依达拉奉保护H9c2心肌细胞对抗阿霉素的心肌毒性作用

目的探讨新型的自由清除剂依达拉奉(edaravone,EDA)能否保护H9c2心肌细胞对抗阿霉素(doxorubicin,DOX)引起的损伤。方法应用DOX(5μmol·L-1)处理H9c2心肌细胞建立DOX心肌毒性损伤模型。CCK-8比色法测定细胞存活率;Hoechst 33258核染色法观察细胞凋亡的形态学和数量改变;双氯荧光素(DCFH-DA)染色荧光显微镜照像检测细胞活性氧(ROS)水平;罗丹明123(Rh123)染色荧光显微镜照像测定线粒体膜电位(MMP);Western blot法测定caspase-3蛋白的表达水平。结果应用20、40、80μmol·L-1EDA分别预处理H9c2心肌细胞60 min,可明显地抑制5μmol·L-1DOX引起的细胞毒性,使细胞存活率升高,其中40μmol·L-1EDA的保护作用最大;应用40μmol·L-1EDA分别预处理心肌细胞30、60、90、120 min,可明显地抑制DOX引起的细胞毒性,其中预处理60 min的保护作用最大;此外,在5μmol·L-1DOX处理H9c2心肌24 h前,应用40μmol·L-1EDA预处理60 min可明显抑制DOX引起的心肌损伤作用,表现为抑制DOX引起的细胞内ROS生成增多及抑制DOX的致细胞凋亡作用(使凋亡细胞数目减少和cleaved caspase-3表达下调)和MMP的损伤作用。结论EDA能保护H9c2心肌细胞对抗DOX诱导的心肌毒性,此保护作用可能与其抑制ROS生成及减轻DOX对MMP的损伤有关。     

硫化氢对多柔比星致心肌细胞凋亡的保护作用

目的:研究硫化氢对多柔比星引起的心肌细胞凋亡的保护作用及其机制.方法:以硫氢化钠(NaHS)作为供体,先用不同浓度NaHS处理心肌细胞,再加入多柔比星(DOX)致其凋亡.应用胎盘蓝染色测量细胞的存活率;应用Annexin V和PI双染流式术检测其凋亡情况;应用比色法检测凋亡相关蛋白CASPASE-3和CASPASE-9的活性.结果:分别以DOX(5μmol·L-1)+ NaHS(50μmol·L-1)、DOX(5μmol·L-1)+ NaHS(100 μmol·L-1)、DOX(5μmol·L-1)+ NaHS( 200 μmol·L-1)浓度处理的各组细胞存活率均较单用DOX组(5μmol·L-1)增高;以DOX(5μmol ·L -1)+ NaHS(200 μmol·L-1)处理的细胞凋亡率较单用DOX组(5μmol·L-1)降低,以DOX(5μmol·L-1)+ NaHS(200μmol·L-1)处理的细胞的caspase-3和caspase-9活性较单用DOX组(5μmoloL-1)降低,各项指标差异均有统计学意义(P＜0.05).结论:硫化氢可以保护DOX引起的心肌细胞损伤,抑制DOX引起的心肌细胞凋亡.     

化合物LH-17对谷氨酸导致原代皮层神经元凋亡的保护作用

研究化合物LH-17对谷氨酸诱导原代皮层神经元凋亡的保护作用及其机制。L-谷氨酸钠(0.1,0.5和1.0 mmol·L-1)处理神经元30 min进行造模,化合物处理组在加入L-谷氨酸钠前给予美金刚(1μmol·L-1)或不同剂量的LH-17(0.1,1和10μmol·L-1)预孵育1 h再造模;换正常神经元培养基继续培养24 h后,MTT法检测细胞存活率,流式细胞仪检查细胞凋亡和坏死率,琼脂糖电泳分析凋亡后DNA lad-der情况;Fura-2对细胞内钙进行染色,荧光显微镜观察LH-17瞬时给药后细胞内钙离子浓度的影响,试剂盒分析NO水平。结果表明,L-谷氨酸钠浓度依赖性地诱导神经元发生凋亡和坏死,LH-17能抑制L-谷氨酸钠对细胞的毒性作用,减少凋亡小体,降低DNA断裂程度;L-谷氨酸能引起细胞内钙浓度大幅增加,用LH-17预处理后,增加幅度明显降低(P<0.05);同时,LH-17逆转了L-谷氨酸导致的细胞内NO浓度增高。以上结果提示,LH-17可能通过抑制神经元内钙超载,及下游NO浓度增高而起到减少谷氨酸引起的兴奋性氨基酸毒性的作用。     

