益气化瘀清热方对阿霉素诱导损伤肾足细胞ATG7、LC3表达的影响
Effects of Yiqi Huayu Qingre Formula on the expression of ATG7 and LC3 in injured podocytes induced by adriamycin
李冰 1翟文生 1杨濛 2何改丽 2高旭光 2李广2
作者信息
- 1. 河南中医药大学儿科医学院,郑州 450000;河南中医药大学第一附属医院儿科医院,郑州 450000
- 2. 河南中医药大学第一附属医院儿科医院,郑州 450000
- 折叠
摘要
目的:观察益气化瘀清热方对阿霉素(ADR)诱导损伤肾足细胞自噬的影响.方法:制备益气化瘀清热方含药血清,采用ADR(0.2 μg/mL)诱导构建肾足细胞损伤模型,分为空白组、模型组(ADR组)、模型加含药血清组(ADR+Z组)、模型加空白血清组(ADR+K组).采用免疫荧光染色及Western Blot检测各组足细胞ATG7、LC3Ⅱ/Ⅰ表达,透射电镜观察足细胞内自噬体的形成以及足细胞线粒体的形态变化.结果:与空白组比较,ADR组与ADR+Z组LC3Ⅱ/Ⅰ、ATG7蛋白表达显著上调(P＜0.05,P＜0.01),与ADR组比较,ADR+Z组LC3Ⅱ/Ⅰ、ATG7蛋白表达显著增高(P＜0.01,P＜0.05).与空白组比较,ADR组有部分足细胞足突融合、脱落,肿胀变形的线粒体数量增多,细胞中自噬体数量增加.与ADR组比较,ADR+Z组足细胞足突数量增多,受损的线粒体数量明显减少,线粒体自噬体数量增多,溶酶体较多.结论:ADR损伤后,足细胞自噬水平反应性增强,线粒体损伤明显,益气化瘀清热方可上调ATG7、LC3Ⅱ/Ⅰ蛋白表达,增强ADR诱导足细胞线粒体自噬活动减轻足细胞损伤.
Abstract
Objective:To observe the effects of Yiqi Huayu Qingre Formula on the autophagy of injured podocytes induced by adriamycin(ADR).Methods:The serum containing Yiqi Huayu Qingre Formula was prepared.The injured podocytes model was induced by ADR(0.2 μg/mL)and divided into blank group,model group(ADR group),model plus drug-containing serum group(ADR+Z group)and model plus blank serum group(ADR+K group).The expressions of ATG7 and LC3Ⅱ/Ⅰ were detected by immunofluorescence staining and Western Blot.The formation of autophagosomes and morphological changes of mitochondria was observed by transmission electron microscopy.Results:The protein expression of LC3Ⅱ/Ⅰ and ATG7 increased in the ADR group and ADR+Z group compared with the blank group(P＜0.05,P＜0.01).Compared with the ADR group,the protein expression of LC3Ⅱ/Ⅰ and ATG7 significantly increased in the ADR+Z group(P＜0.01,P＜0.05).There were some fused and shed podocytes in the ADR group,and the number of swollen and deformed mitochondria and autophagosomes increased compared with the blank group.Compared with the ADR group,the number of foot processes,mitochondrial autophagosomes and lysosomes increased,and the damaged mitochondria decreased significantly in the ADR+Z group.Conclusion:After ADR injection,the level of autophagy in podocytes reactively increase and the damage to mitochondria is obvious.Yiqi Huayu Qingre Formula can up-regulate the expression of ATG7 and LC3Ⅱ/Ⅰ,and enhance the autophagy of mitochondria to reduce the injury of podocytes induced by ADR.
关键词
益气化瘀清热方/局灶节段性肾小球硬化/线粒体损伤/自噬/阿霉素/自噬相关蛋白7/微管相关蛋白1轻链3/足细胞Key words
Yiqi Huayu Qingre Formula/Focal segmental glomerulosclerosis(FSGS)/Mitochondria damage/Autophagy/Adriamycin(ADR)/ATG7/LC3/Podocytes引用本文复制引用
基金项目
国家自然科学基金项目(82104930)
国家自然科学基金项目(82274577)
国家自然科学基金项目(81873339)
河南省卫生健康委国家中医临床研究基地科研专项(2021JDZY103)
出版年
2024
国家科技期刊平台
NSTL
万方数据