表皮生长因子对缺氧缺糖模型大鼠骨骼肌细胞氧化损伤的保护作用

doi:10.13481/j.1671-587x.20180219

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (02): 310-314.doi: 10.13481/j.1671-587x.20180219

表皮生长因子对缺氧缺糖模型大鼠骨骼肌细胞氧化损伤的保护作用

许雪梅¹, 黄笑夏², 金瓯¹, 张海邻³, 时洪雪⁴

1. 浙江省温州市中西医结合医院药剂科, 浙江温州 325000;
2. 浙江省温州市中医院药剂科, 浙江温州 325000;
3. 温州医科大学附属第二医院儿科, 浙江温州 325000;
4. 温州医科大学药学院临床药学系, 浙江温州 325000

收稿日期:2017-09-08 出版日期:2018-03-28 发布日期:2018-03-30
通讯作者: 许雪梅,主管药师(Tel:0577-85773087,E-mail:pharmacywz@163.com) E-mail:pharmacywz@163.com
作者简介:许雪梅(1980-),女,浙江省温州市人,主管药师,医学硕士,主要从事临床药学与药理机制方面的研究。
基金资助:
浙江省温州市科技局科技计划项目资助课题（Y20140649）；浙江省科技厅自然科学基金资助课题（LY17H010005，Y14H150023）

Protective effect of epithelial growth factor on oxidative damage of skeletal muscle cells in model rats of oxygen-glucose deprivation

XU Xuemei¹, HUANG Xiaoxia², JIN Ou¹, ZHANG Hailin³, SHI Hongxue⁴

1. Department of Pharmacy, Integrative Traditional Chinese Medicine and Western Medicine Hospital, Wenzhou City, Zhejiang Province, Wenzhou 325000, China;
2. Department of Pharmacy, Traditional Chinese Medicine Hospital, Wenzhou City, Zhejiang Province, Wenzhou 325000, China;
3. Department of Pediatrics Second Affiliated Hospital, Wenzhou Medical University, Wenzhou 325000, China;
4. Deparment of Clincal Pharmacy, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou 325000, China

Received:2017-09-08 Online:2018-03-28 Published:2018-03-30

摘要/Abstract

摘要： 目的：构建体外骨骼肌L6细胞缺氧缺糖（OGD）模型，在体外水平上探讨表皮生长因子（EGF）对压疮深部组织损伤（DTI）的保护机制。方法：将对数生长期大鼠骨骼肌L6成肌细胞分为5组，即正常对照组、OGD组、5 μg·L^-1 EGF+OGD组、10 μg·L^-1EGF+OGD组和20 μg·L^-1EGF+OGD组。MTT法检测各组细胞生存率，流式细胞术检测各组细胞凋亡率，DCFH-DA法检测各组L6成肌骨骼肌细胞中活性氧（ROS）水平，Rhodamine 123检测线粒体膜电势，Western blotting法检测各组骨骼肌细胞中Bcl-2和Bax蛋白表达。结果：与正常对照组比较，OGD组OGD24 h时骨骼肌细胞生存率明显降低（P<0.01），细胞凋亡率明显升高（P<0.01），ROS水平升高（P<0.01），线粒体膜电势下降（P<0.01），Bcl-2/Bax比值明显下降（P<0.01）。与OGD组比较，不同浓度EGF组细胞存活率明显升高，细胞凋亡率降低，其中10和20 μg·L^-1EGF组差异有统计学意义（P<0.05或P<0.01）；不同浓度EGF组骨骼肌细胞中ROS水平呈浓度依赖性降低，线粒体膜电势明显增加，10和20 μg·L^-1 EGF组差异有统计学意义（P<0.05或P<0.01）；Bcl-2/Bax比值明显下降，并具有浓度依赖性，其中10和20 μg·L^-1EGF组差异有统计学意义（P<0.05或P<0.01）。结论：EGF通过降低细胞内ROS水平保护线粒体功能，改善OGD诱导的大鼠骨骼肌细胞损伤，并具有浓度依赖性。

关键词: 骨骼肌L6成肌细胞, 活性氧簇, 缺氧缺糖, 表皮生长因子, 线粒体, 深部组织损伤

Abstract: Objective:To set up the rat skeletal muscle L6 cell models of oxygen-glucose deprivation (OGD) in vitro,and to investigate the protective effect of EGF in deep tissue injury(DTI) of pressure sores. Methods: The rat skeletal muscle cells in the logarithmic phase were divided into normal control group,OGD group,5 μg·L^-1 EGF+OGD group,10 μg·L^-1 EGF+ OGD group and 20 μg·L^-1 EGF+ OGD group.The survival rates of skeletal muscle cells in various groups were measured by MTT assay; the cell apoptotic rates in various groups were detected by flow cytometry; the reactive oxygen species (ROS) levels were detected by DCFH-DA; Rhodamine 123 was used to detect the mitochondrial membrane potential; the expressions of Bax and Bcl-2 proteins were determined by Western blotting method. Results: Compared with normal control group,the survival rates of skeletal muscle cells in OGD group after 24 h OGD was significantly decreased (P<0.05); the apoptotic rate was markedly increased (P<0.01); the ROS level was increased (P<0.01);the mitochondrial membrane potential was decreased (P<0.01); the ratio of Bcl-2/Bax was significantly decreased (P<0.01).Compared with OGD group, the survival rates of skeletal muscle cells in different concentrations of EGF groups were increased and the apoptotic rates were decreased,especially in 10 and 20 μg·L^-1 EGF groups (P<0.05 or P<0.01); the ROS levels in skeletal muscle cells in different concentrations of EGF groups were decreased and the mitochondrial membrane potential were increased,especially in 10 and 20 μg·L^-1 EGF groups (P<0.05 or P<0.01); the Bcl-2/Bax ratios were significantly decreased in a concentration-dependent manner,especially in 10 and 20 μg·L^-1 EGF groups(P<0.05 or P<0.01). Conclusion: EGF can improve the skeletal muscle cell injury induced by OGD in a concentration-dependent manner via decreasing the ROS levels and protecting the cell mitochondrial function.

Key words: deep tissue injury, reactive oxygen species, epithelial growth factor, skeletal muscle L6 cells, mitochondria, oxygen-glucose deprivation

中图分类号:

R966

许雪梅, 黄笑夏, 金瓯, 张海邻, 时洪雪. 表皮生长因子对缺氧缺糖模型大鼠骨骼肌细胞氧化损伤的保护作用[J]. 吉林大学学报(医学版), 2018, 44(02): 310-314.

XU Xuemei, HUANG Xiaoxia, JIN Ou, ZHANG Hailin, SHI Hongxue. Protective effect of epithelial growth factor on oxidative damage of skeletal muscle cells in model rats of oxygen-glucose deprivation[J]. Journal of Jilin University Medicine Edition, 2018, 44(02): 310-314.

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表皮生长因子对缺氧缺糖模型大鼠骨骼肌细胞氧化损伤的保护作用

Protective effect of epithelial growth factor on oxidative damage of skeletal muscle cells in model rats of oxygen-glucose deprivation

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