柚皮苷保护H9c2心肌细胞对抗阿霉素诱导的心肌毒性

目的探讨柚皮苷(NRG)能否保护H9c2心肌细胞对抗阿霉素(DOX)诱导的心肌毒性。方法应用DOX处理H9c2心肌细胞建立DOX心肌损伤模型。CCK-8比色法测定细胞存活率;谷胱甘肽试剂盒检测GSSG/(GSSG+GSH)的比值;Hoechst 33258核染色法观察细胞凋亡的形态学和数量改变;Western blot法测定葡萄糖调节蛋白78(GRP78)的表达水平;双氯荧光素(DCFH-DA)染色荧光显微镜摄片检测细胞活性氧(ROS)水平;罗丹明123(Rh123)染色荧光显微镜照像测定线粒体膜电位(MMP)。结果 DOX在3⁷μmol·L-1浓度范围内处理H9c2心肌细胞24 h,呈剂量依赖性降低细胞存活率,其中5μmol·L-1DOX能明显引起心肌细胞损伤,表现为使细胞存活率下降近50%,并呈时间依赖性上调GRP78蛋白的表达;1μmol·L-1NRG预处理60min可明显抑制5μmol·L-1DOX引起的心肌毒性作用,表现为细胞存活率升高,GSSG/(GSSG+GSH)的比值下降,凋亡细胞数目减少,GRP78表达被抑制,细胞内ROS产生及MMP丢失减少。结论 NRG能保护心肌细胞对抗DOX诱导的心肌细胞损伤,保护作用可能与其抗氧化应激及内质网应激有关。     

黄连对小鼠成纤维细胞L929的毒性实验研究(英文)

黄连是一种广泛应用的传统中药,历来被认为无毒,很少人注意到黄连的毒性及其在细胞和基因水平上相关的毒性机制。本实验以体外培养的小鼠成纤维细胞L929为对象,从细胞存活率、细胞周期、DNA损伤及活性氧(ROS)的生成方面研究黄连的毒性作用及其可能机制。L929细胞与不同浓度的黄连共培养24 h后,CCK-8法检测细胞存活率,倒置显微镜观察细胞形态变化,流式细胞仪检测细胞周期变化及细胞内ROS的生成情况,彗星实验检测细胞DNA的损伤情况。结果显示,黄连浓度高于1 mg/m L,细胞存活率显著降低,且呈浓度依赖性;高于1 mg/m L的黄连可明显改变细胞的正常形态;2 mg/m L的黄连可使G2/M期细胞比例及细胞内ROS生成显著增多;0.05 mg/m L的黄连可引起DNA各损伤指标明显增加。本实验表明黄连对细胞有毒性作用,其毒性机制与细胞周期阻滞、DNA损伤及细胞内ROS的积累有关。     

PPARγ激动剂15d-PGJ_2对甲状腺刺激抗体作用下人甲状腺细胞存活和凋亡的影响

目的探讨PPARγ激动剂15d-PGJ2对甲状腺刺激抗体(TSAb)作用下人甲状腺细胞存活和凋亡的影响。方法应用原代培养人甲状腺细胞为研究对象,用TSAb刺激细胞24 h,不同浓度15d-PGJ2(0～40μmol/L)干预0～48 h后,MTT法测定细胞存活率;干预24 h后观察细胞凋亡形态、流式细胞技术测定细胞凋亡率。结果 15d-PGJ2呈浓度和时间依赖性抑制TSAb作用下甲状腺细胞的存活(P<0.05);5～40μmol/L15d-PGJ2对TSAb作用下甲状腺细胞有促进凋亡作用(P<0.05),并呈剂量依赖关系;10μmol/L PPARγ拮抗剂GW9662能拮抗20μmol/L15d-PGJ2对TSAb作用下甲状腺细胞的凋亡作用(P<0.05)。结论 15d-PGJ2能够引起TSAb作用下甲状腺细胞凋亡,这一效应可能主要通过PPARγ途径起作用。     

