乙肝病毒S蛋白暴露时间对人精子胞膜完整性及功能的影响

Effects of hepatitis B virus S protein exposuring time on human sperm membrane integrity and function

目的:探讨乙肝病毒S蛋白(HBs)与人精子体外共孵育不同时间后人精子胞膜完整性及功能的变化情况。方法:人精子与25μg/m L HBs共孵育0、1、2、3、4 h后,酶标仪检测精子细胞内丙二醛(MDA)含量;Annexin V-FITC/PI双染法检测HBs对精子早期凋亡指标磷脂酰丝氨酸外翻情况,流式细胞术检测精子细胞内活性氧(ROS)、半胱氨酸蛋白酶-3,-8,-9(Caspase-3,-8,-9)活化,TUNEL结合流式细胞术检测精子DNA的损伤。结果:人精子与25μg/m L HBs共孵育后,MDA水平、ROS阳性精子、Annexin V+/PI-精子、Caspase-3,-8,-9阳性精子比率、TUNEL阳性精子比率显著升高,且Annexin V+/PI-精子、Caspase-8,-9阳性精子比率、TUNEL阳性精子比率的升高与HBs呈时间依赖趋势。结论:HBs蛋白与人精子共孵育时间增长可加剧对人精子功能的损伤。

AIM： To explore the relationship between hepatitis B virus S protein（ HBs） exposure time and human sperm membrane integrity and function in human sperm cells. METHODS： After the co-incubation of sperm cells with 25 μg / m L HBs for0 h,1 h,2 h,3 h,4 h,the Aldetect（ MDA-Specific）Lipid Peroxidation（ LP） level was measured by microplatereader; the reactive oxygen species（ ROS）production and PS externalization of membrane,Caspase-3,-8,-9 activation and DNA fragmentation of the human sperm were assessed by flow cytometry. RESULTS： After incubation,the levels of ROS,MDA and the average rates of annexin V-positive /propidium iodide（ PI）-negative sperm,Caspase-3,-8,-9 positive sperm and TUNEL-positive sperm were significantly increased in the test groups as compared with those in the control group（ P〈0. 05,P〈 0. 01）,and HBs increased the annexin V-positive /PI-negative sperm, Caspases-8,-9 positive sperm and TUNEL-positive sperm in a time-dependent manner. CONCLUSION： HBs could aggravate the dysfunction of human sperm in a time-dependent manner.