Original ArticlePuerarin Decreases Collagen Secretion in AngII-Induced Atrial Fibroblasts Through Inhibiting Autophagy Via the JNK–Akt–mTOR Signaling PathwayXu, Xudong MD; Jiang, Ruhong MD; Chen, Mengmeng MD; Dong, Mengmeng MS; Liu, Qiang MS; Cheng, Hui MS; Zhou, Kuangshi PhD; Chen, Laite MD; Li, Miaomiao MS; Jiang, Chenyang MD, PhDAuthor Information Department of Cardiology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, China. Reprints: Chenyang Jiang, MD, PhD, Department of Cardiology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, 3 East Qingchun Rd, Hangzhou, Zhejiang 310016, China (e-mail: [email protected]). Supported by grants from the National Natural Science Foundation of China (NO.81570296, awarded to C.J.), the Natural Science Foundation of Zhejiang Province (LQ17H020003, awarded to R.J.) and the Health Science and Technology Program of Hangzhou (2017A70, awarded to H.C.). The authors report no conflicts of interest. X. Xu and R. Jiang contributed equally to this work and should be considered co-first authors. Journal of Cardiovascular Pharmacology: June 2019 - Volume 73 - Issue 6 - p 373-382 doi: 10.1097/FJC.0000000000000672 Buy Metrics Abstract Puerarin is used to treat cardiovascular diseases due to its anti-inflammatory and antifibrotic effects. However, its mechanism of action in atrial fibroblasts is unknown. In this study, we investigated the autophagy pathway and molecular changes in angiotensin II (AngII)-stimulated atrial fibroblasts in response to puerarin treatment. Atrial fibroblasts were cultured and then subjected to stimulation with AngII and puerarin or other chemical drugs (3-MA, CQ, and SP600125). Quantitative real-time polymerase chain reaction and Western blot experiments were used to quantify the expression levels of mRNA and protein. mCherry-GFP-LC3 adenovirus was applied to reflect the autophagic flux. The results showed aggravating levels of autophagy and collagen deposit in the presence of AngII. Puerarin inhibited autophagy and decreased collagen secretion in a dose-dependent manner in atrial fibroblasts. Furthermore, phosphorylation of JNK was down-regulated in response to puerarin, whereas phosphorylation of Akt and mammalian target of rapamycin (mTOR) was upregulated. Interestingly, reduced autophagy and collagen secretion were observed when the JNK signaling pathway was blocked using SP600125. We also observed upregulation of Akt and mTOR phosphorylation in the presence of SP600125. These results suggest that puerarin exerts its antifibrotic effect in atrial fibroblasts partly through the inhibition of autophagy. Furthermore, the mechanism of action of puerarin in fibroblast autophagy seems to be mediated partly through JNK–Akt–mTOR signaling. Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.