热休克蛋白90在氯化钴对抗血清-葡萄糖剥夺引起的心肌细胞损伤中的作用

目的观察氯化钴(CoCl2)对血清-葡萄糖剥夺(SGD)诱导的H9c2心肌细胞损伤的影响,探讨热休克蛋白90(HSP90)在其中的作用。方法用SGD的方法处理H9c2心肌细胞,建立缺血性损伤的心肌细胞模型。在SGD过程中同时给予CoCl2处理;在应用SGD和CoCl2之前给予HSP90抑制剂(17-AAG)预处理。处理结束后,检测细胞存活率、HSP90的表达、细胞内活性氧(ROS)以及线粒体膜电位(ΔΨm)。结果 SGD处理可引起H9c2心肌细胞损伤,表现为细胞存活率降低、胞内ROS水平升高及ΔΨm丢失。SGD处理还可降低H9c2心肌细胞内HSP90的表达。在50～100μmol.L-1浓度范围内,CoCl2处理可保护H9c2心肌细胞对抗SGD引起的存活率降低,100μmol.L-1CoCl2还可对抗SGD引起的ROS水平升高和ΔΨm丢失。100μmol.L-1CoCl2可时间依赖性地促进胞内HSP90的表达。在50～200μmol.L-1浓度范围内,CoCl2处理可对抗SGD诱导的HSP90表达下调,其中100μmol.L-1的CoCl2具有最强的拮抗作用。17-AAG通过抑制HSP90的作用可明显减弱CoCl2诱导的上述心肌细胞保护作用。结论 CoCl2可保护H9c2心肌细胞对抗SGD引起的损伤,其机制之一与上调HSP90表达有关。     

硫化氢对抗化学性缺氧引起的心肌细胞损伤及其机制

目的观察H2S对氯化钴(CoCl2)诱导的H9C2心肌细胞缺氧性损伤的影响,探讨其作用机制。方法用CoCl2处理H9C2心肌细胞,建立化学性缺氧诱导心肌细胞损伤的实验模型。NaHS(H2S的供体)在CoCl2处理H9C2心肌细胞前30 min加入培养基中,作为预处理。应用CCK-8比色法检测细胞存活率;PI染色流式细胞仪检测细胞凋亡率;Western blot法检测Cleaved Caspase-3的表达;GSSG试剂盒检测GSSG/(GSSG+GSH)的比值;双氯荧光素(DCFH-DA)染色荧光显微镜照相检测细胞内的活性氧(ROS);罗丹明123(Rh123)染色荧光显微镜照相检测线粒体膜电位(MMP)。结果在400~800μmol.L-1浓度范围内,CoCl2处理H9C2心肌细胞36 h,呈剂量依赖性地降低细胞存活率。在600μmol.L-1CoCl2处理H9C2心肌细胞前30 min,应用400μmol.L-1NaHS可明显地抑制CoCl2对细胞的损伤作用,使细胞存活率明显升高,细胞凋亡率及CleavedCaspase-3表达降低;并使H9C2心肌细胞内GSSG/(GSH+GSSG)比值及ROS水平明显降低,同时明显地改善MMP。结论H2S能明显地对抗化学性缺氧诱导的心肌细胞损伤,此保护作用与其降低GSSG/(GSH+GSSG)比值和ROS水平,改善MMP,抑制Caspase-3活化等机制有关。     

