热休克蛋白90与人精子氧化应激相关性的初步研究

Preliminary Study on the Association between Heat Shock Protein 90 and Oxidative Stress in Human Spermatozoa

目的探讨热休克蛋白90(HSP90)与人精子氧化应激是否相关。方法人精子标本经percoll分离处理后,分别用含0、100、200和400μmol/L H2O2的获能培养基应激刺激2h,用免疫印迹法检测和灰度分析法定量分析了不同氧化应激状态下精子HSP90表达水平,用相同方法进一步考察200μmol/L H2O2应激刺激0、0.5、1和2h后HSP90表达水平。结果不同浓度H2O2应激刺激人精子2h后,各组间HSP90表达水平具有统计学差异,400μmol/L组HSP90表达量较0μmol/L组明显降低;200μmol/L H2O2应激刺激人精子不同时间后各组间HSP90表达水平亦具有统计学差异,在0.5h时HSP90表达水平最低,后随时间延长表达水平又逐步升高。结论氧化应激时人精子HSP90表达水平可发生变化,且与活性氧浓度和应激时间相关,推测HSP90可能参与人精子氧化应激过程。

Objective To investigate whether heat shock protein 90（HSP90） is involved in oxidative stress in human spermatozoa.Methods Spermatozoa were stimulated respectively by 0,100,200 and 400μmol/L H2 O2 for 2h in culture medium after the samples were separated and prepared by percoll.Western blot and gray intensity analysis were carried out to determine the levels of HSP90 expression.Furthermore,the levels of HSP90 expression were evaluated by the same method after spermatozoa were stimulated by 200μ,mol/L H2O2for 0,0.5,1 and 2h respectively.Results HSP90 expression levels had significant difference among groups treated with different concentrations of H2O2 for 2h.The level of 400μmol/L group was significantly lower than 0μmol/L group.HSP90 expression levels also appeared significantly different among groups which were stimulated for different lengths of times at H2O2 concentration of 200μmol/L.The level was lowest at 0.5h,then increased with the extended time of incubation.Conclusion Expression levels of HSP90 protein changed in response to oxidative stress and were associated with the concentration of ROS and the time of stimulation by oxidative stressor.It implys that HSP90 is involved in oxidative stress in human spermatozoa.