玉郎伞黄酮通过AMPK/mTOR信号通路调控自噬对心肌细胞肥大的保护作用OA
Effect of Yulangsan chalcone on autophagy intervention in cardiomyocyte hypertrophy through AMPK/mTOR signaling
目的 基于 AMPK/mTOR 信号通路调控自噬,探讨玉郎伞黄酮(MHBFC)对血管紧张素Ⅱ(Ang Ⅱ)诱导的 H9c2 心肌细胞肥大的保护作用及可能机制.方法 将 H9c2 心肌细胞分为空白对照组、模型组、MHBFC 低、中、高剂量组,MHBFC 干预后,以浓度为 1 μmol/L 的 Ang Ⅱ诱导心肌细胞肥大模型.检测单位面积 H9c2 心肌细胞个数和平均表面积,TUNEL 法检测细胞凋亡,JC-1荧光探针法检测线粒体膜电位,酶联免疫吸附测定法(ELISA)检测心钠素(ANP)、脑钠尿肽(BNP)以及 β-肌球蛋白重链(β-MHC)含量,逆转录聚合酶链式反应法(RT-PCR)检测自噬标记因子 Beclin1 和 p62 的表达,蛋白免疫印迹法(Western blot)检测AMPK/mTOR 信号通路 p-AMPK/AMPK、p-mTOR/mTOR 和 p-ULK1/ULK1 表达水平.结果 MHBFC 能改善Ang Ⅱ诱导的心肌细胞数量下降以及心肌细胞肥大(P<0.05,P<0.01),TUNEL 法检测结果提示 MHBFC 能改善Ang Ⅱ诱导的细胞凋亡(P<0.05,P<0.01),JC-1荧光探针检测结果显示MHBFC 能改善Ang Ⅱ诱导的细胞线粒体膜电位下降并减少细胞凋亡(P<0.05,P<0.01),ELISA 法检测结果提示 MHBFC 可降低 Ang Ⅱ诱导的心肌细胞肥大标志物 ANP、BNP 以及 β-MHC 的水平(P<0.05,P<0.01),RT-PCR 检测结果显示 MHBFC 能上调 Beclin1 的表达(P<0.05,P<0.01)并降低 p62 的表达(P<0.05,P<0.01),Western blot 检测结果显示MHBFC 能提高 p-AMPK/AMPK、p-ULK1/ULK1 的蛋白表达(P<0.05,P<0.01)并下调 p-mTOR/mTOR 的蛋白表达(P<0.05,P<0.01).结论 MHBFC 能有效减轻 Ang Ⅱ诱导的 H9c2 心肌细胞肥大及损伤,其机制可能与激活 AMPK/mTOR 信号通路、促进线粒体自噬、减少心肌细胞肥大标志物的表达、抑制线粒体介导的细胞凋亡等有关.
Objective To investigate the protective effect of Yulangsan chalcone(MHBFC)on angiotensin II(Ang Ⅱ)-induced H9c2 cardiomyocytes hypertrophy and its possible mechanism in autophagy regulation through AMPK/mTOR signaling.Methods H9c2 cardiomyocytes were divided into blank control,model,and low-,medium-,and high-dose MHBFC groups.After intervention with MHBFC,Ang Ⅱ(1 μmol/L)was used to induce cardiomyocyte hypertrophy.The number and average surface area of H9c2 cardiomyocytes per unit area were detected,and cardiomyocyte apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling(TUNEL).Mitochondrial membrane potential was detected using a detection kit(JC-1 fluorescent probe method).The contents of atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),and β-myosin heavy chain(β-MHC)were detected by enzyme-linked immunosorbent assay(ELISA).The expression of autophagy markers Beclin1 and p62 was detected by reverse transcription-polymerase chain reaction(RT-PCR).The expression levels of p-AMPK/AMPK,p-mTOR/mTOR,and p-ULK1/ULK1 in the AMPK/mTOR signaling pathway were detected by Western blot.Results MHBFC improved the reduction of cardiomyocyte number and cardiomyocyte hypertrophy induced by Ang Ⅱ(P<0.05,P<0.01).TUNEL assay showed that MHBFC reduced Ang Ⅱ-induced apoptosis(P<0.05,P<0.01).MHBFC slowed the decrease of mitochondrial membrane potential and reduced AngⅡ-induced cell apoptosis(P<0.05,P<0.01).MHBFC reduced Ang Ⅱ-induced ANP,BNP,and β-MHC levels in cardiomyocytes(P<0.05,P<0.01).MHBFC increased Beclin1 expression(P<0.05,P<0.01)and reduced p62 expression(P<0.05,P<0.01).MHBFC increased p-AMPK/AMPK and p-ULK1/ULK1 protein expression(P<0.05,P<0.01)while reducing p-mTOR/mTOR expression(P<0.05,P<0.01).Conclusions MHBFC effectively alleviated Ang Ⅱ-induced H9c2 cardiomyocytes hypertrophy and injury;its mechanism may be related to activating AMPK/mTOR signaling,promoting mitophagy,reducing cardiomyocyte hypertrophy marker expression,and inhibiting mitochondria-mediated apoptosis.
农慧亮;黄湘;傅鹏;刘安韬;张婷;蒙泔竹;黄仁彬;梁杏梅;廖乃英
南宁市第一人民医院临床试验Ⅰ期研究室,南宁 530022广西卫生职业技术学院药学院,南宁 530023广西卫生职业技术学院药学院,南宁 530023广西卫生职业技术学院药学院,南宁 530023广西卫生职业技术学院药学院,南宁 530023广西卫生职业技术学院药学院,南宁 530023广西医科大学药学院,南宁 530021广西卫生职业技术学院药学院,南宁 530023广西卫生职业技术学院药学院,南宁 530023
医药卫生
玉郎伞黄酮心肌细胞肥大自噬H9c2心肌细胞
Yulangsan chalconecardiomyocyte hypertrophyautophagyH9c2 cardiomyocytes
《中国比较医学杂志》 2026 (18)
47-56,10
广西科技计划项目(桂科AB25069015,2023GXNSFBA026148)广西壮族自治区中医药管理局自筹经费科研课题(GXZYA20230245).
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