中国药理学通报2024，Vol.40Issue(1)：162-170,9.DOI:10.12360/CPB202308067
清肾颗粒对大鼠NRK-52E细胞转分化模型miR-23b和PINK1/Parkin通路的影响
Effect of Qingshen granules on miR-23b and PINK1/Parkin pathway in rat NRK-52E cell transdifferentiation model
摘要
Abstract
Aim To investigate the targeting mecha-nism of miR-23b on PINK1/Parkin pathway in transdif-ferentiation of NRK-52E cellsinduced by TGF-β1,and to elucidate the intervention mechanism of Qingshen granules drug-containing serum on NRK-52E cell transdifferentiation.Methods Ultra-high performance liquid chromatography(UPLC)fingerprinting method was used to analyze Qingshen granules.The NRK-52E transdifferentiation model induced by TGF-β1 was con-structed.The NRK-52E cells were divided into simula-ted no-load control group,miR-23b-5p simulated group,inhibitor no-load control group,and miR-23b-5p inhibitor group,after transfection with siRNA,and the effect of miR-23b-5p on PINK1 expression was ob-served.The NRK-52E cells were then divided into normal group,TGF-β1 group,Qingshen granule group,miR-23b-mimic group,miR-23b-mimic group,and miR-23b-mimic+Qingshen granule group.Western blot was used to detect the expression of Pink1,Parkin,LC3 Ⅱ,Beclin-1,P62 and α-SMA proteins,and RT-PCR was used to detect the expression of miR-23b-5p,Pink1,Parkin,Beclin-1 and α-SMA mRNA in NRK-52E cells.Dual-Luciferase Reporter gene experiment was used to detect the targeting relationship between miR-23b-5p and PINK1.Results UPLC fingerprint-ing method found 11 active components in Qingshen granules.After overexpression of miR-23b-5p,the ex-pression of PINk1 mRNA significantly increased(P＜0.05).And after silencing of miR-23b-5p expression,the expression of PINk1 mRNA also significantly de-creased(P＜0.05).Dual-Luciferase Reporter Assay showed that Rno-miR-23b-5p could significantly down-regulate the luciferase activity of Rno-PINK1-WT(P＜0.05),but could not down-regulate the luciferase ac-tivity of mutant Rno-PINK1-mut(P＞0.05).The ex-perimental results showed that the expressions of miR-23b-5p,Pink1,Parkin,Beclin-1,LC3 Ⅱ and LC3 Ⅱ/Ⅰratio in TGF-β1 group were significantly lower than those in normal group,but the expressions of P62 andα-SMA were significantly higher than those in normal group(P＜0.05).The expressions of miR-23b-5p,Pink1,Parkin,Beclin-1,LC3 Ⅱ and LC3 Ⅱ/Ⅰ ratio in Qingshen granule group and miR-23b-mimic group were significantly higher than those in TGF-β1 group,and the expressions of P62 and α-SMA were signifi-cantly lower than those in TGF-β1 group(P＜0.05).The performance of miR-23b-mimic+Qingshen granule group was better than that of miR-23b-mimic group(P＜0.05).Conclusions Qingshen granules can up-regulate the expression of miR-23b-5p in NRK-52E cellsand inhibit the transdifferentiation process of NRK-52E cells by enhancing the mitochondrial autophagy activity mediated by PINK1/Parkin pathway.关键词
miR-23b-5p/PINK1/Parkin信号通路/线粒体自噬/NRK-52E细胞/清肾颗粒/上皮细胞转分化Key words
MiR-23b-5p/PINK1/Parkin signaling pathway/mitophagy/NRK-52E cells/Qingshen gran-ules/epithelial-mesenchymal transition分类
医药卫生引用本文复制引用
金华,张叶青,呼琴,张磊,陈诺,韩燕全,王亿平..清肾颗粒对大鼠NRK-52E细胞转分化模型miR-23b和PINK1/Parkin通路的影响[J].中国药理学通报,2024,40(1):162-170,9.基金项目
国家自然科学基金资助项目(No 82274307,82004165) （No 82274307,82004165）
安徽省高等学校自然科学重点研究项目(No KJ2021A0546) （No KJ2021A0546）
