钼致小鼠睾丸组织氧化损伤及对PI3K/AKT信号通路的影响

Effects of molybdenum on oxidative damage and PI3K/AKT signaling pathway in mice testis

为探索钼致雄性小鼠的睾丸组织的氧化损伤及其对PI3K/AKT信号通路的影响,本试验选取4周龄清洁级雄性健康昆明系小鼠60只,随机分为Ⅰ、Ⅱ、Ⅲ、Ⅳ组,分别在饮水中添加0、100、200、400 mg/L的钼,基础日粮相同,自由饮水,试验周期90 d。试验结束后剖杀并采集睾丸。比色法检测睾丸组织SOD、T-AOC、MDA、LDH活性或含量;RT-PCR和Western-blotting测定PI3K/AKT信号通路上PI3K、PTEN、AKT、Caspase-9、Bcl-2、Bax和Bad的m RNA和蛋白的表达情况。结果显示,与Ⅰ组相比,Ⅱ、Ⅲ、Ⅳ组SOD和T-AOC活性极显著下降(P<0.01),Ⅱ组LDH活性显著下降(P<0.05),Ⅲ、Ⅳ组LDH活性极显著上升(P<0.01),Ⅱ、Ⅲ、Ⅳ组MDA含量极显著升高(P<0.01)。与Ⅰ组相比,Ⅱ组和Ⅲ组PI3K、AKT、Bcl-2、Bad和Bax的m RNA表达量显著升高(P<0.05),Ⅱ组PTEN、Caspase-9的m RNA表达量显著下降(P<0.05),Ⅲ组Caspase-9的m RNA表达量显著升高(P<0.05);Ⅳ组PI3K、AKT、Caspase-9、Bcl-2、Bad和Bax的m RNA表达量显著下降(P<0.05),PTEN的m RNA表达量显著升高(P<0.05)。与Ⅰ组相比,Ⅱ组和Ⅲ组PI3K、AKT、P-AKT、Caspase-9、Bcl-2和Bax蛋白表达量显著升高(P<0.05),Ⅱ组PTEN蛋白表达量显著下降(P<0.05);Ⅳ组PI3K、PTEN、AKT和Bax蛋白表达量显著升高(P<0.05),Bcl-2和Bad蛋白表达量显著下降(P<0.05)。本试验所设剂量钼均能导致小鼠睾丸组织氧化损伤,同时通过激活PI3K/AKT信号通路上相关信号因子m RNA和蛋白表达对雄性小鼠生殖系统产生影响。

The aim of this study is to investigate the effect of oxidative damage and PI3 K/AKT signaling pathway induced by molybdenum on testis tissue of male mice.60 4-week-old clean grade healthy Kunming male mice were selected in this experiment.They were randomly divided into four groups(groupⅠ,groupⅡ,groupⅢ and groupⅣ).0,100,200,400 mg/L sodium molybdenum were added to groupⅠ,groupⅡ,group Ⅲ and group Ⅳ separately in drinking water.The rats had the same basic diet and free drinking water.The experiment lasted for 90 days.At the end of the experimental period,the testes were collected from all animals.Colorimetric assay was used for the detection of SOD,T-AOC,MDA and LDH activities or contents in testicular tissue.The m RNA and protein expressions of PI3 K,PTEN,AKT,Caspase-9,Bcl-2,Bax and Bad in PI3 K/AKT signaling pathway were determined by RT-PCR and Western-blotting.In result,compared with groupⅠ,the activities of SOD and T-AOC decreased significantly in groupⅡ,Ⅲ and Ⅳ(P<0.01),the activity of LDH in groupⅡ decreased significantly(P<0.05),but the activity of LDH in group Ⅲ andⅣ increased significantly(P<0.01),the MDA content in group Ⅱ,Ⅲ and Ⅳ increased significantly(P<0.01).Compared with groupⅠ,the m RNA expression levels of PI3 K,AKT,Bcl-2,Bad and Bax in group Ⅱand Ⅲ increased significantly(P<0.05),and the m RNA expressions of PTEN and Caspase-9 in group Ⅱ decreased significantly(P<0.05).The m RNA expression of Caspase-9 in group Ⅲ increased significantly(P <0.05).The m RNA expression of PI3 K,AKT,Caspase-9,Bcl-2,Bad and Bax decreased significantly in group Ⅳ(P<0.05),and the m RNA expression of PTEN increased significantly(P<0.05).Compared with groupⅠ,the expression of PI3 K,AKT,P-AKT,Caspase-9,Bcl-2 and Bax protein in group Ⅱ and group Ⅲ increased significantly(P<0.05),and the expression of PTEN protein decreased in groupⅡ significantly(P<0.05).The expression of PI3 K,PTEN,AKT and Bax protein increased significantly(P<0.05),and the expression of Bcl-2 and Bad protein decreased significantly(P<0.05).The above-mentioned results showed that different concentrations of molybdenum can cause oxidative damage in testis of male mice.At the same time,molybdenum can affect male reproductive system by regulating the expression of related factors m RNA and protein in PI3 K/AKT signaling pathway in mice.