丹参多酚酸盐对过氧化氢所致乳鼠心肌细胞凋亡的影响

Effects of Salvianolate on neonatal rat cadiocytes apoptosis induced by H2O2

关键词:  丹参多酚酸盐  过氧化氢  心肌细胞  凋亡  影响

Key words:  Salvianolate  H₂O₂  cardiomyocyte  apoptosis  effect

摘要:

[目的] 研究丹参多酚酸盐（Salvianolate）对过氧化氢（H₂O₂）所致乳鼠心肌细胞凋亡的影响，并探讨其可能的作用机制。[方法] 于无菌环境下分离并培养乳鼠心肌细胞72 h后随机分为空白对照组、H₂O₂组、丹参多酚酸盐（50、100和200 mg/L）+H₂O₂组，每组设8个复孔；给药干预24 h后，检测培养液中谷草转氨酶（AST）、磷酸肌酸激酶（CPK）、乳酸脱氢酶（LDH）含量，四甲基噻唑蓝（MTT）法检测细胞存活率，通过流氏细胞术（Flow Cytometry）检测细胞凋亡状况并计算凋亡率；逆转录-聚合酶链反应（RT-PCR）法检测AKT mRNA和bcl-2 mRNA、Bax mRNA表达并计算bcl-2/Bax表达比值，Western blotting法检测细胞中半胱氨酸天冬氨酸蛋白酶-3（caspase-3）、核转录因子-κB（NF-κB）表达并进行半定量分析，测定细胞中超氧化物歧化酶（SOD）、过氧化氢酶（CAT）活性和丙二醛（MDA）含量。[结果] 与H₂O₂组比较，丹参多酚酸盐（100、200 mg/L）+H₂O₂组培养液中AST、CPK、LDH含量显著降低，细胞存活率显著升高，细胞凋亡率显著降低。细胞中细胞中AKT mRNA、bcl-2 mRNA表达显著上调，Bax mRNA表达显著下调，bcl-2/Bax表达比值显著升高，caspase-3、NF-κB表达显著下调，SOD、CAT活性显著升高且MDA含量显著降低，上述差异均具有统计学意义（P<0.05或P<0.01）。[结论] 丹参多酚酸盐对H₂O₂所致乳鼠心肌细胞凋亡具有抑制作用。

Abstract:

[Objective] To investigate the effects of Salvianolate on neonatal rat cadiocytes apoptosis induced by H₂O₂. [Methods] Cadiocytes of neonate rat was cultivated for 72 hours and divided into six groups:normal control group, H₂O₂ group, Salvianolate (50, 100, 200 mg/L)+H₂O₂ groups(n=8). The 12 hours after the drugs were given. The content of AST, CPK, LDH in culture medium were detected; the survival rate was detected by MTT; the cardiomyocytes apoptosis was observed by flow cytometry and the apoptosis rate was calculated; the expression of AKT mRNA, bcl-2 mRNA, Bax mRNA were detected by RT-PCR and the ratio of bcl-2/Bax was calculated; the expression of caspase-3, NF-κB protein was detected by Western blot and Semi-quantitative analysized; the activity of SOD, CAT and the content of MDA in cardiomyocytes were determinted. [Results] Compared with the H₂O₂ group, the content of AST, CPK, LDH in culture medium of Salvianolate (100, 200 mg/L)+H₂O₂ groups were significantly decreased; the survival rate were significantly increased; the cardiomyocytes apoptosis were improved and the apoptosis rate were significantly decreased; the expression of AKT, bcl-2 mRNA were significantly up-regulated, while the expression of Bax mRNA was significantly down-regulated, the ratio of bcl-2/Bax were significantly increased; the expression of caspase-3, NF-κB protein were significantly down-regulated; the activity of SOD, CAT in cardiomyocytes were significantly increased and the content of MDA was significantly decreased, all of the difference above were significant (P<0.05 or P<0.01). [Conclusion] Salvianolate had inhibitive effects on neonatal rat cadiocytes apoptosis induced by H₂O₂.