Wang Meng-Yao, Gao Gai, Li Er-Wen, Zhang Xiao-Wei, Wang Hui, Xu Jiang-Yan, Zhang Zhen-Qiang, Wang Pan, Xie Zhi-Shen
Academy of Chinese Medical Sciences, Henan University of Chinese Medicine Zhengzhou 450046, China School of Pharmacy, Henan University of Chinese Medicine Zhengzhou 450046, China.
Academy of Chinese Medical Sciences, Henan University of Chinese Medicine Zhengzhou 450046, China.
Zhongguo Zhong Yao Za Zhi. 2022 Jan;47(2):453-460. doi: 10.19540/j.cnki.cjcmm.20211008.402.
The present study investigated the pharmaceutical effect and underlying mechanism of Zexie Decoction(ZXD) on nonalcoholic fatty liver disease(NAFLD) in vitro and in vivo via the LKB1/AMPK/PGC-1α pathway based on palmitic acid(PA)-induced lipid accumulation model and high-fat diet(HFD)-induced NAFLD model in mice. As revealed by the MTT assay, ZXD had no effect on HepG2 activity, but dose-dependently down-regulated alanine aminotransferase(ALT) and aspartate aminotransferase(AST) in the liver cell medium induced by PA, and decreased the plasma levels of ALT and AST, and total cholesterol(TC) and triglyceride(TG) levels in the liver. Nile red staining showed PA-induced intracellular lipid accumulation, significantly increased lipid accumulation of hepatocytes induced by PA, suggesting that the lipid accumulation model in vitro was properly induced. ZXD could effectively improve the lipid accumulation of hepatocytes induced by PA. Oil red O staining also demonstrated that ZXD improved the lipid accumulation in the liver of HFD mice. JC-1 staining for mitochondrial membrane potential indicated that ZXD effectively reversed the decrease in mitochondrial membrane potential caused by hepatocyte injury induced by PA, activated PGC-1α, and up-regulated the expression of its target genes, such as ACADS, CPT-1α, CPT-1β, UCP-1, ACSL-1, and NRF-1. In addition, as revealed by the Western blot and immunohistochemistry, ZXD up-regulated the protein expression levels of LKB1, p-AMPK, p-ACC, and PGC-1α in vivo and in vitro. In conclusion, ZXD can improve NAFLD and its mechanism may be related to the regulation of the LKB1/AMPK/PGC-1α pathway.
本研究基于棕榈酸（PA）诱导的脂质积累模型和高脂饮食（HFD）诱导的小鼠非酒精性脂肪性肝病（NAFLD）模型，通过LKB1/AMPK/PGC-1α通路，在体外和体内研究了泽泻汤（ZXD）对NAFLD的药理作用及潜在机制。MTT法检测结果显示，ZXD对HepG2细胞活性无影响，但能剂量依赖性下调PA诱导的肝细胞培养基中的丙氨酸氨基转移酶（ALT）和天冬氨酸氨基转移酶（AST），并降低血浆中ALT和AST水平以及肝脏中的总胆固醇（TC）和甘油三酯（TG）水平。尼罗红染色显示PA诱导细胞内脂质积累，显著增加PA诱导的肝细胞脂质积累，提示体外脂质积累模型诱导成功。ZXD可有效改善PA诱导的肝细胞脂质积累。油红O染色也表明ZXD改善了HFD小鼠肝脏中的脂质积累。线粒体膜电位的JC-1染色表明，ZXD有效逆转了PA诱导的肝细胞损伤所致的线粒体膜电位降低，激活了PGC-1α，并上调了其靶基因如ACADS、CPT-1α、CPT-1β、UCP-1、ACSL-1和NRF-1的表达。此外，蛋白质免疫印迹和免疫组织化学结果显示，ZXD在体内和体外均上调了LKB1、p-AMPK、p-ACC和PGC-1α的蛋白表达水平。综上所述，ZXD可改善NAFLD，其机制可能与调节LKB1/AMPK/PGC-1α通路有关。