在地昔帕明诱导胶质瘤C6细胞凋亡过程中caspase3基因表达和[Ca^2＋]i稳态^1

目的：研究抗抑郁药地昔帕明（Des)对胶质瘤C6细胞的凋亡诱导作用以及对凋亡关键效应分子caspase3和凋亡早期信号[Ca^2 ]i的调控作用。方法：采用流式细胞术（FCM）和凝胶电泳观察Des对C6细胞凋亡的DNA裂解作用,RT－PCR分析caspase3基因的表达以及激光扫描共聚焦显微镜测量单个活细胞[Ca^2 ]i浓度。结果：Des(10,20,40μmol/L)处理C6细胞24h后,FCM图的G1峰左侧出现凋亡特征性亚二倍体细胞峰,凋亡细胞百分率分别为5.2%,21.9%和41.9%。同时,凝胶电泳显示典型的DNA“梯带”.Des 20 μmol/L处理C6细胞24h可明显增强caspase 3基因的表达,而未经Des处理的C6细胞则检测不到caspase 3基因的表达。此外,Des 40μmol/L可使C6细胞[Ca^2 ]i迅速升高并维持超过28min,而钙螯合剂依他酸可显著降低C6细胞[Ca^2 ]i增高幅度,提示Des致C6细胞[Ca^2 ]i增高主要与细胞外钙内流有关。结论：Des诱导C6胶质瘤细胞凋亡可能与caspase 3基因表达的上调以及细胞内钙稳态的失衡有关。     

依达拉奉保护H9c2心肌细胞对抗异丙肾上腺素诱导的氧化应激及内质网应激反应

目的探讨依达拉奉(EDA)能否保护H9c2心肌细胞对抗异丙肾上腺素(ISO)诱导的氧化应激和内质网应激(ERS)。方法用ISO处理H9c2心肌细胞,建立β1肾上腺素受体持续兴奋诱导心肌细胞毒性的体外模型。EDA在ISO处理心肌细胞前1 h加入培养基中作为预处理。CCK-8比色法检测细胞存活率;双氯荧光素(DCFH-DA)染色/荧光显微镜照相检测细胞内活性氧(ROS)的含量;罗丹明123(Rh123)染色/荧光显微镜照相检测线粒体膜电位(MMP);Western blot法检测葡萄糖调节蛋白78(GRP78)的表达。结果 ISO在20～100μmol.L-1浓度范围内处理H9c2心肌细胞48 h,呈剂量依赖性地降低细胞存活率;80μmol.L-1ISO处理H9c2心肌细胞可使细胞内ROS含量明显增多及MMP明显降低;80μmol.L-1 ISO处理H9c2心肌细胞0～24 h,可时间依赖性地上调内质网应激蛋白GRP78的表达,其中12 h达到高峰。分别用10、20和40μmol.L-1 EDA预处理1 h可以减弱80μmol.L-1 ISO处理H9c2心肌细胞48h引起的细胞存活率降低,500、1 000和2 000μmol.L-1氧自由基清除剂NAC分别预处理1 h也可减轻ISO诱导的心肌细胞毒性反应;40μmol.L-1的EDA预处理1 h可明显减轻ISO诱导的胞内ROS堆积及MMP降低,并明显抑制ISO引起的GRP78的表达上调。结论 EDA可保护H9c2心肌细胞对抗ISO诱导的损伤作用,其机制可能与抗氧化及抑制内质网应激反应有关。     

1-磷酸鞘氨醇后适应对H9c2心肌细胞缺氧/复氧损伤的保护作用

目的 研究1-磷酸鞘氨醇（S1P）后适应对H9c2心肌细胞缺氧复氧损伤的保护作用,并探讨其作用机制。方法 将培养的H9c2心肌细胞随机分为5组,即①正常（Con）组;②缺氧/复氧（H/R）组;③S1P低浓度（L）组;④S1P中浓度（M）组;⑤S1P高浓度（H）组。测定各组H9c2心肌细胞的存活率; 收集细胞培养液测定超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量;流式细胞仪检测心肌细胞凋亡百分率;Fura 2-AM标记细胞内游离钙离子,检测荧光强度以反映细胞内游离钙离子浓度的变化;Western Blot法测定保护性蛋白热休克蛋白70 （HSP70）的表达情况。结果 对于H/R 损伤的H9c2心肌细胞,S1P能够提高细胞的存活率,降低细胞内MDA含量及细胞内钙离子浓度,提高细胞内SOD活力,增强抗凋亡蛋白HSP70的表达,且呈一定的浓度依赖性。结论 S1P可以减轻心肌细胞氧化应激损伤,改善心肌细胞活力并减少凋亡。S1P对心肌细胞的保护作用可能是通过减少Ca2 超负荷,增加抗凋亡蛋白HSP70表达来实现的。     

