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

发布时间：2018-10-13 11:05

【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,Cs A)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)+Cs A(12.5 mg/kg)]和Cs A(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,Cs A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-x L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(P0.05),Cs A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(P0.05),但仍高于Cs A组(P0.05)。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-x L蛋白含量则有下降的趋势(P0.05)。Cs A干预后与高浓度CS2染毒组比较,干预组Cyt C蛋白含量下降(P0.05),Bcl-x L蛋白含量则提高(P0.05);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Cs A的干预有关。
[Abstract]:[objective] to study the effect of carbon disulfide (carbon disulfide,CS2) on apoptosis of rat germ cells and its possible molecular mechanism, and to observe the effect of cyclosporine (A (cyclosporin) on apoptosis. [methods] Forty-eight clean grade male SD rats were randomly divided into 6 groups: control group, 3 CS2 exposed groups (50 mg/m3, 250 mg/m3) Cs A (12.5 mg/kg) and Cs A (12.5 mg/kg group. The rats in the control group were inhaled fresh air, Cs A for 10 weeks, 2 hours a day, 5 days a week, and then dissolved in milk according to the weight of the rats and then gavaged according to the corresponding proportion. At the end of the poisoning, the testis were stained with HE. TUNEL fluorescence assay was used to detect apoptosis of rat testis and; Western Blot method was used to detect the expression of cytochrome C (Cyt C), cysteine aspartate proenzyme 9 (Procaspase-9), Procaspase-3,Bcl-x L anti-apoptosis protein and pro-apoptotic protein Bad. [results] with the increase of CS2 concentration, the morphological damage of testis in HE stained rats was aggravated. The apoptotic index in the middle and high concentration group was higher than that in the control group (P0.05). The apoptotic index of the intervention group was significantly lower than that of the high concentration CS2 group (P0.05), but still higher than that of the Cs A group (P0.05). The content of Cyt Cnbad protein in the CS2 group was higher than that in the control group. However, the content of Procaspase-9,Procaspase-3 and Bcl-x L protein decreased (P0.05) compared with the high concentration CS2 group, the content of Cyt C protein decreased (P0.05), the content of Bcl-x L protein increased (P0.05), the content of Procaspase-9,Procaspase-3 and Bad protein changed to some extent, compared with the high concentration of CS2 group, the content of Cyt C protein decreased in the intervention group (P0.05), and the content of Bcl-x L protein increased in the intervention group (P0.05). However, there was no significant difference compared with the high concentration CS2 group. [conclusion] CS2 exposure can affect apoptosis mitochondria pathway and induce germ cell apoptosis in rat testis, and the expression of apoptotic mitochondrial pathway related protein may be related to the intervention of Cs A.
【作者单位】：华中科技大学同济医学院公共卫生学院劳动卫生与环境卫生学系;
【基金】：国家自然科学基金项目(编号:81172637)
【分类号】：R114

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