滋膵通脉饮通过调节PI3K/Akt/mTOR信号通路减轻高糖诱导的H9c2心肌细胞损伤的实验研究

doi:10.3969/j.issn.1006-6233.2023.01.006

摘要
图/表
参考文献(15)
相关文章 (15)

全文: PDF (1436 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

摘要 目的: 通过对高糖诱导的H9c2心肌细胞自噬和凋亡可能涉及的信号通路(PI3K-Akt-mTOR)进行研究,探讨滋膵通脉饮抑制心肌细胞凋亡的作用机制。方法: SD大鼠随机分为空白血浆组、滋膵通脉饮组,每组10只。连续灌胃7d,制备相应的含药血浆。滋膵通脉饮组药物浓度为4.4g/mL,灌胃量为10mL·kg^-1·d^-1;空白血浆组采用等剂量的灭菌超纯水灌胃(给药体积为10mL/kg)。把H9c2心肌细胞分为正常组、空白血浆组和含药血浆组,分别加入胎牛血清,空白血浆和含药血浆,每组均设置5%、10%、15%的浓度梯度。通过CCK-8法比较各组H9c2心肌细胞增殖情况,计算细胞存活率,筛选含药血浆。将高糖诱导的H9c2心肌细胞随机分成3 组:模型组、滋膵通脉饮含药血浆组和恩格列净组,加上正常组,共4组,给药24h后,IF检测 LC3B在H9c2心肌细胞的表达。Western blot法检测PI3K-Akt-mTOR信号通路蛋白和凋亡相关蛋白Caspase-3的表达。结果: 含药血浆及空白血浆在10%的药物浓度水平对细胞增殖无明显影响,作为本次含药血浆实验干预的浓度。与正常组比较,模型组心肌细胞LC3B Ⅱ/LC3BⅠ表达明显降低,PI3K、P-Akt、mTOR和Caspase-3的蛋白表达明显升高(P<0.05);与模型组比较,滋膵通脉饮含药血浆组与恩格列净组LC3B Ⅱ/LC3BⅠ表达明显升高,PI3K、P-Akt、mTOR和Caspase的蛋白表达明显降低(P<0.05);滋膵通脉饮含药血浆组与恩格列净组LC3B Ⅱ/LC3BⅠ表达、PI3K、P-Akt、mTOR和Caspase-3的蛋白表达差异无统计学意义(P>0.05)。结论: 滋膵通脉饮可激活心肌细胞自噬而抑制心肌细胞凋亡,这可能与抑制PI3K/P-Akt/mTOR信号通路有关。

	服务

	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章

关键词 ：滋膵通脉饮, H9c2心肌细胞, 自噬, 凋亡, PI3K-Akt-mTOR

Abstract：Objective: To investigate the possible signaling pathways (PI3K-Akt-mTOR) involved in autophagy and apoptosis of H9c2 cardiomyocytes induced by high glucose, and to investigate the mechanism of action of Zicui Tongmai Decoction in inhibiting cardiomyocyte apoptosis. Methods: SD rats were randomly divided into blank plasma group and Zicui Tongmai Decoction group, with 10 rats in each group. The corresponding medicated plasma was prepared by gavage for 7 days,the drug concentration of Zicui Tongmai Decoction group was 4.4g/mL and the gavage volume was 10mL·kg^-1·d^-1,the blank plasma group was gavaged with the same dose of sterilized ultrapure water (the administration volume was 10mL/kg). H9c2 myocardial cells were divided into normal group, blank plasma group and drug containing plasma group, fetal bovine serum, blank plasma and drug containing plasma were added respectively. A concentration gradient of 5%, 10% and 15% was set for each group. The proliferation of H9c2 cardiomyocytes in each group was compared by CCK-8 method, the cell survival rate was calculated, and the drug containing plasma was screened. H9c2 cardiomyocytes induced by high glucose were randomly divided into three groups: model group, Zicui Tongmai Decoction medicated plasma group and engliejing group, and additionally there was normal group. Expression of LC3B in H9c2 cardiomyocytes was detected 24h after administration of IF, and the expression of PI3K Akt mTOR signal pathway protein and apoptosis related protein caspase-3 was detected by Western blot. Results: Drug containing plasma and blank plasma have no significant effect on cell proliferation at the concentration level of 10%, which is used as the intervention concentration of drug containing plasma experiment. The expression of LC3B Ⅱ/LC3BⅠ decreased significantly compared with the normal group(P<0.05),but the protein expression of PI3K, P-Akt, mTOR and Caspase-3 increased compared with the normal group(P<0.05). The expression of LC3B Ⅱ/LC3BⅠin Zicui Tongmai Decoction medicated plasma group and engliejing group increased significantly compared with the model group(P<0.05),but the protein expression of PI3K,P-Akt,mTOR and Caspase-3 in Zicui Tongmai Decoction medicated plasma group and engliejing group decreased compared with the model group(P<0.05). There was no significant difference in expression of LC3BⅡ/LC3BⅠ and protein expression of PI3K,P-Akt,mTOR and Caspase-3 between Zicui Tongmai Decoction medicated plasma group and engliejing group (P>0.05). Conclusion: Zicui Tongmai Decoction can activate autophagy and inhibit apoptosis of cardiomyocytes, which may be related to the inhibition of PI3K/p-Akt / mTOR signaling pathway.