氯化钴对乳鼠心肌细胞凋亡的影响

目的确定萌化钴(CoCl2)对乳鼠心肌细胞凋亡的影响。方法采用达氏修正依氏(DMEM)/F12(1∶1)培养基原代培养乳鼠心肌细胞,3d后分别给予浓度200、400、600μmol/L的CoCl2培养24h,对照组不加CoCl2。锥虫蓝染色检测细胞存活力,Hochest33258荧光染色检测细胞凋亡,用Western Blot检测缺氧诱导因子-1α(HIF-1α)蛋白水平。结果与对照组比较,CoCl2培养组细胞存活率明显下降(P<0.01),凋亡率明显增加(P<0.01),且心肌细胞存活率下降程度及细胞凋亡率睁加程度均随着氯化钴浓度增加而升高。HIF-1α蛋白水平亦随CoCl2浓度升高而增加。结论CoCl2可模拟缺氧诱导乳鼠心肌细胞凋亡。     

Application of flow cytometry in the study of apoptosis in neonatal rat cardiomyocytes

Depending on the concentration, catecholamines activate various intracellular signaling pathways and can induce apoptosis in cardiac myocytes. Although 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine iodide (JC-1) has been previously used to study mitochondria in intact cardiomyocytes, there have been no reports on the detection of apoptosis in neonatal cardiomyocytes in combination with flow cytometry and confocal microscopy. In our study, neonatal rat cardiomyocytes were exposed to norepinephrine (NE) and isoproterenol (ISO) in concentrations of 1 and 10 microM for 48 h. NE concentrations of 1 and 10 microM decreased the number of viable cardiomyocytes by 18% (*p < 0.05) and 24% (**p = 0.01), respectively. ISO in a concentration of 1 microM increased the number of viable cardiomyocytes by 13% while 10 microM decreased the number of viable cardiomyocytes by 43% (***p < 0.001). Apoptotic cells were detected by flow cytometry and confocal microscopy. NE in concentrations of 1 and 10 microM increased the percentage of apoptotic cells by 12.2% and 34.3%, respectively, while ISO alone in a concentration of 10 microM increased the percentage of apoptotic cells by 11.3%. The results demonstrated that these two methods are reliable and suitable for the detection and study of apoptosis in cultures of neonatal cardiomyocytes.     

Effect of different oxygen pressures and N,N′-diphenyl-p-phenylenediamine on adriamycin® toxicity to cultured neonatal rat heart myocytes

The effect of different oxygen pressures and the antioxidant DPPD (N,N′-diphenyl-p-phenylenediamine) on Adriamycin (doxorubicin) cytotoxicity in highly purified cardiac myocytes was investigated to evaluate the involvement of free radicals in the mechanism of toxicity. Adriamycin exposure caused a time-dependent decrease in viability measured as intracellular potassium ion release or lactate dehydrogenase retention. Incubation of myocytes in 16, 172 or 834 μM oxygen during exposure to 200 μM Adriamycin for 6 hr killed 13, 42 and 56% of the cells in the respective cultures. DPPD prolonged viability in the latter two oxygen concentrations and protected against lipid peroxidation measured as production of malondialdehyde and 4-hydroxynonenal. Addition of superoxide dismutase decreased the Adriamycin-induced cell killing to 6% after a 4-hr incubation, as compared to 24% in cultures exposed to Adriamycin only. Adriamycin exposure decreased the concentration of reduced glutathione, and the toxicity of the drug was increased when glutathione reductase was inhibited by the addition of BCNU (1,3-bis-2-chloroethyl-1-nitrosourea). No significant effect on Adriamycin toxicity was observed after inhibition of glutathione synthesis by treatment with BSO (buthionine sulfoximine). It is concluded that free radicals play an important role in Adriamycin toxicity to heart myocytes, and that the cell killing mechanism is likely to be related to induction of lipid peroxidation.     

