红景天苷可减少高糖诱导的心肌细胞炎症和凋亡

Salidroside protects cardiomyocytes against high glucose-induced inflammation and apoptosis

目的:探讨和了解红景天苷(SAL)对高糖培养的心肌细胞(CM)炎症和凋亡的抑制作用。方法:H9c2细胞培养分为5组,对照组(C)、高糖组(HG)、SAL干预组(SAL)、JNK抑制剂SP600125干预组(SP)和SAL+SP组。采用CCK8试剂盒检测细胞活性;各组H9c2细胞均培养96 h后,分别采用ROS试剂盒检测细胞内ROS水平、ELISA方法检测细胞培养上清液中IL-6和TNF-α表达水平和Western blot方法检测H9c2细胞内Bcl-2、Bax、JNK和p-JNK蛋白表达水平。结果:根据CCK8方法检测细胞活性的结果分别确定高糖诱导时间为96 h和SAL有效浓度为10μmo/l L。SAL组以及SAL+SP组检测结果显示,均可有效抑制ROS产生(P<0.05)。单独SP组对ROS无显著抑制作用(P>0.05)。SAL组可下调高糖诱导的p-JNK表达(P<0.05)。SP和SAL+SP组能抑制高糖诱导的JNK和p-JNK表达(P<0.05)。SAL、SP以及SAL+SP这三组均能有效降低高糖诱导的H9c2细胞IL-6和TNF-α的表达(P<0.05)。SAL组和SP组不能抑制高糖诱导的Bcl-2表达下调(P>0.05),但能下调Bax表达水平并提高Bcl-2/Bax值(P<0.05)。SAL+SP组可有效增加高糖诱导的Bcl-2表达(P<0.05),同时显著下调Bax表达以及显著增加Bcl-2/Bax值(P<0.01)。结论:SAL通过抑制高糖诱导的CM产生ROS,减少JNK信号通路激活,降低CM产生IL-6和TNF-α以及细胞凋亡。SAL具有高糖条件下的CM保护作用,可能与其抗炎抗氧化特性有关,有待今后DM动物实验中进一步证实其对CM的治疗作用。

Objective:The aim of this study was to investigate the effect of salidroside(SAL)on inhibiting cardiomyocytes(CM)from high-glucose inducing inflammation and apoptosis.Methods:The H9c2 cell lines in DMEM cell culture medium were divided into five groups including control group(C),high glucose group(HG),salidroside group(SAL),JNK inhibitor of SP600125 group(SP),SAL+SP group.The H9c2 cell viability was detected by CCK8 method.H9c2 cells were cultured for 96 h.After that,intracellular ROS production was measured by using ROS kits.The IL-6 and TNF-αexpression levels were detected by ELISA method in cell culture supernatant,and intracellular protein expression levels of Bcl-2/Bax,JNK/p-JNK in H9c2 cells were detected by Western blot.Results:According to the results of CCK8 assay,the high glucose induction time was 96 h and the effective concentration of SAL was 10μmo/l L.The results of SAL group and SAL+SP intervention group showed that ROS production could be effectively inhibited(P<0.05).The SP group alone had no significant inhibitory effect on ROS(P>0.05).The SAL group could downregulate the expression of p-JNK induced by high glucose(P<0.05).The SP and the SAL+SP group could inhibit the expression of JNK and p-JNK induced by high glucose(P<0.05).The expression of IL-6 and TNF-αin H9c2 cells induced by high glucose was significantly decreased in SAL group,SP group and SAL+SP group(P<0.05).SAL and SP groups could not inhibit the downregulation of Bcl-2 expression induced by high glucose(P>0.05),but could decrease the expression level of Bax and increase the ratio of Bcl-2/Bax(P<0.05).SAL+SP combined with the intervention group could effectively increase the expression of Bcl-2 induced by high glucose(P<0.05),significantly down-regulate the expression of Bax and significantly increase the ratio of Bcl-2/Bax(P<0.01).Conclusion:The salidroside inhibited the production of ROS by CM induced by high glucose,prevented the activation of JNK signaling pathway,and reduced the production of IL-6 and TNF-αby CM as well as cell apoptosis.SAL has protective effect on cardiomyocytes in response to high glucose circumstances.Probably,these findings are related to anti-inflammatory and anti-oxidant properties of SAL.However,performing further diabetic animal studies are necessary to support the therapeutic effects of SAL in cardiac dysfunction.