Kangxianling Decoction Attenuates Renal Fibrosis by Inhibiting Ferroptosis

Kangxianling Decoction (KXL) is used clinically to attenuate renal fibrosis (RF). This study aimed to investigate the potential targets and molecular mechanisms of action of KXL in RF treatment. The chemical profile of KXL was initially characterized using UPLC-HRMS. Based on the identified core bioactive components, a network pharmacology approach was employed to predict potential targets and signaling pathways. A rat model of 5/6 nephrectomy (5/6 N) was established to assess the anti-RF effects of KXL. Hematoxylin and eosin (HE) and Masson's trichrome staining were performed to assess tissue morphology. A cellular ferroptosis model was established in HK-2 cells to validate the predicted signaling pathways and targets of KXL by immunofluorescence and western blotting. UPLC-HRMS identified eight major bioactive components in KXL, and subsequent network pharmacology analysis suggested that ferroptosis and the p53 signaling pathway might be implicated in the mechanism of RF. vidence of ferroptosis was confirmed by abnormal levels of ferroptosis-related markers (reactive oxygen species superoxide dismutase, glutathione, and morphological changes in mitochondria). The anti-RF effect of KXL is associated with the inhibition of ferroptosis by downregulating the expression level of P53 and enhancing antioxidant capacity. KXL reversed the downregulation of GPX4 caused by ferroptosis. Our findings indicate that the protective effect of KXL against RF is associated with the inhibition of ferroptosis via regulating p53/GPX4 pathway.