2表达 ,其中对心肌细胞凋亡指数、Fas及Bcl 2mRNA表达的影响以HCAR组明显。结论 :CAR可有效地减少MI后心肌细胞凋亡 ,防治CHF ,效果以大剂量明显 ,这可能与其对凋亡相关基因表达影响的差异有关     

红景天苷对H2O2诱导乳鼠心肌细胞凋亡的抑制作用

目的 探讨红景天苷对H2O2诱导乳鼠心肌细胞凋亡的影响及其可能的作用机制.方法 实验于2006年11月-2007年7月在辽宁医学院药理实验室完成.在原代培养的SD大鼠乳鼠心肌细胞上建立H2O2损伤模型,观察不同剂量红景天苷(10 μg/mL、30 μg/mL、90 μg/mL)对心肌细胞凋亡率、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、乳酸脱氢酶(LDH)活性、细胞内钙离子[Ca2 ]i浓度的影响.将原代培养的心肌细胞分为5组:空白组、模型组及红景天苷(Sal)不同剂量组(10 μg/mL、30 μg/mL、90 μg/mL),每组6孔.观察检测细胞培养液中MDA含量、SOD活性、LDH含量;利用Fura-2/AM负载心肌细胞,荧光分光光度计检测细胞内[Ca2 ]i;利用Annexin VFITC/PI双标记染色后做流式细胞双色分析检测心肌细胞凋亡率.结果 与模型组比较不同剂量红景天苷组均可以降低培养液中MDA含量增加,SOD活性和降低LDH活性,减少[Ca2 ]i,减少心肌细胞凋亡率(P<0.01).结论 红景天苷对心肌细胞凋亡有抑制作用,其作用机制可能与其抗脂质过氧化、抑制细胞内钙超载有关.     

芹菜素对大鼠阿霉素性心肌损伤的影响

目的观察芹菜素(Apigenin,API)对大鼠阿霉素(Adriamycin,ADR)性心肌损伤的影响。方法将雄性SD大鼠40只随机分为5组:对照组、阿霉素组、阿霉素+芹菜素(低剂量)、阿霉素+芹菜素(中剂量)组、阿霉素+芹菜素(高剂量)组。检测光镜下的心肌细胞形态学、心肌酶CK-MB以及心肌细胞TLR4的表达。结果与对照组相比较,阿霉素组大鼠光镜下的心肌损伤明显,CK-MB以及TLR4的表达升高(P<0.01);与阿霉素组相比较,阿霉素+芹菜素各剂量组的心肌损伤减轻,CK-MB以及TLR4的表达下降,尤以中剂量组、高剂量组显著(P<0.01)。结论芹菜素对大鼠阿霉素性心肌损伤具有保护作用,可能与其对心肌细胞TLR4的表达的影响有关。     

Kr(u)ppel样因子4对阿霉素诱导大鼠心肌细胞H9C2凋亡的影响

目的 探讨阿霉素对大鼠心肌细胞H9C2 凋亡的影响及Kr(u)ppel样因子4过表达对阿霉素所致H9C2细胞凋亡的影响.方法 采用台盼蓝染色检测细胞存活率,流式细胞术检测细胞凋亡率,采用Western blotting检测Kr(u)ppel样因子4及凋亡相关基因多聚二磷酸腺苷核糖聚合酶的表达.结果 阿霉素能够诱导H9C2细胞凋亡,采用5 μmol/L阿霉素处理时,24 h变化最为明显;Kr(u)ppel样因子4过表达之后,细胞凋亡率较转空栽体组更高,达87.90%.阿霉素能够诱导Kr(u)ppel样因子4在H9C2细胞中表达增加;Kr(u)ppel样因子4过表达组多聚二磷酸腺苷核糖聚合酶裂解片段的表达水平明显上调.结论 Kr(u)ppel样因子4能够促进阿霉素诱导的大鼠心肌细胞H9C2凋亡,其作用机制与多聚二磷酸腺苷核糖聚合酶有关.     

