南方医科大学学报2024，Vol.44Issue(8)：1431-1440,10.DOI:10.12122/j.issn.1673-4254.2024.08.01
清肾颗粒通过调控外泌体、miR-330-3p以及CREBBP表达抑制小鼠肾纤维化
Qingshen Granules alleviates renal fibrosis in mice by regulating exosomes,miR-330-3p,and CREBBP expression
摘要
Abstract
Objective To explore the effects of Qingshen Granules(QSG)on adenine-induced renal fibrosis in mice and in uric acid(UA)-stimulated NRK-49F cells and its mechanism for regulating exosomes,miR-330-3p and CREBBP.Methods A mouse model of adenine-induced renal fibrosis were treated daily with QSG at 8.0 g·kg-1·d-1 via gavage for 12 weeks.An adeno-associated virus vector was injected into the tail vein,and renal tissues of the mice were collected for analyzing exosomal marker proteins CD9,Hsp70,and TSG101 and expressions of Col-Ⅲ,α-SMA,FN,and E-cad using Western blotting and immunofluorescence and for observing pathological changes using HE and Masson staining.In the cell experiment,NRK-49F cells were stimulated with uric acid(400 μmol/L)followed by treatment with QSG-medicated serum from SD rats,and the changes in expressions of the exosomal markers and Col-Ⅲ,α-SMA,FN,and E-cad were analyzed.Dual luciferase reporter assay was employed to examine the targeting relationship between miR-330-3p and CREBBP,whose expressions were detected by RT-qPCR and Western blotting in treated NRK-49F cells.Results The mouse models of adenine-induced renal fibrosis showed significantly increased levels of CD9,Hsp70,and TSG101,which were decreased by treatment with QSG.The expressions of Col-Ⅲ,α-SMA,and FN increased and E-cad decreased in the mouse models but these changes were reversed by QSG treatment.QSG treatment obviously alleviated renal fibrosis in the mouse models.Intravenous injection of adeno-associated viral vector obviously inhibited miR-330-3p,increased CREBBP levels,and reduced fibrosis in the mouse models.Dual luciferase assay confirmed CREBBP as a target of miR-330-3p,which was consistent with the results of the cell experiments.Conclusion QSG inhibits renal fibrosis in mice by regulating the exosomes,reducing miR-330-3p levels,and increasing CREBBP expression.关键词
清肾颗粒/外泌体/miR-330-3p/肾纤维化Key words
Qingshen Granules/exosomes/miR-330-3p/renal fibrosis引用本文复制引用
戴荣,曹泽平,刘传娇,葛永,程梦,王伟丽,陈义珍,张磊,王亿平..清肾颗粒通过调控外泌体、miR-330-3p以及CREBBP表达抑制小鼠肾纤维化[J].南方医科大学学报,2024,44(8):1431-1440,10.基金项目
国家自然科学基金(82274307) （82274307）
合肥市科技局生命健康专项(GJ2022SM03) （GJ2022SM03）
2022临床医学研究转化专项(202104j07020014) （202104j07020014）
安徽省自然科学基金面上项目(2308085MH292,2208085MH269) （2308085MH292,2208085MH269）
安徽省高等学校科学研究重点项目(2023AH050749,2022AH050457,2022AH050465) （2023AH050749,2022AH050457,2022AH050465）
2023年度合肥综合性国家科学中心大健康研究院新安医学与中医药现代化研究所科研项目(2023CXMMTCM018)Supported by National Natural Science Foundation of China(82274307). （2023CXMMTCM018）
