槐耳浸膏对嘌呤霉素氨基核苷所致体外小鼠肾小球足细胞损伤的保护作用及其机制

doi:10.13481/j.1671-587x.20190508

吉林大学学报(医学版) ›› 2019, Vol. 45 ›› Issue (05): 1020-1024.doi: 10.13481/j.1671-587x.20190508

• 基础研究 • 上一篇

槐耳浸膏对嘌呤霉素氨基核苷所致体外小鼠肾小球足细胞损伤的保护作用及其机制

刘青菊¹, 高欣², 邱兵¹, 王燕³, 史凌娜³, 高霞⁴

1. 甘肃中医药大学研究生学院, 甘肃兰州 730000;
2. 甘肃省人民医院ICU科, 甘肃兰州 730000;
3. 宁夏医科大学研究生学院, 宁夏银川 750000;
4. 甘肃省人民医院儿科, 甘肃兰州 730000

收稿日期:2018-12-31 发布日期:2019-10-08
通讯作者: 高霞,副教授,主任医师,硕士研究生导师(Tel:0931-8281622,E-mail:gaoxia0219@163.com) E-mail:gaoxia0219@163.com
作者简介:刘青菊(1992-),女,四川省巴中市人,在读医学硕士,主要从事儿童肾脏疾病方面的研究。
基金资助:
国家自然科学基金地区项目资助课题（81360114，81660130）

Protective effect of extractum trametes robiniophila murr on puromycinaminonucleoside-induced podocyte injury in vitro of mice and its mechanism

LIU Qingju¹, GAO Xin², QIU Bing¹, WANG Yan³, SHI Lingna³, GAO Xia⁴

1. Graduate School, Gansu University of Chinese Medicine, Lanzhou 730000, China;
2. Department of ICU, People's Hospital of Gansu Province, Lanzhou 730000, China;
3. Graduate School, Ningxia Medical University, Yinchuan 750000, China;
4. Department of Pediatric, People's Hospital of Gansu Province, Lanzhou 730000, China

Received:2018-12-31 Published:2019-10-08

摘要/Abstract

摘要： 目的：探讨槐耳浸膏对嘌呤霉素氨基核苷（PAN）处理小鼠体外肾小球足细胞滤过率、活动力和细胞骨架重排的影响，阐明槐耳浸膏对足细胞损伤的保护作用及机制。方法：体外培养的足细胞随机分为对照组、模型组和实验组，其中模型组足细胞以50 mg·L^-1 PAN作用24 h，实验组足细胞经10 g·L^-1槐耳浸膏处理1 h后再换用含50 mg·L^-1 PAN的培养液作用24 h。采用两室弥散系统检测足细胞对异硫氰酸荧光素标记的白蛋白（FITC-BSA）的滤过率，细胞划痕实验检测足细胞划痕修复率，Transwell细胞迁移实验检测穿膜细胞数，激光共聚焦显微镜观察Invitrogen鬼笔环肽直接荧光标记足细胞纤维状肌动蛋白（F-actin）后细胞骨架的重排情况。结果：与对照组比较，模型组足细胞FITC-BSA滤过率明显升高（P<0.01）；与模型组比较，实验组足细胞FITC-BSA滤过率明显降低（P<0.01）。与对照组比较，模型组足细胞划痕修复率和穿膜细胞数均明显升高（P<0.05）；与模型组比较，实验组足细胞划痕修复率和穿膜细胞数均明显降低（P<0.05）。与对照组比较，模型组足细胞F-actin表达水平明显降低（P<0.01），F-actin重排率明显升高（P<0.01），足细胞骨架结构紊乱；与模型组比较，实验组足细胞F-actin表达水平明显升高（P<0.01），F-actin重排率明显降低（P<0.01），骨架重排情况明显缓解。结论：槐耳浸膏能够降低病理状态下的体外足细胞对牛血清白蛋白的滤过率，其机制可能与槐耳浸膏降低了足细胞活动力，进而改善体外足细胞骨架的重排有关。

关键词: 槐耳, 嘌呤霉素氨基核苷, 足细胞, 细胞骨架

Abstract: Objective:To investigate the effects of extractum trametes robiniophila murr on the filtration rate, motility and cytoskeleton rearrangement of the podocytes in vitro of the puromycin aminonucleoside(PAN)-treated mice, and to clarify the protective effect of extractum trametes robiniophila murr on the podocyte injury and its mechanism. Methods:The podocytes cultured in vitro were randomly divided into control group, model group and test group. The podocytess in model group were treated with 50 mg·L^-1 PAN for 24 h; the podocytess in test group were treated with 10 g·L^-1 extractum trametes robiniophila murr for 1 h and then treated with 50 mg·L^-1 PAN for 24 h. The filtration rate of podocytes to FITC-BSA was measured by two-compartment diffusion system; the scratch repair rate of podocytes was detected by cell scratch test, and the number of podocytes passing through the membrane was measured by Transwell cell migration test. Laser confocal microscope was used to observe the cytoskeleton rearrangement of podocyte cytoskeleton protein F-actin labeled with Invitrogen phalloidin directly. Results:Compared with control group, the FITC-BSA filtration rate of the podocytes in model group was increased significantly(P<0.01);compared with model group, the FITC-BSA filtration rate of the podocytes in test group was decreased significantly(P<0.01).Compared with control group, the scratch repair rate of podocytes and the number of transmembrane cells in model group were transmembrane(P<0.05);compared with model group, the scratch repair rate of podocytes and the number of migration migration cells in test group were decreased significantly(P<0.05). Compared with control group, the expression level of F-actin in the podocytes in model group was decreased significantly(P<0.01), the rearrangement rate of F-actin was increased signifiantly(P<0.01),and the structure of podocyte cytoskeleton was disordered;compared with model group, the expression level of F-actin in the podocytes in test group was increased significantly(P<0.01), the rearrangement rate of F-actin was decreased significantly(P<0.01),and the skeleton rearrangement was alleviated obviously. Conclusion:Extractum trametes robiniophila murr could reduce the filtration rate of podocytes to BSA in vitro under the pathological condition,and its possible mechanism is that extractum trametes robiniophila murr reduces the motility of podocytes and improve the rearrangement of podocyte cytoskeleton in vitro.

Key words: huaier, puromycin aminonucleoside, podocytes, cytoskeleton

中图分类号:

R692.6

刘青菊, 高欣, 邱兵, 王燕, 史凌娜, 高霞. 槐耳浸膏对嘌呤霉素氨基核苷所致体外小鼠肾小球足细胞损伤的保护作用及其机制[J]. 吉林大学学报(医学版), 2019, 45(05): 1020-1024.

LIU Qingju, GAO Xin, QIU Bing, WANG Yan, SHI Lingna, GAO Xia. Protective effect of extractum trametes robiniophila murr on puromycinaminonucleoside-induced podocyte injury in vitro of mice and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2019, 45(05): 1020-1024.

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槐耳浸膏对嘌呤霉素氨基核苷所致体外小鼠肾小球足细胞损伤的保护作用及其机制

Protective effect of extractum trametes robiniophila murr on puromycinaminonucleoside-induced podocyte injury in vitro of mice and its mechanism

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