Key words： Zicui Tongmai decoction H9c2 cardiomyocytes Autophagy Apoptosis PI3K-Akt-mTOR

基金资助:湖南省中医药管理局一般项目,(编号:2021071);湖南省长沙市自然科学基金项目,(编号:Kq2202476)

通讯作者: 卜献春

引用本文:

吴刚强, 袁春云, 袁思斯, 毛叶, 陈琪, 温小凤, 谭军, 卜献春, 李非, 李洁花. 滋膵通脉饮通过调节PI3K/Akt/mTOR信号通路减轻高糖诱导的H9c2心肌细胞损伤的实验研究[J]. 河北医学, 2023, 29(1): 31-36.
WU Gangqiang, YUAN Chunyun, YUAN Sisi, et al. Experimental Study on Reducing High Glucose-Induced Injury in H9c2 Cardiomyocytes by Modulating PI3K/Akt/mTOR Signaling Pathway with Zicui Tongmai Decoction. HeBei Med, 2023, 29(1): 31-36.

链接本文:

http://www.hbyxzzs.cn/CN/10.3969/j.issn.1006-6233.2023.01.006 或 http://www.hbyxzzs.cn/CN/Y2023/V29/I1/31

[1] 尧青,李勇,陈晓,等.姜黄素对糖尿病大鼠心肌自噬、凋亡及AMPK-mTOR信号通路的影响[J].中国医院药学杂志,2022,42(10):1004-1008.
[2] 杜琎,石开虎,赵扬,等.丹参酚酸B通过抑制PI3K/Akt/mTOR通路促进自噬减轻大鼠心肌纤维化的研究.[J].现代生物医学进展,2019,19(20):3812-3817.
[3] 吴刚强,熊春红,毛叶,等.滋膵通脉饮对糖尿病心肌病大鼠心肌纤维化和转化生长因子β1/Smads3信号通路的影响[J].中药新药与临床药理,2021,32(1):29-35.
[4] 吴刚强,毛叶,温小凤,等.滋膵通脉饮对糖尿病心肌病大鼠心肌重构的影响[J].中西医结合心脑血管病杂志,2021,19(2):240-243.
[5] Wang Y,Liang B,Lau WB,et al.Restoring diabetes-induced autophagic flux arrest in ischemic/reperfused heart by ADIPOR(adiponectinreceptor)activation involves both AMPK-dependent and AMPK-independent signaling[J].Autophagy,2017,13(11):1855 -1869.
[6] Li Xuefei,Hu Xiaorong,Wang Jichun,et al.Inhibition of autophagy via activation of PI3K/ Akt/ mTOR pathway contributes to the protection of hesperidin against myocardial ischemia/ reperfusio injury[J].International Journal of Molecular Medicine,2018,42(4):1917-1924.
[7] 王晓平,张雪峰,邵明燕,等.自噬信号通路在心力衰竭中的作用以及中药干预的研究进展[J].中华中医药杂志,2019,34(9):4220-4223.
[8] Fan C,Tang X,Ye M,et al.Qi-Li-Qiang-Xin aleviates isoproterenol-induced myocardial injury by inhibiting excessive autophagy activating AKT/mTOR pathway[J].Front Pharmacol,2019,10:1-4.
[9] 饶小娟,吴毓敏,王彦.麦冬皂苷介导AMPK/Akt/mTOR通路对糖尿病大鼠心肌细胞自噬的影响[J].中国循证心血管医学杂志,2018,10(11):1362-1367.
[10] 诺明,陶慧,哈森塔娜,等.舒芬太尼对心肌缺血/再灌注大鼠心肌细胞凋亡及Bcl-2、LC3Ⅱ、Beclin-1表达的影响[J].中西医结合心脑血管病杂志,2020,18(1):50-54.
[11] 殿臣,宫丽鸿,肖福龙.稳斑汤含药血清对巨噬细胞泡沫化凋亡相关蛋白Bax、Bcl2和caspase3表达的影响[J].辽宁中医杂志,2018,45(6):1309-1312.
[12] Liu X,Tan W,Yang F,et al.Shengmai injection reduces apoptosis and enhances angiogenesis after myocardial ischaemia and reperfusion injury in rats[J].Biomed Pharmacother,2018,104(8):629-636.
[13] 田瑛,邓小敏,苏冬,等.基于网络药理学探讨增液汤治疗糖尿病的作用机制[J].广西医学,2021,43(15):1868-1872.
[14] 杨嫒萍,张磊,李炜,等.黄芪甲苷对糖尿病心肌病小鼠的治疗作用及其机制[J].山东医药,2022,62(23):19-24.
[15] 陈梓健,蒋正午,高乐,等.地龙提取物对2型糖尿病心肌病的治疗作用与机制探讨[J].局解手术学杂志,2020,29(8):616-619.