氧化应激对公鸡睾丸组织结构及睾酮合成相关基因表达的影响  

Effects of Oxidative Stress on Testicular Structure and Expression of Genes Related to Testosterone Synthesis in Roosters(Gallus gallus)

作　　者：吴冰燕 曹原 刘思丽 刘亚美 丁香 凌义强 胡威 方满新 刘犇[1,2,3] 王小月 WU Bing-Yan;CAO Yuan;LIU Si-Li;LIU Ya-Mei;DING Xiang;LING Yi-Qiang;HU Wei;FANG Man-Xin;LIU Ben;WANG Xiao-Yue(College of Life Science and Resources and Environment,Yichun University,Yichun 336000,China;Jiangxi Lvke Agriculture and Animal Husbandry Technology Co.,Ltd.,Yichun 336000,China;Engineering Technology Research Center of Jiangxi Universities and Colleges for Selenium Agriculture,Yichun 336000,China)

机构地区：[1]宜春学院生命科学与资源环境学院,宜春336000 [2]江西绿科农牧科技有限公司,宜春336000 [3]江西省高等学校硒农业工程技术研究中心,宜春336000

出　　处：《农业生物技术学报》2024年第3期605-616,共12页Journal of Agricultural Biotechnology

基　　金：江西省教育厅科学技术研究项目(GJJ2201729);宜春学院科研启动经费(210-3360119068);宜春学院大学生创新创业训练计划项目(X202110417416)。

摘　　要：禽类生长繁殖过程中易受各种应激因素的影响,其中以氧化应激最为突出。为探究氧化应激对鸡(Gallus gallus)睾丸组织结构及睾酮合成相关基因表达的影响,本研究利用敌草快(diquat)作为氧化应激诱导剂,将60只180日龄的健康康乐黄公鸡随机平均分成4组:对照组、敌草快5 mg/kg组、敌草快15 mg/kg组和敌草快20 mg/kg组。待21 d处理结束后采集鸡睾丸组织样品,利用qRT-PCR技术和抗氧化功能检测试剂盒分别检测氧化应激相关基因核因子红细胞2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2)、半氧合酶1(hemeoxygenase-1,HO-1)、谷胱甘肽过氧化物酶1(glutathione peroxidase 1,Gpx1)的表达水平和过氧化氢酶(catalase,CAT)、总超氧化物歧化酶(total superoxide dismutase,T-SOD)的活性及丙二醛(malondialdehyde,MDA)的浓度,确定氧化应激发生情况;利用苏木精-伊红染色法(hematoxylin-eosin staining,HE staining)检测氧化应激对睾丸组织结构的影响,利用Western blot技术分别检测细胞凋亡相关蛋白B细胞淋巴瘤-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)和半胱氨酸蛋白酶3(Caspase3)的表达量变化,通过qRT-PCR和Western blot技术分别检测睾酮合成相关基因类固醇激素合成急性调节蛋白(steroidogenic acute regulatory protein,StAR)、细胞色素P450胆固醇侧链裂解酶(cytochrome P450 family 11 subfamily A member 1,Cyp11a1)和3β-羟基类固醇脱氢酶1(3β-hydroxysteroid dehydrogenase 1,Hsd3b1)的mRNA和蛋白表达量变化。结果显示,与对照组相比,不同剂量敌草快组睾丸中氧化应激相关分子HO-1、Nrf2和Gpx1的mRNA水平显著下调(P<0.05),CAT和T-SOD酶活性显著降低(P<0.05),MDA浓度显著升高(P<0.05);HE染色结果表明,睾丸经敌草快诱导作用后结构受到损伤,曲精小管中的精子发生受到明显抑制,随着敌草快处理浓度的升高,曲精小管中的生殖细胞发生严重凋亡;Bax和Caspase3的蛋�During the process of growth and reproduction,poultry is susceptible to various stress factors,the most prominent of which is oxidative stress.To investigate the effects of oxidative stress on the testicular structure and expression of genes related to testosterone synthesis in chickens(Gallus gallus),60 healthy Kangle yellow roosters aged 180 d were randomly divided into 4 groups:Control group,diquat 5 mg/kg group,diquat 15 mg/kg group and diquat 20 mg/kg group.Testicular tissue samples were collected after 21 d of treatment.qRT-PCR was used to identify the presence of oxidative stress by measuring the expressions of nuclear factor erythroid 2-related factor 2(Nrf2),hemeoxygenase-1(HO-1)and glutathione peroxidase 1(Gpx1),and Antioxidant Function Assay Kit was used to detect the enzyme activities of catalase(CAT),total superoxide dismutase(T-SOD)and the concentration of malondialdehyde(MDA).Hematoxylin-eosin staining(HE staining)was used to analyze the effect of oxidative stress on testicular tissue structure.The relative protein expressions of apoptosis-related B-cell lymphoma-2 associated X protein(Bax),B-cell lymphoma-2(Bcl-2)and Caspase3 were detected by using Western blot.The relative mRNA and protein expressions of steroidogenic acute regulatory protein(StAR),cytochrome P450 family 11 subfamily A member 1(Cyp11a1)and 3β-hydroxysteroid dehydrogenase(Hsd3b1),which were involved in testosterone biosynthesis,were detected by using qRT-PCR and Western blot,respectively.The results showed that the mRNA level of oxidative stress-related genes HO-1,Nrf2 and Gpx1 were significantly down-regulated in the testes of the different doses diquat groups when compared to the control group(P<0.05).CAT and T-SOD activity were significantly decreased(P<0.05),while MDA concentration was significantly increased(P<0.05).After diquat induction,the structure of chicken testes was damaged,and spermatogenesis in seminiferous tubules was significantly inhibited with severe cell apoptosis,according to HE staining.The protein expressio

关 键 词：氧化应激 公鸡 睾丸 细胞凋亡 睾酮合成相关基因 

分 类 号：S856.9[农业科学—临床兽医学]