Yi-shen-hua-shi granules modulate immune and inflammatory damage via the ALG3/PPARγ/NF-κB pathway in the treatment of immunoglobulin a nephropathy.

Controversy persists regarding the treatment of immunoglobulin A nephropathy (IgAN), thereby highlighting the demand for safer more effective therapeutic drugs. Although supplementary treatment using Yi-Shen-Hua-Shi (YSHS) granules has distinct advantages with respect to improving renal function in IgAN, a lack of clarity regarding the underlying mechanisms limits their clinical application. In this study, we aimed to elucidate the therapeutic mechanisms underlying the efficacy of YSHS granules in the treatment of IgAN. A rat model of IgAN was established based on lipopolysaccharide, carbon tetrachloride, and bovine serum albumin induction. In order to evaluate the effects of YSHS granules, we performed a range of techniques, including immunofluorescence assays, hematoxylin and eosin staining, and flow cytometry, to assess inflammation, immunity, and other relevant factors. Direct data-independent acquisition–mass spectrometry (DIA-MS) analysis and parallel reaction monitoring (PRM) were used for functional characterization and quantitative validation of differentially expressed proteins (DEPs), and Western blot analysis is used to identify downstream proteins associated with DEPs. Compared with the model group, the levels of proteinuria, urine red blood cells, serum creatinine, blood urea nitrogen, low-density lipoprotein-cholesterol, triglycerides, and pathological kidney damage were reduced in the YSHS group. A high dose of YSHS granules was found to raise the levels of CD8 T cells and reduce the CD4/CD8 ratio in the peripheral serum. To examine the mechanisms underlying the therapeutic effects YSHS granules, we performed direct DIA-MS analysis to identify proteins that were differentially expressed among the model, YSHS, and control groups. A total of 29 proteins were identified as being commonly expressed in all three groups. Further KEGG and protein–protein interaction (PPI) network analysis revealed that YSHS granules can contribute to the regulation of N -glycosylation-associated proteins, such as ALG3 and STT3A, in rats with IgAN. Detected changes in the expression of ALG3 and STT3A were consistent with the PRM results. We also established that the administration of YSHS granules can contribute to regulation of the ALG3-associated PPAR-γ/NF-κB signaling pathway. Our findings in this study provide evidence to indicate the efficacy of YSHS granules in the treatment of IgAN, the putative underlying mechanisms of which involve the modulation of N-glycosylation, mediated via the PPAR-γ/NF-κB pathway. A detailed flow chart of the study design. The effects of YSHS granules on immune disorders, inflammatory injury, and metabolic disorders in IgAN rats were assessed using a rat model in which IgAN was induced by lipopolysaccharide (LPS), carbon tetrachloride (CCl4), and bovine serum albumin (BSA). To investigate the mechanisms underlying the therapeutic effects of YSHS granules, we performed direct data-independent acquisition mass spectrometry (DIA-MS) analysis. Further Kyoto Encyclopedia of Genes and Genomes and protein–protein interaction network analysis indicated that YSHS granules regulate N-glycosylation-associated proteins, including ALG3 and STT3A, in rats with IgAN. On the basis of Western blot analysis, we demonstrated that YSHS granules regulate the ALG3-associated PPAR-γ/NF-κB signaling pathway. [Display omitted] • YSHS granule improves IgAN renal function but mechanism unknown. • Proteomic data reveals 29 targets and N-glycosylation in YSHS against IgAN. • ALG3, a key protein，regulates immune and inflammation related proteins. • YSHS granule regulates ALG3/PPAR-γ/NF-κB in the treatment of IgAN. [ABSTRACT FROM AUTHOR]