miR-125a-5p通过黏着斑通路对铝染毒GC2-spd细胞凋亡的影响  

Effects of miR-125a-5p on the apoptosis of aluminum-exposed GC2-spd cells through focal adhesion pathway

作　　者：黄艳新[1,2] 陈文成 彭慧鑫[2] 韦光吉 庞艳芳 元辉雄 HUANG Yanxin;CHEN Wencheng;PENG Huixin;WEI Guangji;PANG Yanfang;YUAN Huixiong(Department of Medical Laboratory,Affiliated Hospital of Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China;Graduate School,Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China)

机构地区：[1]右江民族医学院附属医院医学检验科,广西百色533000 [2]右江民族医学院研究生学院,广西百色533000

出　　处：《右江医学》2023年第5期397-402,共6页Chinese Youjiang Medical Journal

基　　金：广西自然科学基金(2020GXNSFAA297257);广西高校中青年教师科研基础能力提升项目(2020KY13017);广西壮族自治区中医药管理局自筹经费科研课题(GZZC2020248);广西壮族自治区卫生健康委员会自筹经费科研课题(Z20201416)。

摘　　要：目的 探讨miR-125a-5p在铝染毒小鼠精母细胞系(GC2-spd)模型中对黏着斑信号通路及GC2-spd细胞凋亡的影响。方法 建立铝染毒GC2-spd细胞模型,分为对照组(CG),铝染毒组(ALG),铝染毒组+miR-125a-5p inhibitor阴性对照组(ALG+inhibitor NC)及铝染毒组+miR-125a-5p inhibitor组(ALG+inhibitor);利用RT-qPCR检测ALG与CG细胞中miR-125a-5p的表达情况;利用Western blot检测CG组、ALG组、ALG+inhibitor NC组及ALG+inhibitor组Itga7、Pak4、Akt1、Bax及Caspase-9的表达情况;利用CCK8实验及流式细胞术实验检测各组GC2-spd细胞的活性和凋亡率。结果 RT-qPCR结果显示,与对照组比较,ALG组细胞中miR-125a-5p的表达显著升高;Western blot结果显示,抑制miR-125a-5p的表达后,可显著升高铝染毒GC2-spd细胞中Itga7、Pak4和Akt1的表达,降低Bax及Caspase-9的表达;CCK8和流式细胞术实验结果显示,抑制miR-125a-5p的表达后,可显著提高GC2-spd细胞的活性,降低细胞的凋亡率。结论 miR-125a-5p inhibitor可通过提高Itga7、Pak4和Akt1的表达,促进黏着斑信号通路激活,抑制细胞凋亡,提高细胞的活性,从而抑制铝染毒GC2-spd细胞的凋亡作用。Objective To investigate the effect of miR-125a-5p on focal adhesion signaling pathway and apoptosis of GC2-spd cells in aluminum-exposed mouse spermatocyte cell line(GC2-spd)model.Methods Aluminum-exposed GC2-spd cell model was established,and cells were divided into control group(CG),aluminum exposure group(ALG),ALG+miR-125a-5p inhibitor negative control(ALG+inhibitor NC)group,and ALG+miR-125a-5p inhibitor(ALG+inhibitor)group.The expressions of miR-125a-5p in the ALG and the CG were detected by RT-qPCR;the expressions of Itga7,Pak4,Akt1,Bax and Caspase-9 in the CG,the ALG,the ALG+inhibitor NC group and the ALG+inhibitor group were detected by Western blot;and the activity and apoptosis rate of GC2-spd cells in each group were detected by CCK-8 assay and flow cytometry.Results RT-qPCR results showed that the expression of miR-125a-5p in ALG cells was significantly increased while compared with that of the normal control group;Western blot results showed that inhibiting the expression of miR-125a-5p could significantly increase the expressions of Itga7,Pak4 and Akt1,and decrease the expressions of Bax and Caspase-9 in aluminum-exposed GC2-spd cells;the results of CCK8 and flow cytometry experiments showed that inhibiting the expression of miR-125a-5p could significantly increase the activity of GC2-spd cells and reduce the apoptosis rate of cells.Conclusion miR-125a-5p inhibitor can promote the activation of focal adhesion pathway,inhibit cell apoptosis,and improve cell activity by increasing Itga7,Pak4 and Akt1,thereby inhibiting the apoptosis of GC2-spd cells exposed to aluminum.

关 键 词：miR-125a-5p 黏着斑通路 铝染毒 小鼠精母细胞系细胞 

分 类 号：R114[医药卫生—卫生毒理学]