丹参素对缺氧/复氧诱导H9C2心肌细胞损伤和内质网应激的改善作用

目的观察丹参素(DLA)通过抑制内质网应激和凋亡对H9C2心肌细胞缺氧/复氧(H/R)后损伤的保护作用。方法将离体培养的H9C2心肌细胞随机分为3组:对照组、缺氧/复氧组(H/R组)、DLA+H/R组(DLA组)。H/R组先缺氧6h后复氧24h;DLA组于缺氧时给予DLA(10-6M)培养6h,再复氧24h。对照组心肌细胞正常培养至实验结束。ELISA测定细胞中乳酸脱氢酶(LDH)、肌酸激酶MB同工酶(CK-MB)和丙二醛(MDA)的含量;Western blot检测细胞中内质网应激蛋白葡萄糖调节蛋白78(GRP78)、Caspase-12、Bax、Bcl-2和SOD的蛋白含量。结果与对照组比较,H/R组细胞LDH、CK-MB和MDA的含量显著升高(P<0.05),GRP78、Caspase-12和Bax蛋白表达明显增加(P<0.05),而Bcl-2和SOD的蛋白含量明显降低(P<0.05);给予DLA可明显改善上述指标的变化。结论H/R可诱导H9C2心肌细胞发生凋亡、氧化应激和内质网应激,引起细胞损伤和凋亡;DLA可以通过抑制凋亡、氧化应激和内质网应激,减少细胞损伤,发挥保护细胞的作用。

Ameliorating effects of Danshensu on hypoxia/reoxygenation-induced H9C2 cardiomyocytes injury and endoplasmic reticulum

Objective To observe the protective effect of DLA on hypoxia/reoxygenation(H/R)injury by attenuating endoplasmic reticulum stress(ERS)and apoptosis in H9C2 cardiomyocytes.Methods The H9C2 cardiomyocytes were randomly divided into three groups:control group,hypoxia/reoxygenation group(H/R group),DLA+H/R group(DLA group).H/R group underwent hypoxia for 6 hfollowed by reoxygenation for 24 h.DLA group was given at the time of hypoxia for 6 h,then reoxygenation for 24 h.H9C2 cardiomyocytes of control group were cultured under normal condition till the end of experiment.Lactic dehydrogenase(LDH),CK-MB,MDA and SOD activity were measured by ELISA and the protein expression of glucose-regulated proteins 78(GRP78),Caspase-12,Bcl-2,Bax and SOD were detected by western blot.Results Compared with the control group,increased levels of LDH、CK-MB and MDA,increased expressions of GRP78,Caspase-12 and Bax protein,decreased activity of SOD,decreased expression of Bcl-2 and SOD protein were shown in the H/R group significantly,while DLA ameliorated the above indicators.Conclusion H/R can induce ERS leading to cell injury and apoptosis.DLA can protect H9C2 against H/R injury by inhibiting ERS and apoptosis.