丹七片调控mTOR-ULK1/TFEB自噬通路保护阿霉素诱导心肌损伤的作用机制  

Mechanism of Danqi Tablets protecting against doxorubicin-induced myocardial injury through regulating mTOR-ULK1/TFEB autophagy pathway

作　　者：王晓平 江艳艳 李伟利 孙乾斌 张倩 王勇 WANG Xiao-ping;JIANG Yan-yan;JI Weili;SUN Qian-bin;ZHANG Qian;WANG Yong(School of Traditional Chinese Mdicine,Bejig University of Chinese Medicine,Bejing 100209,China;School of Life Sciences,Bejing University of Chinese Medicine,Bejing 100292 China)

机构地区：[1]北京中医药大学中医学院,北京100029 [2]北京中医药大学生命科学学院,北京100029

出　　处：《中华中医药杂志》2021年第7期3946-3951,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81822049,No.81673712);霍英东教育基金项目(No.151044);北京科技新星项目(No.Z171100001117028);中华中医药学会青年托举工程(No.CACM-2017-QNRC2-C13)

摘　　要：目的:探讨丹七片对阿霉素诱导心肌损伤(DIMI)的保护作用及调控机制。方法:18只DIMI小鼠随机分成模型组、丹七片组和普伐他汀组,每组6只,另取6只C57BL/6小鼠作为正常对照组。丹七片组和普伐他汀组分别予1.5g/kg丹七片和40mg/kg普伐他汀混悬液灌胃,模型组和正常对照组予蒸馏水灌胃,连续干预4周。超声和HE检测心功能,电镜和GFP-mRFP-LC3腺病毒转染检测自噬通量,Western blot检测自噬相关蛋白的表达。结果:与模型组比较,丹七片组显著提高DIMI小鼠的射血分数(EF)和左室短轴缩短率(FS)(P<0.01,P<0.05);HE染色显示,丹七片有效改善心肌细胞排列紊乱和心肌纤维溶解的情况;电镜结果提示,丹七片组显著减少自噬溶酶体的累积。体外GFP-mRFP-LC3荧光结果表明,丹七片通过减少自噬溶酶体的累积和促进自噬体的形成恢复阿霉素阻断的自噬流;此外,丹七片组显著上调Beclin1和LAMP1(P<0.05),下调p-mTOR和p-ULK1(P<0.05),促进TFEB入核(P<0.01);联合mTOR激动剂MHY1485后,丹七片的调控作用均消失。结论:丹七片通过mTOR-ULK1/TFEB通路分阶段调控自噬保护DIMI。Objective: To investigate the protective effect and regulatory mechanism of Danqi Tablets on doxorubicininduced myocardial injury(DIMI). Methods: Eighteen DIMI mice were randomly divided into model group, traditional Chinese medicine group and positive drug group with 6 mice in each group, and 6 C57 BL/6 mice served as normal control group. The traditional Chinese medicine group and positive drug group were given 1.5 g/kg Danqi Tablets and 40 mg/kg Pravastatin suspension by gavage respectively, and the model group and blank control group were given distilled water by gavage for 4 weeks. Cardiac function was detected by Echocardiography and HE;autophagy flux was detected by electron microscopy and GFP-mRFP-LC3 adenovirus transfection;autophagy related protein expression was detected by Western blot. Results: Compared with the model group, Danqi Tablets significantly improved the ejection fraction(EF) and left ventricular short-axis shortening(FS) of DIMI mice(P<0.01, P<0.05). HE staining showed that Danqi Tablets effectively improved myocardium cell arrangement disorder and myocardial fiber lysis;electron microscopy results indicated that Danqi Tablets significantly reduced autolysosome accumulation;in vitro GFP-m RFP-LC3 fluorescence results showed that Danqi Tablets could reduce autolysosome accumulation and promote the formation of autophagosomes to restore the autophagic flux blocked by doxorubicin;In addition, Danqi Tablets significantly upregulated Beclin1 and LAMP1(P<0.05), down-regulated p-mTOR and p-ULK1(P<0.05), and promoted TFEB into the nucleus(P<0.01);combined with mTOR agonist MHY1485, the effect of Danqi Tablets disappeared. Conclusion: Danqi Tablets regulates autophagy and protects DIMI in stages through mTOR-ULK1/TFEB pathway.

关 键 词：阿霉素 心肌损伤 丹七片 自噬 mTOR-ULK1TFEB通路 中药复方 肿瘤心脏病 

分 类 号：R285.5[医药卫生—中药学]