Wenyang Xiaozheng Decoction attenuates renal fibrosis and suppresses pyroptosis-associated signaling: insights from integrated transcriptomics and serum pharmacochemistry.

ETHNOPHARMACOLOGICAL RELEVANCE Wenyang Xiaozheng Decoction (WXD) is traditionally prescribed for chronic kidney disease based on the principle of warming yang and resolving blood stasis. However, whether its anti-fibrotic effects involve inflammation-associated pyroptosis remains unclear.

Aim

OF THE STUDY To evaluate the effects of WXD on renal fibrosis and associated changes in NF-κB- and pyroptosis-related signaling.

Materials And Methods

Thirty-six male C57BL/6 mice were allocated to six groups and subjected to sham surgery or unilateral ureteral obstruction (UUO) with or without valsartan or graded doses of WXD. The effects of WXD were further evaluated in TGF-β1/TNF-α-stimulated HK-2 cells. Kidney transcriptomics was integrated with serum pharmacochemistry-based network pharmacology restricted to blood-absorbed constituents. Pharmacological perturbations using necrosulfonamide (NSA) and lipopolysaccharide (LPS) were performed in HK-2 cells.

Results

High-dose WXD reduced serum BUN, renal fibrotic area, and GSDMD-N expression by 24.7%, 27.6%, and 29.1%, respectively, compared with UUO mice (all P < 0.05). Transcriptomic analysis showed that WXD reversed UUO-induced activation of NF-κB, TNF, and NOD-like receptor signaling. Integration with serum pharmacochemistry identified 36 blood-absorbed constituents and yielded inflammation-related target and pathway profiles consistent with the transcriptomic findings. WXD reduced the abundance of phosphorylated p65, cleaved caspase-1, and GSDMD-N in vivo and in vitro. NSA partially reproduced, whereas LPS partially counteracted, the protective effects of WXD.

Conclusions

WXD attenuates renal fibrosis in association with reduced NF-κB- and pyroptosis-associated signaling. Integrating transcriptomics with network pharmacology based on blood-absorbed constituents provides a biologically relevant strategy for investigating multi-component traditional Chinese medicine formulas.