中国病理生理杂志2024，Vol.40Issue(8)：1479-1487,9.DOI:10.3969/j.issn.1000-4718.2024.08.015
清肾颗粒通过miR-4516/SIAH3/PINK1调节线粒体自噬而减轻大鼠肾纤维化
Qingshen granules regulate mitophagy through miR-4516/SIAH3/PINK1 to attenuate renal fibrosis in rats
摘要
Abstract
AIM:To explore the mechanism by which Qingshen granules(QSG)intervene in the microRNA-4516(miR-4516)targeted regulation of the SIAH3/PINK1 axis,enhancing mitophagy and inhibiting renal fibrosis.METHODS:Male SD rats were randomly divided into three groups:control,model,and QSG groups.The QSG aqueous solution was administered via gavage once daily,4 mL each time,for 8 consecutive weeks.Blood creatinine levels were measured in each group.Hematoxylin-eosin(HE)and Masson staining were utilized to assess the degree of renal patholog-ical damage.Western blot analysis was performed to determine the expression levels of β-actin,PINK1,Parkin,SIAH3,VDAC1,Mfn1,Mfn2,OPA1,LC3B,and P62 proteins in renal tissue.RT-qPCR was used to detect the mRNA expres-sion level of SIAH3 in rat kidney tissue,and transmission electron microscopy was employed to observe mitochondrial dam-age in renal tissue.QSG-containing serum and transforming growth factor-β1(TGF-β1)were used to induce an HK-2 cell fibrosis model.The cells were divided into the following groups:normal cell(NC)group,model cell(MC)group,MC+miR-4516 mimics group,MC+miR-4516 NC+QSG group,MC+miR-4516 mimics+QSG group,and MC+QSG group.Cell activity in each group was detected using the CCK-8 method,and Western blot analysis was performed to determine E-cad-herin and α-SMA protein expression levels.The regulation of SIAH3 by miR-4516 was verified using a dual luciferase re-porter assay.RT-qPCR was used to detect the mRNA expression of miR-4516,SIAH3 mRNA,and PINK1.RESULTS:The results indicated that QSG intervention reduced fibrosis in rat renal tissue and HK-2 cells,decreased SIAH3 mRNA expression,increased PINK1 expression,and activated mitophagy in renal tissue.In vitro results confirmed that QSG can elevate miR-4516 expression,inhibit SIAH3 mRNA expression,promote PINK1 expression in HK-2 cells,and reduce the expression of the fibrosis marker protein α-SMA.CONCLUSION:In summary,this study preliminarily clarified the mechanism by which QSG intervention targets miR-4516 to regulate the SIAH3/PINK1 axis,thereby enhancing mitophagy and inhibiting renal fibrosis.关键词
清肾颗粒/微小RNA-4516/SIAH3/PINK1轴/线粒体自噬/肾纤维化Key words
Qingshen granules/microRNA-4516/SIAH3/PINK1 axis/mitophagy/renal fibrosis分类
医药卫生引用本文复制引用
任克军,金华,王东,茅燕萍,戴荣,程梦,王亿平,张磊..清肾颗粒通过miR-4516/SIAH3/PINK1调节线粒体自噬而减轻大鼠肾纤维化[J].中国病理生理杂志,2024,40(8):1479-1487,9.基金项目
安徽省高等学校科学研究重点项目(No.KJ2021A0545 （）
No.2023AH050749) （）
安徽省自然科学基金资助项目(No.2208085MH269) （No.2208085MH269）
2022临床医学研究转化专项(No.202104j07020014) （No.202104j07020014）
