乙肝病毒S蛋白对人精子氧化应激的影响

doi:10.3969/j.issn.1004-616x.2015.03.009

癌变·畸变·突变 ›› 2015, Vol. 27 ›› Issue (3): 202-206.doi: 10.3969/j.issn.1004-616x.2015.03.009

乙肝病毒S蛋白对人精子氧化应激的影响

康祥锦¹, 丁悦¹, 杨洁¹, 杜红姿¹, 刘见桥¹, 黄天华²

1. 广州医科大学附属第三医院生殖医学中心, 广东省生殖医学重点实验室, 广东省产科重大疾病重点实验室, 广东省普通高校生殖与遗传重点实验室, 广东广州 510150;
2. 汕头大学医学院生殖医学研究中心, 广东汕头 515041

收稿日期:2014-10-16 修回日期:2015-03-02 出版日期:2015-05-30 发布日期:2015-05-30
通讯作者: 黄天华,E-mail:thhuang@stu.edu.cn E-mail:thhuang@stu.edu.cn
作者简介:康祥锦,E-mail:kangxiangjin@163.com
基金资助:
国家自然科学基金资助项目(81401254);广东省人口和计划生育委员会资助项目(20133064);广州市荔湾区科技和信息化局资助项目(20131215083);广州市教育局资助项目(1201430205)

Effects of hepatitis B virus S protein exposure on human sperm oxidative stress

KANG Xiangjin¹, DING Yue¹, YANG Jie¹, DU Hongzi¹, LIU Jianqiao¹, HUANG Tianhua²

1. Center for Reproductive Medicine, Third Affiliated Hospital of Guangzhou Medical University, Key Laboratory for Reproductive Medicine of Guangdong Province, Key Laboratory for Major Obstetric Diseases of Guangdong Province, Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, Guangzhou 510150;
2. Research Center for Reproductive Medicine, Shantou University Medical College, Shantou 515041, Guangdong, China

Received:2014-10-16 Revised:2015-03-02 Online:2015-05-30 Published:2015-05-30
About author:10.3969/j.issn.1004-616x.2015.03.009

摘要/Abstract

摘要：

目的:乙肝病毒(HBV)可通过血睾屏障将病毒DNA整合到男性生殖细胞的基因组中,诱发多种染色体畸变,而且还能破坏精子线粒体功能,导致精子活力受损,受精率和受精指数下降,但其机制未明。本研究主要考察乙肝病毒S蛋白(HBs)对人精子氧化应激的影响。方法:人精子与不同浓度(0、25、50、100 μg/mL)HBs共孵育3 h后,用流式细胞术检测精子内活性氧(ROS)的变化情况,应用酶标仪检测精子细胞内丙二醛(MDA)含量和总抗氧化能力(TAC)。结果:人精子与25~100 μg/mL HBs共孵育3 h后,ROS阳性精子比率、人精子细胞膜MDA水平显著高于对照组,而TAC 水平显著低于对照组(P<0.05或0.01)。MDA水平以及ROS阳性精子比率的升高与HBs呈剂量依赖关系(r0.62,r0.75;P均<0.01)。TAC水平的降低与HBs呈剂量依赖关系(r-0.89;P<0.01)。结论:HBs可诱导人精子内ROS升高、TAC降低,从而产生氧化应激,使细胞膜脂质过氧化进而影响人精子的功能。

关键词: 乙肝病毒S蛋白, 氧化应激, 活性氧, 脂质过氧化

Abstract:

OBJECTIVE: It has been demonstrated that HBV is able not only to pass through the blood-testis barrier and integrate into sperm genome, leading to increase of the instability of sperm chromosomes, resulting in a various types of chromosomal aberrations, but also to destroy mitochondrial functions and induce loss of mitochondrial membrane potential (MMP), causing low sperm motility and reducing fertilization rate and fertilizing index. However, the exact pathogenic mechanism of such events is largely unknown so far. The purpose of this study is to explore the relationship between hepatitis B virus S protein (HBs) and oxidative stress in human sperm cells. METHODS:After the co-incubation of sperm cells with (0, 25, 50, 100 μg/mL) HBs, the reactive oxygen species (ROS) production was assessed by flow cytometric analyses;the total antioxidant capacity (TAC) level and the Aldetect (MDA-specific) lipid peroxidation (LP) level of the human sperm were measured by microplatereader. RESULTS:After incubation with 25 μg/mL of HBs for 3 h, the average rates of ROS-positive cells was significantly increased in the test groups as compared to those in the control groups, while TAC level was decreased when compared with the control. There was significantly higher level of MDA in the sperm cells exposed to 50 μg/mL of HBs for 3 h than that in the controls (P<0.05 or 0.01). HBs increased the MDA levels and the numbers of ROS-positive cells in a dose-dependent manner. HBs decreased the TAC levels in sperm cells in a dose-dependent manner. CONCLUSION:HBs exposure could lead to ROS generation, lipid peroxidation, TAC reduction, resulting in loss of sperm membrane integrity and causing sperm dysfunctions.

Key words: hepatitis B virus S protein, oxidative stress, reactive oxygen species, malondialdehyde

中图分类号:

R394.6

康祥锦, 丁悦, 杨洁, 杜红姿, 刘见桥, 黄天华. 乙肝病毒S蛋白对人精子氧化应激的影响[J]. 癌变·畸变·突变, 2015, 27(3): 202-206.

KANG Xiangjin, DING Yue, YANG Jie, DU Hongzi, LIU Jianqiao, HUANG Tianhua. Effects of hepatitis B virus S protein exposure on human sperm oxidative stress[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2015, 27(3): 202-206.

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乙肝病毒S蛋白对人精子氧化应激的影响

Effects of hepatitis B virus S protein exposure on human sperm oxidative stress

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