Protective effect of tubuloside B on TNFalpha-induced apoptosis in neuronal cells.

AIM: To investigate the neuroprotective effect of tubuloside B, one of the phenylethanoids isolated from the stems of Cistanche salsa, on tumor necrosis factor-alpha (TNFalpha)-induced apoptosis in SH-SY5Y neuronal cells. METHODS: Cell viability was analyzed using MTT assay. Apoptotic cells were detected using Hoechst33342 staining, and confirmed by DNA fragmentation and flow cytometric analysis. The activity of caspase-3 was measured with special assay kit. The concentration of free intracellular calcium was determined with the probe Indo-1 by spectrometer. The level of intracellular reactive oxygen species and the potential of mitochondrial membrane were determined by laser scanning confocal microscopy (LSCM) combined with fluorescence probe H2DCFDA or JC-1 respectively. RESULTS: SH-SY5Y cells treated with TNFalpha 100 microg/L for 36 h showed typical morphological changes of apoptosis. DNA ladder could be observed by agarose gel electrophoresis. The highest percentage of apoptotic cells accumulated to 37.5 %. Following 36 h treatment with TNFalpha, accumulation of intracellular ROS and [Ca2+]i and decrease in mitochondrial membrane potential were observed, and caspase-3 activity increased by about five-fold compared with controls. However, pretreatment with tubuloside B (1, 10, or 100 mg/L) for 2 h attenuated the TNFalpha-mediated apoptosis. The antiapoptotic action of tubuloside B was partially dependent on an anti-oxidative stress effects, maintain of mitochondria function, decrease of concentration of free intracellular calcium and inhibition of caspase-3 activity. CONCLUSION: Tubuloside B has the neuroprotective capacity to antagonize TNFalpha-induced apoptosis in SH-SY5Y cells and may be useful in treating some neurodegenerative diseases.     

哌唑嗪对去甲肾上腺素诱导心肌细胞凋亡的影响

目的：研究α受体拮抗剂哌唑嗪对去甲肾上腺素诱导的原代培养心肌细胞凋亡的影响。方法：采用噻唑蓝（MTT）法分析原代培养心肌细胞存活率;乳酸脱氢酶（LDH）外漏实验和吉姆萨染色实验观察原代心肌细胞损伤;JC-1免疫荧光实验检测线粒体膜电位变化;Annexin V-FITC/PI双染,流式细胞仪检测细胞凋亡。结果：NE（100 μmol·L^-1）作用于原代培养心肌细胞48 h,细胞存活率降低;LDH外漏增加;细胞形态改变,细胞数减少、胞体膨大、胞核居中、胞间间隙增大,呈明显病理学特征;线粒体膜电位降低;心肌细胞凋亡率增加。哌唑嗪明显改善去甲肾上腺素诱导的原代培养心肌细胞损伤,增加MTT值、降低LDH外漏量、改善心肌细胞病理形态、增加心肌细胞线粒体膜电位、降低心肌细胞凋亡率（P<0.05或P<0.01）。结论：哌唑嗪能显著抑制去甲肾上腺素诱导的心肌细胞凋亡,本实验为α受体阻断剂改善高交感诱导心肌细胞凋亡提供实验基础与理论指导。     

Toxic Effects of Free and DNA-linked Daunorubicin on Isolated Rat Cardiac Myocytes

