Li Jie, Li Yang, Zhang Ying, Hu Dan, Gao Yonghong, Shang Hongcai, Xing Yanwei
Guang'anmen Hospital, Chinese Academy of Chinese Medical Sciences, Beijing 100053, China.
Key Laboratory of Chinese Internal Medicine of the Ministry of Education, Dongzhimen Hospital Affiliated to Beijing University of Chinese Medicine, Beijing 100700, China.
Oxid Med Cell Longev. 2017;2017:7042872. doi: 10.1155/2017/7042872. Epub 2017 Jun 20.
We investigated the role of cardiomyocyte autophagy and its regulatory mechanisms by WenxinKeli (WXKL) in cells subjected to hypertrophy.
H9C2 cardiomyocytes were divided into 8 groups. Cytoskeletal proteins as well as endogenously expressed autophagy marker proteins were studied by confocal imaging. Western blotting was used to assess the levels of light chain-3 (LC3) and mechanistic target of rapamycin (mTOR). The cell viability assay was used to detect the content of ATP. Flow cytometry was used to detect apoptotic cardiomyocytes.
(1) Compared with the control group, the length and width of cells in the Angiotensin II (AngII) group were significantly increased, while those in the 3-methyladenine (3-MA) and the WXKL groups were decreased. (2) Compared with AngII group, the expression of LC3 II/I protein in the 3-MA and WXKL groups was downregulated, while the expression of mTOR protein was upregulated. (3) Compared with the AngII group, the cardiomyocytes in the WXKL group showed increased ATP and decreased apoptosis rate and number of autophagosome.
We propose a novel role of WXKL as a likely inhibitor of cardiac hypertrophy by regulation of pathological autophagy.
我们研究了稳心颗粒（WXKL）在肥大细胞中对心肌细胞自噬的作用及其调控机制。
将H9C2心肌细胞分为8组。通过共聚焦成像研究细胞骨架蛋白以及内源性表达的自噬标记蛋白。采用蛋白质免疫印迹法评估轻链3（LC3）和雷帕霉素靶蛋白（mTOR）的水平。使用细胞活力测定法检测ATP含量。采用流式细胞术检测凋亡心肌细胞。
（1）与对照组相比，血管紧张素II（AngII）组细胞的长度和宽度显著增加，而3-甲基腺嘌呤（3-MA）组和WXKL组细胞的长度和宽度则减小。（2）与AngII组相比，3-MA组和WXKL组中LC3 II/I蛋白的表达下调，而mTOR蛋白的表达上调。（3）与AngII组相比，WXKL组的心肌细胞ATP增加，凋亡率和自噬体数量减少。
我们提出了WXKL作为一种可能通过调节病理性自噬来抑制心肌肥大的新作用。
