【摘要】：[目的]研究二硫化碳(carbon disulfide,CS2)對大鼠睪丸生殖細(xì)胞凋亡線粒體通路的影響及其可能的分子機(jī)制,同時(shí)觀察環(huán)孢素A(cyclosporin A,Cs A)對細(xì)胞凋亡的干預(yù)作用。[方法]選取清潔級雄性SD大鼠48只并隨機(jī)分為6組:對照組、3個CS2染毒組(50、250、1 250 mg/m3)、干預(yù)組[CS2(1 250 mg/m3)+Cs A(12.5 mg/kg)]和Cs A(12.5 mg/kg)組。靜式吸入染毒,每天2 h,每周5 d,共10周,對照組僅吸入新鮮空氣,Cs A則依據(jù)大鼠體重并按相應(yīng)比例先溶解在牛奶中然后灌胃。染毒結(jié)束后取睪丸組織做HE染色,觀察其形態(tài)學(xué)改變;TUNEL熒光法檢測大鼠睪丸組織細(xì)胞凋亡情況;Western Blot法檢測各組細(xì)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的表達(dá)含量。[結(jié)果]隨著CS2染毒濃度的增加,HE染色大鼠睪丸組織形態(tài)損傷情況不斷加劇,中、高濃度組的凋亡指數(shù)高于對照組(P0.05),Cs A干預(yù)對大鼠睪丸組織形態(tài)損傷有一定緩解;與高濃度CS2染毒組相比,干預(yù)組的凋亡指數(shù)明顯下降(P0.05),但仍高于Cs A組(P0.05)。CS2染毒組Cyt C、Bad蛋白含量與對照組比較有所增加,而Procaspase-9、Procaspase-3和Bcl-x L蛋白含量則有下降的趨勢(P0.05)。Cs A干預(yù)后與高濃度CS2染毒組比較,干預(yù)組Cyt C蛋白含量下降(P0.05),Bcl-x L蛋白含量則提高(P0.05);Procaspase-9、Procaspase-3和Bad蛋白含量均有不同程度的變化,但與CS2高濃度組相比差異無統(tǒng)計(jì)學(xué)意義。[結(jié)論]CS2染毒可影響細(xì)胞凋亡線粒體通路繼而誘導(dǎo)大鼠睪丸生殖細(xì)胞凋亡,而細(xì)胞凋亡線粒體通路相關(guān)蛋白的表達(dá)可能與Cs A的干預(yù)有關(guān)。
[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.
【作者單位】： 華中科技大學(xué)同濟(jì)醫(yī)學(xué)院公共衛(wèi)生學(xué)院勞動衛(wèi)生與環(huán)境衛(wèi)生學(xué)系;
【基金】：國家自然科學(xué)基金項(xiàng)目(編號:81172637)
【分類號】：R114

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