益气化瘀清热方对局灶节段性肾小球硬化小鼠足细胞线粒体自噬的调控机制研究
Study on the regulation mechanism of Yiqi Huayu Qingre Formula on mitochondrial autophagy of podocytes in focal segmental glomerular sclerosis mice
何改丽 1翟文生 1李冰 1任献青 1黄岩杰 1毕亮亮 1杨濛 1高旭光 1李广 1张秋月1
作者信息
- 1. 河南中医药大学第一附属医院,河南中医药大学儿科医学院,郑州 450000
- 折叠
摘要
目的:基于线粒体自噬探究局灶节段性肾小球硬化(FSGS)的发病机制及益气化瘀清热方的疗效机制.方法:运用两次尾静脉注射阿霉素法构建FSGS小鼠模型,小鼠随机分为空白组、模型组、益气化瘀清热方组、益气化瘀清热方高剂量组,除空白组外,其余各组分别于第1、5周尾静脉注射阿霉素,第2次尾静脉注射阿霉素后1周灌胃给药,共给药6周.透射电子显微镜观察小鼠肾脏病理、足细胞线粒体、线粒体自噬体等差异.qRT-PCR和Western Blot检测小鼠肾组织自噬相关蛋白ATG7、LC3B、Beclin1、p62 mRNA及蛋白的表达.结果:与空白组比较,模型组小鼠ATG7、LC3B和Beclin1蛋白表达显著升高(P＜0.05),LC3B、Beclin1 mRNA表达量显著升高(P＜0.05),p62蛋白及mRNA表达量显著降低(P＜0.05);与模型组比较,益气化瘀清热方组和益气化瘀清热方高剂量组小鼠肾组织自噬相关蛋白ATG7、LC3B、Beclin1蛋白及LC3B、Beclin1 mRNA表达显著升高(P＜0.05),p62蛋白及mRNA表达量显著降低(P＜0.05).结论:足细胞线粒体自噬是FSGS发生及进展的重要机制,益气化瘀清热方可通过增强线粒体自噬改善足细胞损伤.
Abstract
Objective:To explore the pathogenesis of focal segmental glomerular sclerosis(FSGS)and the therapeutic mechanism of Yiqi Huayu Qingre Formula based on mitochondrial autophagy.Methods:FSGS mouse model was constructed by twice doxorubicin injection into tail vein,and the mice were randomly divided into blank group,model group,Yiqi Huayu Qingre Formula group and Yiqi Huayu Qingre Formula high-dose group.Except the blank group,doxorubicin was injected into tail vein at the 1st and 5th week respectively.The therapeutic drugs were given by intragastric administration to the mice except the blank group after the second doxorubicin injection into tail vein for a total of 6 weeks.The differences of renal pathology,podocyte mitochondria and mitochondrial autophagosome were observed by transmission electron microscope.The expression of mRNA and protein of autophagy related proteins ATG7,LC3B,Beclin1 and p62 in renal tissue were detected by qRT-PCR and Western Blot.Results:Compared with blank group,the protein expressions of ATG7,LC3B and Beclin1 in model group were increased(P＜0.05),the mRNA expressions of LC3B and Beclin1 were significantly increased(P＜0.05),and the protein and mRNA expressions of p62 were significantly decreased(P＜0.05).Compared with the model group,the expressions of autophagy related proteins ATG7,LC3B and Beclin1 in renal tissue,mRNA expressions of LC3B,Beclin1 of Yiqi Huayu Qingre Formula group and Yiqi Huayu Qingre Formula high-dose group were significantly up-regulated(P＜0.05),while the expressions of p62 protein and mRNA were significantly down-regulated(P＜0.05).Conclusion:Podocyte mitochondrial autophagy is an important mechanism of the occurrence and progression of FSGS.Yiqi Huayu Qingre Formula can improve podocyte injury by enhancing mitochondrial autophagy.
关键词
局灶节段性肾小球硬化/肾组织/足细胞/线粒体自噬/益气化瘀清热方Key words
Focal segmental glomerular sclerosis(FSGS)/Kidney tissue/Podocyte/Mitochondrial autophagy/Yiqi Huayu Qingre Formula引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
万方数据