硒对铅致小鼠睾丸间质细胞凋亡的影响

doi:10.16431/j.cnki.1671-7236.2023.04.012

摘要/Abstract

摘要： 【目的】探究硒对铅致小鼠睾丸间质细胞凋亡的影响。【方法】以小鼠睾丸间质细胞为研究对象,分为对照组、硒组、铅组和硒＋铅组:硒组在细胞培养液中添加2 μmol/L亚硒酸钠;铅组在细胞培养液中添加40 μmol/L醋酸铅;硒＋铅组在细胞培养液中联合添加2 μmol/L亚硒酸钠＋40 μmol/L醋酸铅,体外培养24 h。采用MTT法检测细胞增殖情况;检测细胞内谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量;使用化学荧光法测定活性氧(ROS)水平;运用实时荧光定量PCR和Western blotting分别检测凋亡相关基因天冬氨酸半胱氨酸酶-3(Caspase3)、Caspase8、B淋巴细胞瘤-2(Bcl-2)和Bcl-2相关X蛋白(Bax)的mRNA和蛋白表达水平。【结果】与对照组相比,铅组细胞增殖显著受到抑制(P＜0.05),MDA含量和ROS水平显著上升(P＜0.05),SOD和GSH-Px活性显著下降(P＜0.05),同时,铅显著上调了Caspase3、Caspase8、Bax的mRNA和蛋白表达水平(P＜0.05),显著下调了Bcl-2的mRNA和蛋白表达水平(P＜0.05);硒组小鼠睾丸间质细胞增殖活性显著提高(P＜0.05)。与铅组相比,硒＋铅组细胞中MDA含量和ROS水平显著下降(P＜0.05),SOD和GSH-Px活性显著升高(P＜0.05),且介于对照组和铅组之间;Caspase3、Caspase8、Bax的mRNA和蛋白表达水平显著下降(P＜0.05),Bcl-2 mRNA和蛋白表达水平则显著上升(P＜0.05),均介于铅组和对照组之间。【结论】铅能抑制小鼠睾丸间质细胞增殖活力,诱导细胞氧化应激和凋亡;补充适量硒可以促进细胞增殖,且通过提高抗氧化酶活性缓解铅造成的细胞氧化应激和凋亡。

关键词: 硒; 铅; 小鼠; 睾丸间质细胞; 凋亡

Abstract: 【Objective】 The aim of this study was to investigate the effects of selenium on lead-induced apoptosis in mouse Leydig cells in vitro.【Method】 Mouse Leydig cells were divided into control,selenium,lead and selenium＋lead groups.2 μmol/L sodium selenite was added into the cell culture medium of selenium group,40 μmol/L lead acetate was added into the cell culture medium of lead group,2 μmol/L sodium selenite and 40 μmol/L lead acetate were added into the cell culture medium of selenium＋lead group.The cells were cultured for 24 h in vitro.Cell proliferation was detected by MTT assay.The activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in cells were detected by biochemical analysis.Reactive oxygen species (ROS) level was measured by chemical fluorescence.Real time quantitative PCR and Western blotting were used to detect the mRNA and protein expression levels of cysteinyl aspartate specific proteinase 3 (Caspase3),cysteinyl aspartate specific proteinase 8 (Caspase8),B-cell lymphoma-2 (Bcl-2) and Bcl2-associated X (Bax).【Result】 Compared with control group, cell proliferation was significantly inhibited in lead group (P＜0.05),the content of MDA and ROS level were significantly increased (P＜0.05),and the activities of SOD and GSH-Px were significantly decreased (P＜0.05).At the same time,lead significantly up-regulated the mRNA and protein levels of Caspase3,Caspase8 and Bax (P＜0.05),but down-regulated the mRNA and protein levels of Bcl-2 (P＜0.05).The proliferative activity of Leydig cells in selenium group was significantly increased than that in control group (P＜0.05).Compared with lead group,the content of MDA and ROS level in the cells of the selenium＋lead group were significantly decreased,and the activities of SOD and GSH-Px were significantly increased,which were between control and lead groups (P＜0.05).The mRNA and protein expressions of Caspase3,Caspase8,and Bax in selenium＋lead group were significantly lower than those in lead group (P＜0.05),while Bcl-2 showed an opposite trend (P＜0.05).【Conclusion】 Lead could inhibit the proliferative activity of mouse Leydig cells in vitro and induce oxidative stress and apoptosis.Supplementing appropriate amount of selenium could promote cell proliferation,alleviate lead-induced cell oxidative stress and apoptosis by increasing the activity of antioxidant enzymes.

Key words: selenium; lead; mouse; Leydig cells; apoptosis

中图分类号:

S814

郭雅茹, 李可欣, 王丽, 刘锦钰, 石磊, 任有蛇. 硒对铅致小鼠睾丸间质细胞凋亡的影响[J]. 中国畜牧兽医, 2023, 50(4): 1404-1411.

GUO Yaru, LI Kexin, WANG Li, LIU Jinyu, SHI Lei, REN Youshe. Effects of Selenium on Lead-Induced Apoptosis of Mouse Leydig Cells in vitro[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1404-1411.

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