[1]李兵.丹参多酚酸盐对H2O2诱导心肌细胞凋亡与自噬的影响及机制研究[J].西部中医药,2022,35(10):29-34.[doi:10.12174/j.issn.2096-9600.2022.10.07]
　LI Bing.Influence of SAL on H2O2-Induced Apoptosis and Autophagy of H9c2 Myocardial Cells and the Mechanism Study[J].Western Journal of Traditional Chinese Medicine,2022,35(10):29-34.[doi:10.12174/j.issn.2096-9600.2022.10.07]
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丹参多酚酸盐对H2O2诱导心肌细胞凋亡与自噬的影响及机制研究
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《西部中医药》[ISSN:2096-9600/CN:62-1204/R]

卷:
35
期数:
2022年10期
页码:
29-34
栏目:
出版日期:
2022-10-15

文章信息/Info

Title:
Influence of SAL on H2O2-Induced Apoptosis and Autophagy of H9c2 Myocardial Cells and the Mechanism Study
作者:
李兵
邯郸市中心医院，河北 邯郸 056001
Author(s):
LI Bing
Handan Central Hospital, Handan 056001, China
关键词:
丹参多酚酸盐; 心肌细胞; 凋亡; 自噬; Akt/mTOR/Beclin1通路
Keywords:
SAL; myocardial cells; apoptosis; autophagy; Akt/mTOR/Beclin1 pathway
分类号:
R542.2
DOI:
10.12174/j.issn.2096-9600.2022.10.07
文献标志码:
A
摘要:
目的探讨丹参多酚酸盐（salvianolate，SAL）对过氧化氢（hydrogen peroxide，H2O2）诱导H9c2心肌细胞凋亡与自噬的影响及其机制。 方法以100 μmol/L的H2O2干预对数生长期H9c2细胞4 h，建立心肌细胞氧化应激损伤模型，设H2O2组，SAL50、100、200 mg/L组，另取对数生长期H9c2细胞设为正常组。药物干预24 h后，CCK-8法检测细胞增殖率，流式细胞术检测细胞凋亡，GFP-LC3质粒转染后观察自噬小体，Western blot法检测H9c2细胞蛋白激酶B（protein kinase B，Akt）、磷酸化Akt （phosphorylation Akt，p-Akt）、半胱氨酸蛋白酶3（cysteinyl aspartate specific proteinase 3，Caspase-3）、B细胞淋巴瘤2（B cell lymphoma-2，Bcl-2）、Bcl-2相关X蛋白（Bcl-2 related X protein，Bax）、磷酸化哺乳动物雷帕霉素靶蛋白（phosphorylation mammalian target of rapamycin，p-mTOR）、Beclin1、微管相关蛋白1轻链3（microtubule-associated protein 1 light chain 3，LC3）、P62蛋白表达。 结果与H2O2组比较，SAL 100、200 μg/mL组H9c2细胞增值率显著升高而凋亡率显著降低（P＜0.01），细胞内自噬小体数量减少，p-Akt、p-mTOR、bcl-2表达明显上调而Caspase-3、Bax、Beclin1、LC3-I、LC3-Ⅱ、P62表达明显下调（P＜0.05或P＜0.01），p-Akt/Akt、bcl-2/Bax比值升高而LC3-Ⅱ/LC3-I比值下降（P＜0.01）。 结论SAL对H2O2所致H9c2心肌细胞凋亡与自噬具有抑制作用，其机制可能与激活Akt/mTOR/Beclin1通路而抑制促凋亡、促自噬相关蛋白表达和活化有关。
Abstract:
ObjectiveTo discuss the effects of SAL on the apoptosis and autophagy of H9c2 myocardial cells induced by H2O2 and its mechanism. MethodsH2O2, 100 μmol/L, was used to intervene H9c2 cells in logari-thmic growth phase for four hours, to establish oxidative stress injury models of cardiomyocytes, and the models were divided into H2O2 group, SAL group of 50, 100 and 200 mg/L, besides, H9c2 cells in logarithmic growth phase were selected as the normal group. After drug intervention for 24 hours, CCK-8 method was used to detect cell proliferation rate, flow cytometry to measure cellular apoptosis, to observe autophagosome after GFP-LC3 plasmid transinfection, Western blot method to measure the expressions of Akt, p-Akt, Caspase-3, Bcl-2, Bax, p-mTOR, Beclin1, LC3 and P62 protein in H9c2 cells. ResultsCompared with H2O2 group, the proliferation rate of H9c2 cells was increased while the apoptosis rate was significantly reduced in SAL groups of 100 and 200 μg/mL (P<0.01), the number of autophagosome was lowered in the cells, the expressions of p-Akt, p-mTOR and Bcl-2 were up-regulated apparently while the expressions of Caspase-3, Bax, Beclin1, LC3-1, LC3-Ⅱ and P62 were down regulated remarkably (P<0.05 or P<0.01), the ratios of p-Akt/Akt, Bcl-2/Bax were increased while LC3-Ⅱ/LC3-I ratio was decreased (P<0.01). ConclusionSAL owns inhibitory effects on the apoptosis and autophagy of H9c2 myocardial cells induced by H2O2, and its mechanism might be related to the activation of Akt/mTOR/Beclin1 pathway, and inhibiting the expressions and activation of pro-apoptosis and pro-autophagy related proteins.

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备注/Memo

备注/Memo:
李兵（1978—），男，硕士学位，副主任医师。研究方向：急诊医学。河北省医学科学研究课题（20191845）。
更新日期/Last Update: 2022-10-15