| 张建成,林淑琴,胡鹏飞.丹参多糖促进自噬抑制脂多糖诱导的心肌细胞凋亡的实验研究[J].浙江中西医结合杂志,2020,30(4): |
| 丹参多糖促进自噬抑制脂多糖诱导的心肌细胞凋亡的实验研究 |
| SMP protects myocardial cells against LPS-induced apoptosis through promoting autophagy |
| 投稿时间：2019-10-03 修订日期：2020-03-06 |
| DOI： |
| 中文关键词: 丹参多糖 心肌细胞 凋亡 自噬 |
| 英文关键词:SSalvia miltiorrhiza Polysaccharide Cardiomyocytes Apoptosis Autophagy |
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| 中文摘要: |
| 目的 探究丹参多糖(SMP)调控脂多糖(LPS)诱导的心肌细胞凋亡的机制。方法 不同浓度(0.0.5、1、2mg/mL)SMP预处理乳鼠心肌细胞12h后,1μg/mL LPS刺激乳鼠心肌细胞24h,以LPS单独处理构建心肌细胞损伤模型,对照组加入等量生理盐水,四唑盐比色法(MTT)观察细胞存活率改变,流式细胞仪检测细胞凋亡比率改变,免疫印迹(Western blot)检测各组细胞自噬相关蛋白LC3、Beclin1及凋亡相关蛋白Bax、Caspase3变化；SMP联合自噬抑制剂氯喹(CQ)或自噬激活剂雷帕霉素靶蛋白抑制剂(Torin1)处理LPS处理的心肌细胞,MTT观察细胞存活率改变,流式细胞仪(FACS)检测细胞凋亡比率改变,Western blot检测各组细胞自噬相关蛋白LC3、Beclin1及凋亡相关蛋白Bax、Caspase3变化。结果 相比于对照组,LPS 处理乳鼠心肌细胞24h,细胞活力显著下降,凋亡细胞比例升高,凋亡相关蛋白Bax、cleaved-Caspase3表达增多,自噬相关蛋白Beclin1和LC3II表达增多。相比于LPS组,丹参多糖处理组细胞活力升高,凋亡细胞比例下降,Bax、cleaved-Caspase3蛋白表达减少, LC3-II 、Beclin1蛋白表达增多；相比于丹参多糖处理组,CQ抑制细胞活力,增加细胞凋亡数,促进Bax、cleaved-Caspase3和LC3II蛋白表达,抑制Beclin1蛋白表达,Torin1上调细胞活力,减少细胞凋亡数,促进LC3-II 、Beclin1蛋白表达,抑制Bax、cleaved-Caspase3蛋白表达。结论 SMP促进自噬抑制LPS诱导的心肌细胞自噬凋亡。 |
| 英文摘要: |
| Objective To investigate the protective effect of Salvia miltiorrhiza Polysaccharide (SMP) on lipopolysaccharide (LPS)-induced cardiomyocytes and explore its mechanism. Methods Cardiomyocytes were treated with SMP at different concentrations (0.5, 1, 2 mg/mL) for 12h before stimulating with 1 μg/mL LPS for 24 h. Cell viability was observed by MTT, the apoptosis rate was detected by FACS and the expressions of LC3, Beclin1, Bax and Caspase3 were detected by Western blot; Cardiomyocytes were treated with SMP and CQ/Torin1 for 12h before stimulating with LPS. Cell viability was observed by MTT, the apoptosis rate was detected by FACS and the expressions of LC3, Beclin1, Bax and Caspase3 were detected by Western blot. Results Compared with the control group, the cell viability of the cardiomyocytes treated with LPS was significantly decreased, the proportion of apoptotic cells, the expression of Bax, Beclin1, LC3II and cleaved-Caspase3 increased; Compared with the LPS group, the cell viability, the expression of Beclin1 and LC3II of SMP+LPS group increased, while the proportion of apoptotic cells, and the expression of Bax and cleaved-Caspase3 decreased; Compared with the SMP+LPS group, the cell viability, the expression of Beclin1 and LC3II of SMP+LPS+Torin1 group increased, while the proportion of apoptotic cells, and the expression of Bax and cleaved-Caspase3 decreased; Compared with the SMP+LPS group, the cell viability, the expression of Beclin1 of SMP+LPS+Torin1 group decreased, while the proportion of apoptotic cells, and the expression of Bax,cleaved-Caspase3 and LC3II increased. Conclusion SMP protects myocardial cells against LPS-induced apoptosis through promoting autophagy. |
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