4-壬基酚联合双酚A通过抑制Akt/mTOR通路诱导F1代大鼠睾丸细胞自噬和精子发生障碍  

4-nonylphenol combined with bisphenol A induces testicular autophagy and spermatogenic impairment by inhibiting the Akt/m TOR pathway in F1 male offspring rats

作　　者：蔡家星 李桢 代娟[2] 王雪琳 张浩 成金鑫 施筱凡 张玲 石玉琴 CAI Jia-xing;LI Zhen;DAI Juan;WANG Xue-lin;ZHANG Hao;CHENG Jin-xin;SHI Xiao-fan;ZHANG Ling;SHI YU-qin(Key Laboratory of Occupational Hazards Identification and Control,School of Medicine,Wuhan University of Science and Technology,Wuhan,Hubei 430065,China;Department of Chronic Diseases,Wuhan Center for Disease Control and Prevention,Wuhan,Hubei 430015,China;Department of Obstetrics and Gynecology,Xiangy-ang First Hospital,Hubei Medical College,Xiangyang,Hubei 441000,China)

机构地区：[1]武汉科技大学医学院公共卫生学院职业危害识别与控制湖北省重点实验室,湖北武汉430065 [2]武汉市疾病预防控制中心慢病所,湖北武汉430015 [3]湖北医药学院附属襄阳市第一人医院妇产科,湖北襄阳441000

出　　处：《中华男科学杂志》2022年第3期195-202,共8页National Journal of Andrology

基　　金：湖北省卫生健康委员会联合基金面上项目(WJ2019H255);职业危害识别与控制湖北省重点实验室开放基金(OHIC2019G03);湖北省高等学校优秀中青年科技创新团队项目(T2020003)。

摘　　要：目的:研究F0代4-壬基酚(4-NP)和双酚A(BPA)暴露对F1代大鼠睾丸细胞自噬及Akt/mTOR通路关键基因表达的影响。方法:采用2×2析因设计,将F0代雌雄鼠随机分为对照组(植物油)、BPA组(0.5 mg/kg BPA)、4-NP组(5 mg/kg 4-NP)、BPA+4-NP组(0.5 mg/kg BPA+5 mg/kg 4-NP),连续隔天灌胃染毒30 d后,同组雌雄鼠3∶1合笼受孕,雌鼠怀孕后继续隔天灌胃染毒直至分娩。F1代子鼠从哺乳期至60日龄,期间均未给予4-NP和BPA暴露,随后处死大鼠。对60日龄F1代子鼠精子进行计数,观察睾丸组织形态学和自噬的组间变化,检测睾丸组织Akt、mTOR、4EBP1和p70S6K mRNA的表达水平及LC3蛋白的表达水平。结果:与对照组比较,BPA组和BPA+4-NP组处理的F1代大鼠精子数量明显下降(P<0.05),BPA和4-NP对F1代大鼠精子数量的影响无交互作用(P_(交互)>0.05);NP、BPA和BPA+4-NP组LC3-II/LC3-I比值上升,BPA和4-NP对LC3-II/LC3-I比值的影响有交互作用,睾丸生精小管内细胞排列松散,管腔内精子数量有降低趋势,胞内自噬泡数量有增加的趋势;4-NP组p70S6K、BPA组mTOR、NP+BPA组Akt mRNA表达明显下降(P<0.05),各染毒组4EBP1 mRNA表达明显升高(P<0.05),BPA和4-NP对Akt和4EBP1 mRNA表达的影响有交互作用(P_(交互)<0.05)。结论:F0代低浓度4-NP和BPA长期染毒对F1代大鼠睾丸功能有损伤作用,此过程与细胞自噬及Akt/mTOR通路改变密切相关。Objective: To study the impacts of exposure of F0 generation rats to 4-nonylphenol(4-NP) and bisphenal A(BPA) on the autophagy of testicular cells and key gene expressions of the Akt/mTOR pathway in the F1 generation offspring rats. Methods: Using a 2×2 factorial design, F0 female and male rats were randomly assigned to receive intragastrically sigma vegetable oil(the control), BPA at 0.5 mg/kg, 4-NP at 5 mg/kg, and BPA+4-NP both at 0.5 mg/kg, respectively, qd alt for 30 days. Then the rats in each group were mated at a ratio of 1∶3. After pregnancy, the female rats continued the above intragastrical administration till delivery. The F1 generation male offspring rats were killed at 60 postnatal days, and their testes harvested for sperm count, observation of the morphological and autophagic changes of the testis, and determination of the relative mRNA expression levels of Akt, mTOR, 4 EBP1 and p70 S6 K and protein expression levels of LC3-I and LC3-II. Results: The sperm count of the F1 generation male offspring rats was markedly decreased in the BPA and BPA+4-NP groups compared with that in the control(P < 0.05) but no significant interactive effect was observed between BPA and 4-NP(P > 0.05). The ratio of LC3-II/LC3-I was remarkably increased in the 4-NP, BPA and BPA+4-NP groups in comparison with that in the control(P < 0.05), and a significant interactive effect was shown between BPA and 4-NP(P < 0.05). The cells in the seminiferous tubules of the rats in the 4-NP, BPA and BPA+4-NP groups were loosely arranged, with a decreased count of sperm and increased number of autophagic vacuoles. Significant down-regulation was observed in the relative mRNA expression of p70 S6 K in the 4-NP group, as well as in that of mTOR in the BPA group and Akt in the BPA+4-NP group(all P < 0.05). A remarkably up-regulated expression of 4 EBP1 mRNA was found in all the three intervention groups(P < 0.05), with a significant interactive effect between BPA and 4-NP on the expressions of Akt and 4 EBP1 mRNA(P < 0.05). Conclusion:

关 键 词：4-壬基酚 双酚A 自噬 Akt/mTOR通路 大鼠 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学]