重组人肝再生增强因子抑制实验性肝纤维化明胶酶A基因表达

目的　了解重组人肝再生增强因子 (hALR)对实验性肝纤维化明胶酶A(MMP2 )基因表达的影响。方法　建立CCl4中毒性及人血白蛋白免疫损伤性两种大鼠肝纤维化模型 ,在造模的同时给予不同剂量 (10 ,5 0 μg·kg-1·d-1)的hALR。在不同的时间点留取大鼠肝组织标本 ,提取总RNA ,用逆转录定量聚合酶链反应 (RT PCR)测定MMP2 的基因表达水平。结果　在两种模型中两个剂量hALR预防组大鼠肝组织MMP2 的基因表达水平在模型形成的不同阶段均明显低于模型组 ;高剂量hALR组大鼠肝组织MMP2 的基因表达水平均明显低于低剂量组。结论　重组人肝再生增强因子可能有抑制大鼠实验性肝纤维化MMP2 基因表达的作用     

色满卡林对H/R损伤心肌细胞凋亡及bax、bcl-2表达的影响

目的 探讨色满卡林(CRK)对培养大鼠心肌细胞缺氧复氧(H/R)损伤后心肌细胞凋亡及凋亡相关基因bax、bcl-2蛋白表达的影响.方法 原代培养SD大鼠心肌细胞随机分为5组:正常对照组:不施加任何处理因素正常培养;H/R损伤组:缺氧2 h、复氧30 min;CRK各浓度组:分别加入终浓度为2.5、5、10 μmol/L的CRK后均即刻缺氧2 h、复氧30 min.各组细胞经AV-PI双标记后应用流式细胞仪检测心肌细胞凋亡率;Western blot法检测bax、bcl-2蛋白表达.结果 H/R组心肌细胞凋亡率与正常对照组比较明显增高,bax蛋白表达水平升高、bcl-2蛋白表达下降;CRK各浓度组与H/R组相比,心肌细胞凋亡率明显降低、bax蛋白表达水平下调、bcl-2蛋白表达水平上调,且有一定剂量依赖性.结论 CRK可抑制H/R致培养大鼠心肌细胞的凋亡,其作用可能是通过下调bax、上调bcl-2蛋白表达实现的.     

猕猴桃果王素对大鼠阿霉素心肌损伤的保护作用及其机制

目的:观察猕猴桃果王素对大鼠阿霉素心肌损伤的保护作用,并探讨其作用机制。方法:腹腔注射阿霉素建立大鼠阿霉素心肌损伤模型。观察大鼠体重变化,计算各组大鼠肝肺的干重/湿重、心脏重量/体重及死亡率;测定各组大鼠脑钠肽、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)、铜-锌-超氧化物歧化酶(Cu-Zn-SOD)值;TUNEL法检测细胞凋亡;免疫组化法检测Cu-Zn-SOD、bax、bcl-2蛋白水平表达,RT-PCR检测Cu-Zn-SOD、bax、bcl-2mRNA水平表达。结果:猕猴桃果王素可使bcl-2、Cu-Zn-SOD在蛋白、mRNA水平表达增加,bax在蛋白、mRNA水平表达减少;增强GSH-Px、CAT、Cu-Zn-SOD活力,改善大鼠心功能,减少死亡率。与对照组比较,阿霉素加果王素高剂量组差异有统计学意义(P<0·05),其疗效呈剂量依赖性。结论:猕猴桃果王素对大鼠阿霉素心肌损伤有明显疗效,其机制可能与抗氧化损伤和抑制细胞凋亡等作用有关。     

心肺血管病杂志2003年(第22卷)索引

文题索引(文题关键词以汉语拼音为序 )A阿霉素　生长激素对阿霉素中毒大鼠心肌细胞的抗凋亡作用　 (吴旭辉 )　 ( 4 ) :2 36B白细胞　白细胞和单核细胞计数与急性心肌梗死临床预后的关系　 (贾士杰 )　 ( 4 ) :2 2 4白细胞介素 6　切割球囊血管成型术对C 反应蛋白白细胞介素 6及细胞间粘附分子的影响　 (马晓静 )　 ( 4 ) :2 0 4白细胞介素 18　内毒素血症大鼠心肌TLR4与IL 18mRNA表达的变化　 (张德明 )　 ( 2 ) :10 9白血病　慢性粒细胞白血病合并多浆膜腔积液 1例　 (高云阁 )　 ( 2 ) :70病人教育　支气管哮喘患者个体化教育的效…     

