Leonurine Attenuates CCl4-Induced Hepatic Fibrosis in Mice via the Hippo-YAP Pathway

Pengru Wang,1,2,* Junjie Bai,1,2,* Mingxin Ye,2 Zhiwei Huang,1,2 Jiatong Chen,1,2 Lei Sun,1,2 Xiaolin Zhong,3 Yang Zheng,4 Tingting Ma,5 Wenguang Fu,1,2 Yichao Du1,6 1Metabolic Hepatobiliary and Pancreatic Diseases Key Laboratory of Luzhou City, Academician (Expert) Workstation of Sichuan Province, the Affiliated Hospital, Southwest Medical University, Luzhou, 646000, People’s Republic of China; 2Department of General Surgery (Hepatopancreatobiliary Surgery), the Affiliated Hospital, Southwest Medical University, Luzhou, 646000, People’s Republic of China; 3Department of Gastroenterology, The Affiliated Hospital of Southwest Medical University, Luzhou, People’s Republic of China; 4Drug Clinical Trial Institution, The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, People’s Republic of China; 5Clinical Medical Research Center, the Affiliated Hospital of Southwest Medical University, Luzhou, People’s Republic of China; 6Department of Gastroenterology, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, People’s Republic of China*These authors contributed equally to this workCorrespondence: Wenguang Fu, Department of General Surgery (Hepatopancreatobiliary Surgery), The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, People’s Republic of China, Tel +860830-3165200, Fax +8608302392753, Email [email protected] Yichao Du, Academician (Expert) Workstation of Sichuan Province, the Affiliated Hospital of Southwest Medical University, Luzhou, People’s Republic of China, Tel +860830-3165200, Fax +8608302392753, Email [email protected] and Purpose: The repair response to persistent damage in the liver manifests as liver fibrosis. Leonurine (Leo), an herbal remedy from traditional Chinese medicine, exhibits multiple biological activities such as antioxidant and anti-inflammatory effects. Nevertheless, research exploring Leo’s influence on liver fibrosis remains absent. This study aims to investigate the effect of Leo on liver fibrosis and clarify the mechanisms at play.Methods: We employed a mouse model of liver fibrosis induced by carbon tetrachloride (CCl4) and utilized LX-2 cells to investigate the mechanisms through which Leo exerts its effects.Results: Mice administered CCl4 displayed an elevated hepatic index, considerable necrosis as revealed by H&E staining, and significantly heightened serum markers of liver injury (ALT and AST). Conversely, mice treated with Leo showed a marked reduction in liver damage, coupled with diminished levels of inflammatory mediators and less infiltration of hepatic macrophages. Importantly, Leo demonstrated protective effects against CCl4-induced hepatocyte apoptosis in vivo and promoted apoptosis in hepatic stellate cells in vitro. Staining with Sirius red and Masson confirmed that Leo substantially reduced collagen deposition. Furthermore, Leo treatment led to decreased quantities of α-smooth muscle actin (α-SMA) and collagen type I (COL1A1) in both fibrotic tissues and LX-2 cells. Additionally, the administration of Leo was associated with a significant decrease in YAP protein levels, along with an increase in YAP phosphorylation at Ser 397 via the MST-LATS kinase signaling pathway, facilitating the degradation of YAP.Conclusion: The findings indicate that Leo may exert a protective effect against liver fibrosis through the inhibition of the Hippo-YAP signaling pathway, underscoring its potential as a therapeutic agent for treating hepatic fibrosis.Keywords: leonurine, liver fibrosis, hepatic stellate cells, apoptosis, cell cycle, yes-associated protein