棉酚对小鼠睾丸组织中ROS-JNK通路的影响
发布时间：2019-05-24 20:34
【摘要】：棉籽饼粕是广泛应用于畜牧生产的饲料原料,棉酚广泛存在于棉籽植物中,具有生殖毒性及其他毒性作用。为研究棉酚对雄性小鼠生殖毒性的机制,选用清洁级雄性昆明小鼠60只(体重为20.43±1.57 g),饲喂1周后随机分成4组:A(对照组)、B(40 mg/kg)、C(80mg/kg)、D(120 mg/kg),每组15只,自由饮水、采食。对照组灌胃1%羧甲基纤维素钠,试验组分别灌胃由1%羧甲基纤维素钠稀释的棉酚浓度为40、80、120 mg/kg的悬浊液,连续灌胃20 d。第21天颈椎脱臼处死小鼠,取睾丸组织用于检测。DCFH-DA探针标记法测定睾丸组织ROS的含量,QRT-PCR法检测小鼠睾丸组织中各基因mRNA的表达,Western blot法检测小鼠睾丸组织中各基因蛋白的表达。结果显示,与对照组相比,棉酚处理组睾丸组织中ROS含量增加且随着棉酚剂量的增加而增加,80、120 mg/kg组差异极显著(P0.01,P0.001)。40、80、120 mg/kg组JNK1 mRNA表达量显著升高(P0.05),40、80、120 mg/kg组JNK2 mRNA表达量极显著升高(P0.01),80、120 mg/kg组JNK3 mRNA表达量极显著升高(P0.001),80、120 mg/kg组MKK4 mRNA表达量显著升高(P0.05,P0.001),80、120 mg/kg组FAS和FASL mRNA表达量极显著升高(P0.01,P0.001),120 mg/kg组C-MYC mRNA表达量极显著升高(P0.001)。其中80mg/kg的棉酚能显著提高P-MKK4、P-JNK、FAS、FASL蛋白的表达(P0.05,P0.01),120mg/kg的棉酚能极显著的提高P-MKK4、P-JNK、FAS、FASL和C-MYC蛋白的表达(P0.01,P0.001)。结论,灌胃80、120 mg/kg的棉酚能显著影响雄性昆明小鼠睾丸组织中ROS-JNK通路的活性。表明棉酚能够通过增加睾丸细胞中内源性ROS的含量,激活JNK信号通路,并促进凋亡系统FAS/FASL和原癌基因C-MYC的表达,诱导睾丸细胞的凋亡,进而对雄性小鼠的生殖系统造成损害,这可能是棉酚抗生育作用的主要机制之一。
[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 卤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，
本文编号：2485163
[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 卤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，
本文编号：2485163
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