Granulocyte Colony-Stimulating Factor Reduces Cardiomyocyte Apoptosis and Improves Cardiac Function in Adriamycin-Induced Cardiomyopathy in Rats

Background: Cardiomyocyte apoptosis reportedly participates in the occurrence and progression of dilated cardiomyopathy (DCM). Recent studies have shown that granulocyte colony-stimulating factor (G-CSF) enhances bone marrow cells migration to the damaged heart in the DCM model and improves the ultrastructure of the cardiomyocyte in adriamycin (ADR) induced DCM. However, its influence on cardiac pump function and cardiomyocyte apoptosis has not been studied. Methods and materials: Wistar Rats were randomly grouped into control, ADR, ADR+PBS, ADR+G-CSF group (n = 10). ADR (2.5 mg/kg, 6 times for 2 weeks) was administered intraperitoneally in all rats except the control group. After 2 weeks, the rats in ADR+G-CSF group were injected with G-CSF (50 microg/kg/day for 8 days) subcutaneously. Cardiac function was evaluated by echocardiogram and cardiac catheterization after 4 weeks. Cardiomyocytes apoptosis and apoptosis-related protein Fas were detected by in situ terminal deoxynucleotidyl transferase assay (TUNEL method) and Western blot, respectively. Results: The ADR and ADR+PBS groups showed significant deteriorations of left ventricular functions and high cardiomyocyte apoptosis index, as well as high Fas expressions. Meanwhile, the ADR+G-CSF group showed significant improvement in LV function, inhibition of cardiomyocyte apoptosis compared with the ADR and ADR+Phosphate-Buffered Saline PBS group. The Fas protein expression was remarkably attenuated as well. Conclusion: Our results suggest that administration of G-CSF inhibited cardiomyocyte apoptosis and Fas protein expression and contributes to improving cardiac pump function in vivo in ADR induced DCM rat model.     

Staurosporine诱导小鼠心肌细胞凋亡模型的构建

目的 :Staurosporine可诱导不同细胞类型的细胞凋亡 ,但尚缺乏其对小鼠心肌细胞作用影响的报道。本研究旨在观察 Staurosporine是否也可引起小鼠心肌细胞的凋亡 ,并对其模型进行分子水平的鉴定。方法 :用 Stau-rosporine处理原代培养的小鼠心肌细胞的直接刺激后 ,观察心肌细胞的形态学变化、DNA片段、半胱天冬蛋白酶(caspase) -3活性、细胞存活率以及细胞膜完整性。结果 :1Staurosporine处理过的心肌细胞具有明显的凋亡形态学特征 :细胞的皱缩、核的浓缩及 DNA梯改变。2 Staurosporine4μmol/ L 对心肌细胞作用 8h后 ,与正常对照组比较细胞存活率降至 3 1.2 % (与对照组比 ,P<0 .0 1) ,且细胞存活率与 Staurosporine的作用浓度和作用时间呈依赖关系 ;同时测定细胞培养液中能反映细胞膜完整性的胞浆内容物乳酸脱氢酶 (L DH)水平 ,发现在 Staurosporine作用 1～ 8h的各个时间点 ,L DH的漏出水平最高未超过 10 % (与对照组相比 ,P>0 .0 5) ;这种高的细胞死亡率与低的细胞膜的损伤率 ,恰恰反映了细胞死亡的性质是凋亡性死亡。3 Staurosporine能激活心肌细胞内的 Caspese-3的活性 ,当使用广谱的 Caspese抑制剂 ZVAD-fmk预处理培养的小鼠心肌细胞后 ,能够有效的阻止上述细胞凋亡的发生 ,从而避免了 Staurospor     

Apoptosis in hypertensive heart disease: a clinical approach

PURPOSE OF REVIEW: It is widely accepted that there are two principal forms of cell death, namely, necrosis and apoptosis. According to the classical view, necrosis is the major mechanism of cardiomyocyte death in cardiac diseases. RECENT DEVELOPMENTS: In the past few years observations have been made showing that cardiomyocyte apoptosis occurs in diverse conditions including hypertensive heart disease, and that apoptosis may be a contributing cause of loss and functional abnormalities of cardiomyocytes in this condition. SUMMARY: This review will summarize recent evidence demonstrating the potential contribution of cardiomyocyte apoptosis to heart failure in hypertensive patients. In addition, some strategies aimed to detect and prevent apoptosis of cardiomyocytes will be considered.     

