Qingshen Granules alleviates renal fibrosis in mice by regulating exosomes,miR-330-3p,and CREBBP expression
10.12122/j.issn.1673-4254.2024.08.01
- VernacularTitle: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
- Author:
Rong DAI
1
;
Zeping CAO
;
Chuanjiao LIU
;
Yong GE
;
Meng CHENG
;
Weili WANG
;
Yizhen CHEN
;
Lei ZHANG
;
Yiping WANG
Author Information
1. 安徽中医药大学第一附属医院肾内科,安徽 合肥 230031
- Keywords:
Qingshen Granules;
exosomes;
miR-330-3p;
renal fibrosis
- From:
Journal of Southern Medical University
2024;44(8):1431-1440
- CountryChina
- Language:Chinese
-
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.
