清肾颗粒对5/6肾切除大鼠肾组织线粒体自噬及肾小管上皮-间质细胞转化的影响  

Effect of Qingshen Granule on Mitochondrial Autophagy in Renal Tissue and Renal Tubular EMT of 5/6 Nephrectomy Rats

作　　者：张叶青 金华[2] 张磊[2] 呼琴 王亿平[2] 代明扬 ZHANG Yeqing;JIN Hua;ZHANG Lei;HU Qin;WANG Yiping(Anhui University of Chinese Medicine,Anhui Hefei 230038,China;The First Affiliated Hospital of Anhui University of Chinese Medicine,Anhui Hefei 230031,China)

机构地区：[1]安徽中医药大学,安徽合肥230038 [2]安徽中医药大学第一附属医院,安徽合肥230031

出　　处：《中医药临床杂志》2024年第2期305-311,共7页Clinical Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82274307);国家自然科学基金青年基金项目(82004165);安徽省高等学校自然科学研究项目(重点项目)(KJ2021A0546)。

摘　　要：目的:探讨清肾颗粒对5/6肾切除大鼠肾组织线粒体自噬及肾小管上皮细胞转分化(Epithelial-mesenchymal,EMT)的影响。方法:雄性SD大鼠30只,随机分为假手术组、模型组、清肾颗粒组,每组各10只。除假手术组外,其余20只大鼠均制作5/6肾切除模型。清肾颗粒各组分别给予相应剂量清肾颗粒水溶液;假手术组、模型组以生理盐水灌胃。连续给药8周后,无菌采取腹主动脉血,代谢笼留取24h尿液,摘取左侧肾脏。检测血、尿肌酐浓度,并计算内生肌酐清除率;Western blot法检测肾组织中Pink1、Parkin、LC3-Ⅱ、α-SMA;免疫荧光法检测肾组织中LC3-Ⅱ和线粒体膜蛋白VDAC1共定位表达;HE和Masson染色光镜观察大鼠肾脏病理改变;透射电镜检测肾小管上皮细胞中线粒体的超微结构。结果:与假手术组比较,模型组大鼠的线粒体自噬相关蛋白Pink1、Parkin、LC3Ⅱ表达量均显著下降,肾小管EMT标志蛋白α-SMA显著升高(P<0.05)。与模型组比较,清肾颗粒组大鼠的Pink1、LC3Ⅱ蛋白表达量显著升高,α-SMA表达量显著降低(P<0.05),Parkin亦有所升高,但差异无统计学意义(P>0.05)。结论:清肾颗粒能够上调NRK-52E细胞内miR-23b-5p表达,并通过增强PINK1/Parkin通路介导的线粒体自噬活性,抑制NRK-52E细胞转分化进程。Objective:To investigate the effect of Qingshen Granules on mitochondrial autophagy and epithelial cell transdifferentiation(EMT)in renal tissue of 5/6 nephrectomized rats.Method:Thirty male SD rats were randomly divided into sham operation group,model group,and Qingshen Granule group,with 10 rats in each group.Except for the sham surgery group,all other 20 rats were made into a 5/6 nephrectomy model.Each group of Qingshen Granules was given corresponding doses of Qingshen Granule aqueous solution;The sham surgery group and model group were given physiological saline by gavage.After continuous administration for 8 weeks,sterile abdominal aortic blood was collected,and 24-hour urine was collected from the metabolic cage.The left kidney was removed.Detect blood and urine creatinine concentrations and calculate endogenous creatinine clearance rate;Western blot method for detecting Pink1,Parkin,LC3-Ⅱα-SMA;Immunofluorescence assay was used to detect the co localization expression of LC3-Ⅱand mitochondrial membrane protein VDAC1 in renal tissue;HE and Masson staining were used to observe the pathological changes of rat kidneys under light microscopy;Transmission electron microscopy was used to examine the ultrastructure of mitochondria in renal tubular epithelial cells.Result:Model group rat.Conclusion:Qingshen Granules can upregulate the expression of miR-23b-5p in NRK-52E cells and inhibit the process of NRK-52E cell transdifferentiation by enhancing the mitochondrial autophagy activity mediated by the PINK1/Parkin pathway.

关 键 词：清肾颗粒 5/6肾切除 线粒体自噬 PINK1/Parkin 肾小管上皮细胞转分化 

分 类 号：R692[医药卫生—泌尿科学]