Abstract: Isolated cardiac myocytes from adult rats were used as a model to study the cardiotoxicity of free and DNA-linked daunorubicin. The toxic effects on the myocytes were evaluated by studying morphological changes, trypan blue exclusion and cell membrane permeability to NADH, as determined by LDH-activity. At a concentration of 100 μM daunorubicin caused an increased plasma membrane permeability within 30 min. Using light microscopy, the myocytic injury induced by daunorubicin could be distinguished from that induced by anoxia or elevated pH. In contrast to the effect of the free drug, no toxic effects could be demonstrated after incubation with DNA-linked daunorubicin (100 μM) for 5 hours. The higher toxicity of the free drug was related to a much higher intracellular accumulation of daunorubicin. No fluorescent metabolites of daunorubicin could be detected in the myocardial cells. Daunorubicin did not induce lipid peroxidation, as judged by the absence of malondialdehyde production and evolution of ethane. It is concluded that daunorubicin exerts toxic effects on rat cardiac myocytes by mechanisms that do not involve lipid peroxidation. Isolated cardiac cells from adult rats seem to be a useful model for the further study of such mehanisms.     

黄芪多糖对异丙肾上腺素诱导的乳大鼠心肌细胞肥大的保护作用

目的探讨黄芪多糖(APS)对异丙肾上腺素(isoprot-erenol,Iso)诱导的乳鼠心肌细胞肥大的保护作用及机制。方法以原代培养乳鼠心肌细胞为模型,应用Iso 10μmol.L-1诱导心肌细胞肥大,观察不同浓度的黄芪多糖及L型钙通道阻滞剂维拉帕米(Verapamil,VER)对心肌肥厚的影响。用Lowry法测心肌细胞蛋白含量;消化分离法及计算机图像分析系统测细胞体积;RT-PCR法检测心肌细胞ANP的mR-NA表达;以Fluo-3/AM为荧光探针,采用激光共聚焦显微镜观察细胞内[Ca2+]i的变化。结果 APS和VER均能够有效抑制Iso诱导的心肌细胞肥大,表现为蛋白质含量降低,体积减小,ANP mRNA表达降低和细胞内钙离子浓度降低,且APS的作用呈一定的剂量依赖性。结论黄芪多糖对Iso诱导乳大鼠心肌细胞肥大有保护作用,其机制可能与降低[Ca2+]i有关。     

Resveratrol protects against arsenic trioxide-induced cardiotoxicity in vitro and in vivo

BACKGROUND AND PURPOSE: The clinical use of arsenic trioxide (As(2)O(3)), a potent antineoplastic agent, is limited by its severe cardiotoxic effects. QT interval prolongation and apoptosis have been implicated in the cardiotoxicity of As(2)O(3). The present study was designed to evaluate the effects of resveratrol on As(2)O(3)-induced apoptosis and cardiac injury. EXPERIMENTAL APPROACH: In a mouse model of As(2)O(3)-induced cardiomyopathy in vivo, QT intervals and plasma enzyme activities were measured; cardiac tissues were examined histologically and apoptosis assessed. In H9c2 cardiomyocyte cells, viability, apoptosis, generation of reactive oxygen species (ROS) and cellular calcium levels were measured. KEY RESULTS: In the mouse model, resveratrol reduced As(2)O(3)-induced QT interval prolongation and cardiomyocyte injury (apoptosis, myofibrillar loss and vacuolization). In addition, increased lactate dehydrogenase activity and decreased activities of glutathione peroxidase, catalase and superoxide dismutase were observed in the plasma of As(2)O(3)-treated mice; these changes were prevented by pretreatment with resveratrol. In As(2)O(3)-treated H9c2 cardiomyocytes, resveratrol significantly increased cardiomyocyte viability and attenuated cell apoptosis as measured by acridine orange/ethidium bromide staining, TdT-mediated dUTP nick end labelling assay and caspase-3 activity. As(2)O(3)-induced generation of ROS and intracellular calcium mobilization in H9c2 cells was also suppressed by pretreatment with resveratrol. CONCLUSIONS AND IMPLICATIONS: Our results showed that resveratrol significantly attenuated As(2)O(3)-induced QT prolongation, structural abnormalities and oxidative damage in the heart. In H9c2 cardiomyocytes, resveratrol also decreased apoptosis, production of ROS and intracellular calcium mobilization induced by treatment with As(2)O(3). These observations suggested that resveratrol has the potential to protect against cardiotoxicity in As(2)O(3)-exposed patients.