川芎嗪对缺氧复氧诱导H9C2细胞损伤及凋亡的影响及其机制研究

Study on Effect of Tetramethylpyrazine on Injury and Apoptosis of H9C2 Cells Induced by Hypoxia-reoxygenation and Its Mechanism

目的:探讨川芎嗪在缺氧复氧诱导H9C2细胞损伤中的保护作用。方法:体外培养H9C2心肌细胞,构建心肌细胞缺氧复氧损伤及川芎嗪预处理模型。细胞随机分为5组:对照组(NC组)、缺氧复氧损伤组(HR组)、川芎嗪预处理L组(TMP-L组,60μg/mL),M组(TMP-M组,200μg/mL)和H组(TMP-H组,800μg/mL)。心肌细胞损伤程度以心肌细胞活力(OD值)、乳酸脱氢酶(LDH)活性、超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量表示;细胞凋亡程度通过细胞凋亡率测定,并检测bcl-2和bax基因的表达。结果:与NC组比较,HR组细胞活力显著减弱,LDH释放量显著增加,SOD活性降低,MDA含量增多,出现大量心肌细胞凋亡,bcl-2 mRNA表达显著下降,bax mRNA表达显著升高,差异均有统计学意义(P <0.05);与HR组比较,TMP-M组和TMP-H组细胞活力显著增强,LDH释放量显著减少;SOD活性增强,MDA含量下降;TMP-L组、TMP-M组和TMP-H组心肌细胞凋亡率及bcl-2 mRNA表达显著下降,bax mRNA表达显著升高,差异均有统计学意义(P <0.05)。结论:川芎嗪预处理能减轻心肌细胞的缺氧复氧损伤,其作用可能与提高心肌细胞活力、增强SOD活性、降低LDH释放量、减少MDA生成、抑制心肌细胞凋亡有关。

Objective： To discuss the protective effect of tetramethylpyrazine on injury of H9C2 cells induced by hypoxia-reoxygenation. Methods- Established the model of myocardial cells with injury induced by hypoxia-reoxygenation and pretreatment of tetramethylpyrazine by culturing H9C2 myocardial cells in vitro. The cells were divided into 5 groups randomly, including the control group（NC group）, the hypoxia-reoxygenation（HR group）, the tetramethylpyrazine pretreatment group L （TMP-L group, 60 μg/mL）, group M（TMP-M group, 200 μg/mL） and group H（TMP-H group, 800 μg/mL）. The injury degree of myocardial cells was indicated by such indexes as viability of myocardial cells（OD value）, activity of lactate dehydrogenase （LDH）, activity of superoxide dismutase（SOD） and content of malondialdehyde（MDA）; the apoptosis degree was evaluated by the apoptosis rate, and the expression of bcl-2 and bax gene was detected. Results： Comparing with the NC group, the viability of cells in the HR group was significantly reduced, the release amount of LDH was significantly increased, the activity of SOD was decreased, the content of MDA was increased, the apoptosis of numerous myocardial cells occurred, the expression of bcl-2 mRNA was significantly decreased, and the expression of bax mRNA was significantly increased, differences being significant （P 〈 0.05）. Comparing with the HR group, the viability of cells in the TMP-M group and the TMP-H group was significantly enhanced, the release amount of LDH was significantly decreased; the activity of SOD was increased, the content of MDA was decreased; the apoptosis rate and the expression of bcl-2 mRNA of myocardial cells in the TMP-L group, the TMP-M group and the TMP-H group was significantly decreased, and the expression of bax mRNA was significantly increased, differences being significant （P 〈 0.05）. Conclusion： Pretreatment of tetramethylpyrazine can alleviate the hypoxia-reoxygenation injury of myocardial cells, and its effect may be related to improving the viability of myocardial cells and activity of SOD, reducing the release amount of LDH, decreasing the formation of MDA and inhibiting the apoptosis of myocardial cells.