高糖高脂对培养成年大鼠心肌细胞损伤观察及机制初探

目的建立成年大鼠心肌细胞培养模型,观察给予高糖、高脂刺激后是否发生心肌细胞损伤和凋亡,并分析其可能机制。方法将分离所得成年大鼠心肌细胞接种入培养皿后随机分为2组进行培养。正常对照组：以M199培养基（Media199）加入5mmol／L葡萄糖与20mmol／L甘露醇作为正常对照培养基培养细胞;高糖高脂组：以M199加入高糖（25mmol／L葡萄糖）高脂（600μmol／L软脂酸）作为培养基培养模拟糖尿病。培养48h后,分别收集培养基上清与细胞,进行心肌损伤和凋亡指标测定。结果高糖高脂组乳酸脱氢酶（LDH）,细胞凋亡蛋白caspase-3、caspase-8、caspase-9活性显著升高,与正常对照组比较,差异均具有统计学意义（P均〈0．01）。结论在培养的成年大鼠心肌细胞,高糖、高脂可引起心肌细胞损伤和凋亡,这种凋亡与easpase-8和caspase-9依赖的凋亡途径有关。     

吡格列酮保护缺氧复氧诱导乳鼠心肌细胞凋亡的离子通道机制

目的观察吡格列酮对缺氧复氧大鼠心肌细胞凋亡的保护作用,并探讨保护作用与ATP敏感性钾通道开放的关系。方法原代培养的SD乳鼠心肌细胞,分为8组:溶媒组、缺氧复氧组、0.1μmol/L吡格列酮组、1 μmol/L吡格列酮组、2μmol/L吡格列酮组、2μmol/L吡格列酮+30μmol/L HMR-1098组、2μmol/L吡格列酮+0.5mmol/L 5-羟基葵酸盐(5-HD)组、2μmol/L吡格列酮+30μmol/L HMR-1098+0.5 mmol/L 5-HD组。利用膜联蛋白-V与碘化丙啶双染法及Hoechst 33258染色法检测心肌细胞凋亡。结果缺氧复氧组乳鼠心肌细胞发生明显凋亡,经吡格列酮预处理后减少心肌细胞凋亡(P<0.05);与0.1μmol/L、1μmol/L及2μmol/L吡格列酮组比较,5-HD削弱吡格列酮抑制心肌细胞凋亡的作用(P<0.05),HMR-1098无作用(P>0.05)。结论吡格列酮对缺氧复氧乳鼠心肌细胞凋亡具有保护作用,线粒体ATP敏感性钾通道的开放可能介导了吡格列酮的保护作用。     

缬沙坦对阿霉素致心肌损伤大鼠心肌结构改变的影响

目的研究缬沙坦对阿霉素(adriamycin,ADR)致大鼠心功能损伤及心肌结构改变的保护作用。方法大鼠腹腔注射ADR(20mg/kg)造成心脏损伤模型。观察缬沙坦对ADR致心肌损伤大鼠体重、左心室功能、血清超氧化物歧化酶(SOD)、肌酸激酶同工酶(CK-MB)和丙二醛(MDA)的变化。结果①ADR组大鼠血清CK-MB活性和MDA含量及左心室舒张末期压(LVEDP)较对照组明显增加;血清SOD、左心室收缩压(LVSP)、左心室等容收缩期压力变化速率最大值( dp/dtmax)显著降低;电镜下可见,ADR组大鼠心肌细胞严重损伤,心肌纤维溶解,肌质网扩张空泡化,线粒体水肿,嵴断裂。②缬沙坦 阿霉素组大鼠CK-MB活性和MDA含量及LVEDP显著降低;SOD和 dp/dtmax显著升高;电镜下可见,局灶性的心肌细胞肌浆溶解;散在性的心肌细胞空泡样变。结论缬沙坦可减轻自由基对心脏的损伤,改善ADR致心肌损伤的毒性反应,并能有效地防治心功能损伤。     

