【摘要】：棉籽餅粕是廣泛應(yīng)用于畜牧生產(chǎn)的飼料原料,棉酚廣泛存在于棉籽植物中,具有生殖毒性及其他毒性作用。為研究棉酚對(duì)雄性小鼠生殖毒性的機(jī)制,選用清潔級(jí)雄性昆明小鼠60只(體重為20.43±1.57 g),飼喂1周后隨機(jī)分成4組:A(對(duì)照組)、B(40 mg/kg)、C(80mg/kg)、D(120 mg/kg),每組15只,自由飲水、采食。對(duì)照組灌胃1%羧甲基纖維素鈉,試驗(yàn)組分別灌胃由1%羧甲基纖維素鈉稀釋的棉酚濃度為40、80、120 mg/kg的懸濁液,連續(xù)灌胃20 d。第21天頸椎脫臼處死小鼠,取睪丸組織用于檢測(cè)。DCFH-DA探針標(biāo)記法測(cè)定睪丸組織ROS的含量,QRT-PCR法檢測(cè)小鼠睪丸組織中各基因mRNA的表達(dá),Western blot法檢測(cè)小鼠睪丸組織中各基因蛋白的表達(dá)。結(jié)果顯示,與對(duì)照組相比,棉酚處理組睪丸組織中ROS含量增加且隨著棉酚劑量的增加而增加,80、120 mg/kg組差異極顯著(P0.01,P0.001)。40、80、120 mg/kg組JNK1 mRNA表達(dá)量顯著升高(P0.05),40、80、120 mg/kg組JNK2 mRNA表達(dá)量極顯著升高(P0.01),80、120 mg/kg組JNK3 mRNA表達(dá)量極顯著升高(P0.001),80、120 mg/kg組MKK4 mRNA表達(dá)量顯著升高(P0.05,P0.001),80、120 mg/kg組FAS和FASL mRNA表達(dá)量極顯著升高(P0.01,P0.001),120 mg/kg組C-MYC mRNA表達(dá)量極顯著升高(P0.001)。其中80mg/kg的棉酚能顯著提高P-MKK4、P-JNK、FAS、FASL蛋白的表達(dá)(P0.05,P0.01),120mg/kg的棉酚能極顯著的提高P-MKK4、P-JNK、FAS、FASL和C-MYC蛋白的表達(dá)(P0.01,P0.001)。結(jié)論,灌胃80、120 mg/kg的棉酚能顯著影響雄性昆明小鼠睪丸組織中ROS-JNK通路的活性。表明棉酚能夠通過(guò)增加睪丸細(xì)胞中內(nèi)源性ROS的含量,激活JNK信號(hào)通路,并促進(jìn)凋亡系統(tǒng)FAS/FASL和原癌基因C-MYC的表達(dá),誘導(dǎo)睪丸細(xì)胞的凋亡,進(jìn)而對(duì)雄性小鼠的生殖系統(tǒng)造成損害,這可能是棉酚抗生育作用的主要機(jī)制之一。
[Abstract]:Cottonseed meal is a kind of feed raw material widely used in animal husbandry. Gossypol widely exists in cottonseed plants and has reproductive toxicity and other toxic effects. In order to study the mechanism of reproductive toxicity of gossypol to male mice, 60 clean male Kunming mice were randomly divided into four groups: A (control group (), B (40 mg/kg), C (80mg/kg) after feeding 20. 43 鹵1. 57 g), for one week. D (120 mg/kg), 15 rats in each group, free drinking water and feeding. The control group was given 1% sodium Carboxymethyl cellulose, and the experimental group was treated with gossypol concentration of 40,80120 mg/kg diluted by 1% sodium Carboxymethyl cellulose for 20 days. On the 21st day, the mice were killed by dislocated cervical vertebrae and used to detect the testicular tissue. The content of ROS in testicular tissue was determined by DCFH-DA probe labeling, and the expression of mRNA in testicular tissue was detected by QRT-PCR. The expression of each gene protein in mouse testicular tissue was detected by Western blot. The results showed that compared with the control group, the content of ROS in testicular tissue of gossypol treatment group increased and increased with the increase of gossypol dose, and the difference was very significant in 80120 mg/kg group (P0.01). The expression of JNK1 mRNA in 80120 mg/kg group was significantly increased (P 0.05), the expression of JNK2 mRNA in 80120 mg/kg group was significantly increased (P 0.01), and the expression of JNK3 mRNA in 80120 mg/kg group was significantly increased (P0.001). The expression of MKK4 mRNA in 80120 mg/kg group was significantly increased (P0.05, P0.001), the expression of FAS and FASL mRNA in 80120 mg/kg group was significantly increased (P0.01, P0.001), and the expression of C-MYC mRNA in 80120 mg/kg group was significantly increased (P0.001). Gossypol in 80mg/kg could significantly increase the expression of P 鈮，

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