Novel RAS Inhibitors Poricoic Acid ZG and Poricoic Acid ZH Attenuate Renal Fibrosis via a Wnt/beta-Catenin Pathway and Targeted Phosphorylation of smad3 Signaling

Renal fibrosis is a common end point of the progression of chronic kidney disease (CKD). Suppressing the development and progression of renal fibrosis is essential in the treatment of kidney disease. Our previous study demonstrated that the ethyl acetate extract of the surface layer of Poria cocos exhibited beneficial antitubulointerstitial fibrosis. In this study, we isolated new diterpene (PZF) and triterpenes (PZG and PZH) and examined their antifibrotic effect. TGF-beta 1 upregulated the collagen I protein expression in HK-2 cells, and PZG and PZH treatment significantly inhibited the upregulated collagen I expression (TGF group 0.59 +/- 0.08 vs TGF+PZG group 0.36 +/- 0.08, P < 0.01; TGF+PZH group 0.39 +/- 0.12, P < 0.01). Triterpenes, PZG and PZH, exhibited a stronger inhibitory effect on renal fibrosis and podocyte injury than PZF. PZG and PZH further showed a stronger inhibitory effect on the activation of the renin angiotensin system (RAS) than PZF. Additionally, PZG and PZH markedly inhibited the activation of Wnt/beta-catenin signaling, which played an important role in fibrogenesis. Interestingly, PZG and PZH suppressed the TGF-beta/Smad pathway by selectively inhibiting the phosphorylation of Smad3 through blocking the interactions of SARA with TGF beta 1 and Smad3. The analysis of the structure-ctivity relationship demonstrated that their antifibrotic effects were closely associated with the first six-membered ring structure and the number of carboxyl groups in this type of compounds. Additionally, fifteen known triterpenes were identified. These novel tetracyclic triterpenoid compounds provided the potential lead compounds for the research and development of antifibrosis drug, and they possessed the potential to be utilized as RAS inhibitors.