Jin-Gui-Shen-Qi Wan alleviates fibrosis in mouse diabetic nephropathy via MHC class II

作者全名："Liang, Dan; Liu, Lu; Qi, Yulin; Nan, Feng; Huang, Ju; Tang, Shiyun; Tang, Jianyuan; Chen, Nianzhi"

作者地址："[Liang, Dan; Tang, Jianyuan] Hosp Chengdu Univ Tradit Chinese Med, TCM Regulating Metab Dis Key Lab Sichuan Prov, Chengdu, Peoples R China; [Liu, Lu] Chengdu Univ Tradit Chinese Med, Coll Pharm, Chengdu, Peoples R China; [Qi, Yulin; Nan, Feng; Huang, Ju; Tang, Shiyun] Hosp Chengdu Univ Tradit Chinese Med, Chengdu, Peoples R China; [Chen, Nianzhi] Chongqing Med Univ, Coll Biomed Engn, State Key Lab Ultrasound Med & Engn, Chongqing, Peoples R China"

通信作者："Tang, JY (通讯作者)，Hosp Chengdu Univ Tradit Chinese Med, TCM Regulating Metab Dis Key Lab Sichuan Prov, Chengdu, Peoples R China.; Tang, SY (通讯作者)，Hosp Chengdu Univ Tradit Chinese Med, Chengdu, Peoples R China.; Chen, NZ (通讯作者)，Chongqing Med Univ, Coll Biomed Engn, State Key Lab Ultrasound Med & Engn, Chongqing, Peoples R China."

来源：JOURNAL OF ETHNOPHARMACOLOGY

ESI学科分类：PHARMACOLOGY & TOXICOLOGY

WOS号：WOS:001175873900001

JCR分区：Q1

影响因子：5.4

年份：2024

卷号：324

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：Jin-Gui Shen-Qi Wan; MHC class II; Diabetic nephropathy; Db/db mice; Kidney fibrosis

摘要："Ethnopharmacological relevance: Jin-Gui-Shen-Qi Wan (JGSQW) is a traditional Chinese medicine formula that has been traditionally used to alleviate urinary system ailments such as frequent urination and polyuria. Clinical studies have indicated that when combined with hypoglycaemic drugs, JGSQW exhibits a synergistic effect and can improve diabetic nephropathy (DN), yet its underlying mechanism and targets remain unclear. Aim of the study: This study aims to investigate the therapeutic efficacy of JGSQW and its underlying mechanisms using a DN db/db mouse model. Materials and methods: Ultrahigh-performance liquid chromatography coupled with mass spectrometry was utilized to analyse the primary active compounds, blood levels, and pharmacokinetics of JGSQW. Additionally, the therapeutic effects of JGSQW and metformin on blood glucose levels, lipid levels, renal function, and renal pathology in diabetic nephropathy mice were investigated using a db/db mouse model. Proteomic analysis was carried out to identify the primary target of JGSQW in treating DN. The mechanism of action was verified by western blotting, immunohistochemistry, and immunofluorescence. Then, molecular docking and molecular dynamics, transfection, drug affinity responsive target stability (DARTS) assay and cell thermal migration assay (CETSA) further validated the targeted binding effect. Results: JGSQW combined with metformin significantly improved the blood glucose levels, blood lipids, renal function, and renal pathology of DN mice. JGSQW mainly exerted its therapeutic effect on DN by targeting major histocompatibility complex class II (MHC class II) molecules. Immunohistochemistry results showed that JGSQW inhibited the expression of collagen I, fibronectin, and alpha smooth muscle actin (alpha-SMA) expression. Immunofluorescence and Western blot results showed that JGSQW inhibited the expression of H2-Ab1 and H2-Aa, which are MHC class II molecules, thereby suppressing CD4+ T-cell infiltration and improving diabetic kidney fibrosis. The binding ability of paeoniflorin to H2-Aa was predicted and verified by molecular, DARTS, and CETSA assays. Treatment with 80 mu M paeoniflorin effectively alleviated high glucose-induced injury in the MPC5 injury model. H2-Aa was overexpressed at this model concentration, and Western blotting further confirmed that paeoniflorin reduced glomerular podocyte fibrosis by regulating H2-Aa. Conclusions: JGSQW combined with metformin may have a synergistic effect to alleviates renal fibrosis in diabetic nephropathy by downregulating immune complex MHC class II molecules and attenuating the antigen presentation effect of MHC class II on CD4."

基金机构：National Natural Science Foundation of China [82004351]; Central Government Guides Local Science and Technology Development Projects of Sichuan Provincial Science and Technology Department [2021ZYD0107]; Xinglin Scholar Research Promotion Project of Chengdu University of TCM [QJRC2022008]

基金资助正文："<BOLD>Funding</BOLD> This work was supported by National Natural Science Foundation of China (Grant nos. 82004351) , and the Central Government Guides Local Science and Technology Development Projects of Sichuan Provincial Science and Technology Department (Grant no. 2021ZYD0107) , and the Xinglin Scholar Research Promotion Project of Chengdu University of TCM (Grant nos. QJRC2022008) ."