归脾汤通过Bmal1调控高糖状态下H9c2心肌细胞自噬活性的研究  

GuiPi Decoction(归脾汤)Regulates Autophagy of H9c2 Cardiomyocytes Exposed to High Glucose via Bmal1

作　　者：梁一超 陈诗成 李晓文 陆欣雨 黄颖怡 朱佳佳[2] 李俊[2,3] Liang Yichao;Chen Shicheng;Li Xiaowen;Lu Xiny;Huang Yingyi;Zhu Jiajia;Li Jun(TCM-Integrated Hospital,Southern Medical University,Guangzhou 510000;Nanfang Hospital,Southern Medical University,Guangzhou 510515;School of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515;Guangzhou First People's Hospital,Guangzhou 510180)

机构地区：[1]南方医科大学中西医结合医院,广州510000 [2]南方医科大学南方医院,广州510515 [3]南方医科大学中医药学院,广州510515 [4]广州市第一人民医院,广州510180

出　　处：《中药药理与临床》2022年第6期42-47,共6页Pharmacology and Clinics of Chinese Materia Medica

基　　金：广东省科技计划项目(编号:2016A020226026);广东省中医药局面上项目(编号:20161167)。

摘　　要：目的:探讨归脾汤在高糖状态对H9c2心肌细胞自噬活性的影响及脑和肌肉芳香烃受体核转运样蛋白1基因(Bmal1)在此过程中的作用。方法:将H9c2心肌细胞随机分为空白对照组、模型对照组、归脾汤2.5、7.5、15μg/mL组,共培养48 h。以Hochest33342/PI染色计算凋亡率;Western blot检测心肌细胞天冬氨酸蛋白水解酶3(CC3)、Bmal1及自噬相关蛋白(ATG)如ATG3、ATG5、ATG7及微管相关蛋白1轻链3β(LC3B)等的表达。抑制Bmal1表达后,观察H9c2心肌细胞凋亡率及自噬相关蛋白表达的变化。结果:与空白对照组比,模型对照组CC3蛋白表达上调,细胞凋亡率显著升高(P<0.01),同时Bmal1及自噬相关蛋白表达显著下调(P<0.01);与模型对照组比,各给药组CC3蛋白表达下调,归脾汤7.5、15μg/mL组细胞凋亡率显著下降(P<0.01),而自噬相关蛋白表达显著上调(P<0.01);抑制心肌细胞Bmal1的表达后,各给药组CC3蛋白表达上调,细胞凋亡率显著上升(P<0.01),自噬相关蛋白表达受到显著抑制(P<0.01)。结论:归脾汤可增强在高糖状态下心肌细胞的自噬活性,减少心肌细胞凋亡,其机制可能与上调Bmal1蛋白有关。Objective:To investigate the effect of GuiPi Decoction(归脾汤,GPD)on the autophagy of the cardiomyocytes exposed to high glucose(HG)and the role of brain and muscle Arnt-like protein-1(Bmal1)in this process.Methods:H9 c2 cardiomyocytes were randomely assigned into a blank control group,a model group,and low-,medium-,and high-dose(2.5,7.5,and 15μg/mL,respectively)GPD groups and cultured for 48 h.Hochest33342/PI staining was adopted to calculate the apoptosis rate.Western blotting was employed to determine the expression levels of cardiomyocyte caspase-3(CC3),BMAL1,autophagy-related proteins(ATG3,ATG5,and ATG7),and microtubule-associated protein 1 light chain 3-beta(LC3 B).After the expression of Bmal1 was inhibited,the apoptosis rate of H9 c2 cardiomyocytes and the expression levels of autophagy-related genes were measured.Results:Compared with the blank control group,HG treatment increased the apoptosis rate,up-regulated the expression of CC3,and down-regulated the expression of Bmal1 and autophagy-related proteins(P<0.01).Compared with the model group,GPD down-regulated the expression of CC3.GPD at 7.5μg/mL and 15μg/mL decreased the apoptosis rate and up-regulated the expression of autpphagy-related proteins in cardiomyocytes(P<0.01).The inhibition on the expression of Bmal1 increased the apoptosis rate,up-regulated the expression of CC3,and down-regulated the expression of autophagy-related proteins in the GPD groups(P<0.01).Conclusion:GPD might protect cardiomyocytes from the injury induced by HG via Bmal1-mediated activation of apoptosis and autophagy.

关 键 词：归脾汤 糖尿病心肌病 自噬 生物钟基因-脑和肌肉芳香烃受体核转运样蛋白1 

分 类 号：R285.5[医药卫生—中药学]