王为, 郭寅生, 张振, 董煜, 黄晓彧, 周义军, 陈国元. 二硫化碳对大鼠睾丸组织细胞凋亡线粒体通路的影响及环孢素A的干预作用[J]. 环境与职业医学, 2015, 32(3): 199-205. DOI: 10.13213/j.cnki.jeom.2015.14371
引用本文: 王为, 郭寅生, 张振, 董煜, 黄晓彧, 周义军, 陈国元. 二硫化碳对大鼠睾丸组织细胞凋亡线粒体通路的影响及环孢素A的干预作用[J]. 环境与职业医学, 2015, 32(3): 199-205. DOI: 10.13213/j.cnki.jeom.2015.14371
WANG Wei , GUO Yin-sheng , ZHANG Zhen , DONG Yu , HUANG Xiao-yu , ZHOU Yijun , CHEN Guo-yuan . Effects of Carbon Disulfide on Testicular Apoptosis via Mitochondrial Pathways and the Intervention of Cyclosporin A[J]. Journal of Environmental and Occupational Medicine, 2015, 32(3): 199-205. DOI: 10.13213/j.cnki.jeom.2015.14371
Citation: WANG Wei , GUO Yin-sheng , ZHANG Zhen , DONG Yu , HUANG Xiao-yu , ZHOU Yijun , CHEN Guo-yuan . Effects of Carbon Disulfide on Testicular Apoptosis via Mitochondrial Pathways and the Intervention of Cyclosporin A[J]. Journal of Environmental and Occupational Medicine, 2015, 32(3): 199-205. DOI: 10.13213/j.cnki.jeom.2015.14371

二硫化碳对大鼠睾丸组织细胞凋亡线粒体通路的影响及环孢素A的干预作用

Effects of Carbon Disulfide on Testicular Apoptosis via Mitochondrial Pathways and the Intervention of Cyclosporin A

  • 摘要: 目的 研究二硫化碳(carbon disulfide, CS2)对大鼠睾丸生殖细胞凋亡线粒体通路的影响及其可能的分子机制,同时观察环孢素A(cyclosporin A, CsA)对细胞凋亡的干预作用。

    方法 选取清洁级雄性SD大鼠48只并随机分为6组:对照组、3个CS2染毒组(50、250、1 250 mg/m3)、干预组CS2(1 250 mg/m3)+CsA(12.5 mg/kg)和CsA(12.5 mg/kg)组。静式吸入染毒,每天2 h,每周5 d,共10周,对照组仅吸入新鲜空气, CsA则依据大鼠体重并按相应比例先溶解在牛奶中然后灌胃。染毒结束后取睾丸组织做HE染色,观察其形态学改变; TUNEL荧光法检测大鼠睾丸组织细胞凋亡情况; Western Blot法检测各组细胞色素C(Cyt C)、半胱氨酸天冬氨酸酶原9(Procaspase-9)、Procaspase-3、Bcl-xL抗凋亡和促凋亡蛋白Bad的表达含量。

    结果 随着CS2染毒浓度的增加, HE染色大鼠睾丸组织形态损伤情况不断加剧,中、高浓度组的凋亡指数高于对照组(P < 0.05), CsA干预对大鼠睾丸组织形态损伤有一定缓解;与高浓度CS2染毒组相比,干预组的凋亡指数明显下降(P<0.05),但仍高于CsA组(P<0.05)。CS2染毒组Cyt C、Bad蛋白含量与对照组比较有所增加,而Procaspase-9、Procaspase-3和Bcl-xL蛋白含量则有下降的趋势(P < 0.05)。CsA干预后与高浓度CS2染毒组比较,干预组Cyt C蛋白含量下降(P < 0.05), Bcl-xL蛋白含量则提高(P < 0.05); Procaspase-9、Procaspase-3和Bad蛋白含量均有不同程度的变化,但与CS2高浓度组相比差异无统计学意义。

    结论 CS2染毒可影响细胞凋亡线粒体通路继而诱导大鼠睾丸生殖细胞凋亡,而细胞凋亡线粒体通路相关蛋白的表达可能与CsA的干预有关。

     

    Abstract: Objective To investigate the effects of carbon disulfide (CS2) on testicular apoptosis via mitochondrial pathways, explore its possible molecular mechanisms, and observe the intervention of cyclosporin A (CsA) on cell apoptosis.

    Methods Forty-eight SD male rats were randomly divided into six groups. Three groups were exposed to CS2 at designed concentrations (50, 250, 1 250 mg/m3) by inhalation 2 h/d and 5 d/week for 10 weeks. Fresh air was inhaled only by rats of the control group. The other two groups were treated with CS2 (1250mg/m3) plus CsA (12.5mg/kg) and CsA (12.5mg/kg) alone respectively. CsA was prepared before gavage to the designed solutions by milk as solvent according to rat body weights. The histopathological changes in rats' testes were observed after HE staining and apoptosis by TUNEL. Western Blot was applied to detect the expression of Cyt C, Procaspase-9, Procaspase-3, Bcl-xL, and Bad protein in each group.

    Results The histopathological injuries of rats' testes stained by HE were aggravated with the increasing of CS2 concentration. The apoptosis indices of the middle and high concentration groups were higher than that of the control group (P < 0.05). After treated with CsA, the histopathological injuries of rats' testes were alleviated, and also the apoptosis index was declined significantly compared with the high concentration group (P < 0.05) but higher than that of the CsA only group (P < 0.05). Compared with the control group, the expressions of Cyt C and Bad protein were increased, whereas the expressions of Procaspase-9, Procaspase-3, and Bcl-xL were lowered in different concentration groups (P < 0.05). In the CS2 plus CsA group, the expression of Cyt C was decreased and the expression of Bcl-xL was increased compared with the high concentration group (both P < 0.05). Varied changes were observed in the expressions of Procaspase-9 and Bad protein in the CS2 plus CsA group; however, there was no statistical difference in comparison with the high concentration group.

    Conclusion CS2 could induce testicular apoptosis through mitochondrial pathways. Meanwhile, CsA might regulate the expression of proteins related to mitochondrial pathways.

     

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