MicroRNA-214在缺氧所致心肌细胞损伤过程中的作用

目的:探讨MicroRNA-214(miR-214)表达的变化与低氧诱导心肌细胞损伤的关系。方法:实时荧光定量PCR技术检测miR-214在心肌细胞低氧损伤中表达的变化。根据培养环境和有无使用miR-214的特异性抑制剂,将心肌细胞分为常氧培养组、常氧培养+miR-214抑制组、低氧培养组及低氧培养+miR-214抑制组。MTT比色法测定各组细胞活力,流式细胞仪(FCM)检测各组细胞的凋亡情况,酶法测定各组细胞的乳酸脱氢酶(LDH)漏出情况。结果:低氧培养后,心肌细胞内的miR-214表达相对常氧培养的对照组有所增高(P0.05),并呈时间依赖关系,低氧培养24 h,miR-214表达相对常氧培养的对照组可增高(1.9±0.1)倍。低氧培养会降低心肌细胞的存活率、增加心肌细胞凋亡及LDH漏出,miR-214抑制后,缺氧造成的上述损伤有所减轻。结论:miR-214的表达水平升高与低氧诱导的心肌细胞损伤有关,可能作为对抗缺氧损伤的靶点和诊断指标。     

Developmental expression of myostatin in cardiomyocytes and its effect on foetal and neonatal rat cardiomyocyte proliferation

OBJECTIVES: Myostatin, a member of the transforming growth factor-beta (TGF-beta) family, plays a key role in skeletal muscle myogenesis by limiting hyperplastic and hypertrophic muscle growth. In cardiac muscle, myostatin has been shown to limit agonist-induced cardiac hypertrophic growth. However, its role in cardiac hyperplastic growth remains undetermined. The aim of this study was to characterise the expression of myostatin in developing myocardium, determine its effect on cardiomyocyte proliferation, and explore the signalling mechanisms affected by myostatin in dividing cardiomyocytes. METHODS: We used quantitative PCR and Western blotting to study the expression of myostatin in cardiomyocytes isolated from rat myocardium at different developmental ages. We determined the effect of recombinant myostatin on proliferation and cell viability in dividing cardiomyocytes in culture. We analysed myostatin's effect on cardiomyocyte cell cycle progression by flow cytometry and used Western blotting to explore the signalling mechanisms involved. RESULTS: Myostatin is expressed differentially in cardiomyocytes during cardiac development such that increasing expression correlated with a low cardiomyocyte proliferation index. Proliferating foetal cardiomyocytes, from embryos at 18 days of gestation, expressed low levels of myostatin mRNA and protein, whereas isolated cardiomyocytes from postnatal day 10 hearts, wherein the majority of cardiomyocytes have lost their ability to proliferate, displayed a 6-fold increase in myostatin expression. Our in vitro studies demonstrated that myostatin inhibited proliferation of dividing foetal and neonatal cardiomyocytes. Flow cytometric analysis showed that this inhibition occurs mainly via a block in the G1-S phase transition of the cardiomyocyte cell cycle. Western blot analysis showed that part of the mechanism underpinning the inhibition of cardiomyocyte proliferation by myostatin involves phosphorylation of SMAD2 and altered expressions of the cell cycle proteins p21 and CDK2. CONCLUSIONS: We conclude that myostatin is an inhibitor of cardiomyocyte proliferation with the potential to limit cardiomyocyte hyperplastic growth by altering cardiac cell cycle progression.     

Cardiomyocyte apoptosis in hypertensive cardiomyopathy

It is widely accepted that there are two principal forms of cell death; namely, necrosis and apoptosis. According to the classical view, necrosis is the major mechanism of cardiomyocyte death in cardiac diseases. However, in the past few years observations have been made showing that cardiomyocyte apoptosis occurs in diverse conditions and that apoptosis may be a contributing cause of the loss and functional abnormalities of cardiomyocytes with important pathophysiological consequences. In this regard, although a number of formal proofs are pending, it is conceivable that cardiomyocyte apoptosis may be an important variable in the clinical evolution of hypertensive cardiomyopathy. This review summarizes recent evidence demonstrating that cardiomyocyte apoptosis is abnormally stimulated in the heart of animals and humans with arterial hypertension. In addition, the potential mechanisms of cardiomyocyte apoptosis in hypertension and its detrimental impact on cardiac function will be addressed. Finally, the perspectives of strategies aimed to detect and modulate apoptosis of cardiomyocytes in hypertensive cardiomyopathy will be considered.