贝那鲁肽预处理对高糖诱导人视网膜色素上皮细胞损伤的影响
Effect of beinaglutide pretreatment on high glucose induced injury of human retinal pigment epithelial cells
矫梦瑶 1刘娟1
作者信息
- 1. 哈尔滨医科大学附属第一医院五官科手术室,黑龙江哈尔滨 150001
- 折叠
摘要
目的 探讨贝那鲁肽对高糖诱导人视网膜色素上皮(adult retinal pigment epithelial-19,ARPE-19)细胞损伤的影响及可能机制.方法 ARPE-19细胞分为4组,对照组细胞于正常葡萄糖(5 mmol/L)中培养,高糖组ARPE-19细胞于30 mmol/L葡萄糖中培养,贝那鲁肽低、高浓度组:用贝那鲁肽(50 nmol/L、100 nmol/L)培养.MTT法观察细胞增殖率,流式细胞计数检测凋亡率,Hoechst33258染色观察细胞凋亡,DCFH-DA染色检测细胞氧化应激水平,Western blot检测人磷酸化磷酸肌醇3激酶(phospho-rylation phosphotylinosital 3 kinase,p-PI3K)以及磷酸化苏氨酸蛋白激酶(phosphorylated serine-threonine protein kinase,p-AKT)的表达.结果 MTT结果显示,贝那鲁肽处理后ARPE-19细胞增殖率升高,与高糖组比较差异具有统计学意义(P＜0.05);高糖诱导ARPE-19细胞凋亡,贝那鲁肽抑制细胞凋亡,与高糖组比较差异有统计学意义(P＜0.05);DCFH-DA染色结果显示,贝那鲁肽逆转了高糖介导的ARPE-19细胞中活性氧(reactive oxygen species,ROS)的升高;Western blot印迹法结果显示,贝那鲁肽干预后p-PI3K及p-AKT蛋白表达上调,与高糖组相比差异均有统计学意义(P＜0.05).结论 贝那鲁肽通过调节氧化应激和细胞凋亡来减轻高糖引起的ARPE-19细胞损伤.
Abstract
Objective To investigate the effect of beinaglutide on high glucose induced damage to adult retinal pigment epithelial-19(ARPE-19)cells and its possible mechanism.Methods ARPE-19 cells were divided into four group.Cells in control group were cultured in normal glu-cose(5 mmol/L),cells in high glucose group were cultured in 30 mmol/L glucose,and cells in low and high concentration of beinaglutide were cultured with 50 nmol/L and 100 nmol/L benarulutide respectively.MTT method was used to observe cell proliferation rate,flow cytome-try was used to detect apoptosis rate,Hoechst33258 staining was used to observe cell apopto-sis,DCFH-DA staining was used to detect oxidative stress level,and Western blot was used to detect the expression of human phosphorylation phosphotylinosital 3 kinase(p-PI3K)and phos-phorylated AKT(p-AKT).Results The MTT results showed that the proliferation rate of ARPE-19 cells increased after treatment with beinaglutide,and the difference was statistically significant compared to the high glucose group(P＜0.05);High glucose induced apoptosis in ARPE-19 cells,while beinaglutide inhibited apoptosis,with a statistically significant difference compared with the high glucose group(P＜0.05);The results of DCFH-DA staining showed that beinaglutide reversed the increase of reactive oxygen species(ROS)in high glucose media-ted ARPE-19 cells.The results of Western blot showed that the expression of p-PI3K and p-AKT protein was upregulated after the intervention of beinaglutide,and the difference was sta-tistically significant compared with high glucose group(P＜0.05).Conclusion Beinaglutide alleviates high glucose induced damage to ARPE-19 cells by regulating oxidative stress and cell apoptosis.
关键词
贝那鲁肽/高糖/视网膜色素上皮细胞/氧化应激/凋亡Key words
benarutide/high glucose/retinal pigment epithelial cells/oxidative stress/apop-tosis引用本文复制引用
出版年
2024
NETL
NSTL
万方数据