Huyan-I formula attenuates renal senescence and fibrosis by inhibiting STAT3/NF-κB/NLRP3-driven SASP

The Huyan-I formula (HY-I), a patented six‑herb traditional Chinese medicine used clinically at Jiangsu Provincial Hospital of TCM for early‑stage chronic kidney disease (CKD), has demonstrated therapeutic efficacy; however, its molecular mechanisms remain unclear. Given the central role of cellular senescence and fibrosis in CKD progression, particularly through the senescence‑associated secretory phenotype (SASP), this study investigated whether HY-I ameliorates renal senescence and fibrosis by modulating SASP‑related signaling. By UPLC‑Q‑TOF-MS analysis, 211 components were identified in HY‑I, and 29 prototype compounds plus 19 metabolites were detected in the serum of HY‑I‑treated mice. Network pharmacology and molecular docking predictions indicated interactions with the STAT3/NF‑κB/NLRP3/SASP axis. In naturally aged mice and in D‑galactose‑induced senescent human renal tubular (HK‑2) cells treated with HY-I or resveratrol (positive control), HY-I significantly reduced senescent cell burden and senescence markers, suppressed activation of STAT3, NF‑κB, and NLRP3, downregulated SASP expression, and improved tubular injury and renal fibrosis as assessed by Western blot, immunohistochemistry, multiplex immunofluorescence, and histopathological staining (HE, Masson, PAS). These findings demonstrate that HY-I alleviates renal aging and fibrosis by inhibiting the STAT3/NF‑κB/NLRP3 pathway and attenuating the SASP, thereby providing a mechanistic basis for its clinical application in CKD.