Effect and Mechanisms of Yiqi Jianpi Xiaoyu Prescription on Kidney Fibrosis on the Basis of Metabolomics and Experimental Validation

The Yiqi Jianpi Xiaoyu prescription (YQJPXY) exhibits ameliorative effects on kidney fibrosis, but the underlying mechanism related to metabolism remains insufficient. We aimed to explore the antifibrotic effects of YQJPXY on kidney fibrosis and its underlying mechanism using serum metabolomics, biochemical analyses, and experimental analyses. We established a unilateral ureteral obstruction (UUO) kidney fibrosis mouse model, dividing mice into sham, model, YQJPXY-treated (7.28, 14.56, 29.12 g/kg/day), and Losartan groups. Renal function, histopathology, and fibrosis markers (FN, Col IV, and α-SMA) were analyzed. Serum metabolomic (UHPLC-Q-TOF/MS) identified key metabolites. TEM, qRT-PCR, WB, ELISA, and immunohistochemical validation confirmed the Adenosine mechanism in promoting autophagy through the A2BR/cAMP/AMPK pathway in the process of YQJPXY against kidney fibrosis. TGF-β-induced HK-2 cell fibrosis with autophagy inhibitor (CQ), A2BR inhibitor (PSB1115), and AMPK inhibitor (Dorsomorphin) explored the mechanism. YQJPXY improved kidney function and fibrosis. Metabolomics identified five important metabolites in kidney fibrosis: Adenosine monophosphate, Adenosine, Adenine, Inosine, and Hypoxanthine. Among these, Adenosine, which attenuated fibrosis via A2BR/cAMP/AMPK-mediated autophagy, was inhibited by CQ, PSB1115, and Dorsomorphin. YQJPXY's antifibrotic mechanism involves adenosine-activated A2BR/cAMP/AMPK autophagy pathways.