黄艳新 陈文成 彭慧鑫 韦光吉 庞艳芳 元辉雄

【摘要】 目的 探討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。

【关键词】 miR-125a-5p；黏着斑通路；铝染毒；小鼠精母细胞系细胞

中图分类号：R114 文献标志码：A DOI：10.3969/j.issn.1003-1383.2023.05.003

【Abstract】 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.

【Key words】 miR-125a-5p; focal adhesion pathway; aluminum exposure; GC2-spd cells

据世界卫生组织统计，全世界正常生育的夫妇中，就有10%～15%的夫妇面临不孕不育问题，其中大约有50%的病例是由男性因素引起的［1］。导致男性不育的因素有很多，致病机制尚未完全明确，由于精子发生的过程受到很多遗传和外界环境因素的影响，精液质量低下可能与先天性发育异常、免疫、感染和内分泌等多种因素有关，金属元素暴露过量也是重要原因之一［2］。铝是地壳中含量最丰富的金属元素，由于其具有良好的理化性质，在日常生活中的使用非常广泛，人类主要通过职业性、医源性和生活性接触铝［3］。……