Integrated proteomics and bioinformatics reveal the pharmacological mechanism of Shenqi Yanshen granule in chronic kidney disease-mineral and bone disorder (CKD-MBD).

ETHNOPHARMACOLOGICAL RELEVANCE: Shenqi Yanshen Granule (SYG), a traditional Chinese medicine formulation, is clinically used for chronic kidney disease-mineral bone and disorder (CKD-MBD) by tonifying the kidney, activating blood, eliminating toxins, and dispelling turbidity. However, its mechanism remains unclear.

AIM OF THE STUDY: This study aimed to elucidate the therapeutic mechanism of SYG in CKD-MBD using an integrated approach combining proteomics and bioinformatics.

MATERIALS AND

METHODS

Forty-four CKD-MBD patients were divided into a treatment group (TG, n = 23) receiving standard therapy plus SYG (30g, twice daily), or a control group (CG, n = 21) receiving standard therapy alone. A normal group (NG, n = 20) served as healthy controls. After 8 weeks, renal function, mineral-bone metabolism parameters, and bone mineral density (BMD) were evaluated. Plasma proteomics was used to identify potential targets, followed by network pharmacology, molecular docking, and dynamics simulations to predict bioactive compounds. Finally, RANKL-stimulated RAW264.7 cells were conducted to validate the therapeutic targets and bioactive compounds.

RESULTS

After 8 weeks of treatment in a small cohort, preliminary improvements were observed in renal function (Scr, BUN), mineral-bone metabolism markers (Ca, iPTH, 25(OH)VD, b-ALP, β-CTX), and BMD in CKD-MBD patients. Plasma proteomics indicated that SYG downregulated PAK2 and CRKL within the MAPK pathway. Network pharmacology identified icariin as a constituent capable of binding both targets, which was supported by molecular docking and dynamics simulations showing stable complex formation under static and dynamic conditions. In vitro, both SYG and icariin significantly suppressed PAK2 and CRKL expression at mRNA and protein levels during RANKL-induced osteoclast differentiation in RAW264.7 cells.

CONCLUSION

This study suggests that PAK2 and CRKL are potential therapeutic targets of SYG for ameliorating CKD-MBD, and icariin may be an important active ingredient. These findings provide a foundational mechanistic hypothesis that warrants further validation in larger-scale clinical trials.