番茄红素对血管紧张素Ⅱ诱导H9c2心肌细胞凋亡的影响

摘要

目的 研究番茄红素(Lyc)对血管紧张素Ⅱ(AngⅡ)诱导H9c2心肌细胞凋亡的保护作用,并探讨其可能的机制.方法 采用AngⅡ(10 μmol/L)刺激H9c2细胞,观察不同浓度Lyc对AngⅡ诱导细胞凋亡的保护效果.采用CCK8检测不同浓度Lyc和(或)AngⅡ对H9c2细胞活性的影响;采用实时半定量逆转录聚合酶链反应(RT-PCR)检测细胞中Bax、Bcl-2的基因表达水平;免疫印迹(Western blot)法检测细胞中Bax、Caspase3、Caspase9、Bcl-2的蛋白表达量;TUNEL染色观察细胞的凋亡比率.结果 和对照组相比,AngⅡ组中H9c2细胞活性为(92.87±4.37)％,当Lyc与AngⅡ同时干预H9c2细胞后,细胞活性显著上升(P＜0.05);RT-PCR结果显示AngⅡ可上调H9c2胞内Bax mRNA表达,Bcl-2表达下调;当AngⅡ与Lyc同时作用时,Bax表达下调而Bcl-2表达上调,且这种改变具有浓度依赖性;Bax、Caspase3、Caspase9在AngⅡ的作用下蛋白表达量上调,Bcl-2下调(P＜0.05),而AngⅡ与Lyc同时作用时可逆转这一现象.TUNEL染色结果显示,AngⅡ作用于H9c2细胞时,可见大量凋亡细胞;而Lyc与AngⅡ同时作用时,凋亡细胞的数量明显减少(P＜0.05).结论 Lyc能够明显改善AngⅡ诱导的H9c2心肌细胞凋亡,提示其对于改善临床上各种心血管疾病可能有着重要的作用.%Objective To investigate the effect of lycopene (Lyc) on H9c2 cell apoptosis induced by angiotensin Ⅱ (Ang Ⅱ).Methods Using Ang Ⅱ (10 μmol/L) to stimulate H9c2 cells,we observed the protective effect of Lyc on H9c2 cells apoptosis.The H9c2 cells viability induced by different consideration of Lyc or Ang Ⅱ or both was detected by CCK8 assay.The expression levels of Bax and Bcl-2 in H9c2 cells were determined by real-time quantitative reverse transcription polymerase chain reaction (RT-PCR).Western blot was conducted to detect the protein expressions of Bax,Caspase 3,Caspase 9 and Bcl-2 in H9c2 cells.The apoptotic ratio of H9c2 cells was observed by TUNEL assay.Results Compared with control group,Ang Ⅱ could decrease the viability of H9c2 cells to (92.87±4.37)％.The result of RT-PCR showed that Ang Ⅱ decreased the expression level of Bcl-2,and Bax level was increased under the stimulation of Ang Ⅱ (P＜0.05),while the expression level of Bcl-2 was increased and Bax level was decreased under the co-stimulation of Ang Ⅱ and Lyc in a concentration dependent manner,which indicated that Lyc ameliorated the apoptosis of H9c2 cells.The result of western blot showed that the protein expressions of Bax,Caspase 3 and Caspase 9 were increased,but Bcl-2 was decreased after the stimulation of Ang Ⅱ (P＜0.05).While these phenomenon reversed apparently under the co stimulation of Ang Ⅱ and Lyc.A large number of apoptotic cells were observed under the stimulation of Ang Ⅱ through TUNEL assay.But the number of apoptotic cells reduced significantly under the co-stimulation of Lyc and Ang Ⅱ (P ＜0.05).Conclusions Lyc ameliorates the H9c2 cell apoptosis induced by Ang Ⅱ,which indicates that Lyc may have an important role in the treatment of various cardiovascular diseases.