微波辐射致睾丸早期损伤的蛋白质组学研究

目的 探讨微波辐射早期睾丸蛋白质表达谱的改变及其意义.方法 24只雄性Wistar大鼠随机分为对照组(n=6)和辐射组(n=18).辐射组接受30mW/cm~2微波辐射,并分别于辐射后6、24、72h处死6只大鼠;对照组不接受辐射,于72h时间点处死.观察两组大鼠睾丸组织的病理学改变;采用双向凝胶电泳(2-DE)-基质辅助激光解吸电离飞行时间质谱(MAIDI-TOF-MS)-生物信息学分析比较蛋白质组学技术检测辐射72h后睾丸组织蛋白质表达谱的改变.结果 微波辐射后72h内,辐射组大鼠睾丸出现不同程度的损伤性改变,主要表现为生精细胞变性、坏死和脱落,精子减少或缺失,生精小管腔内红细胞和血浆蛋白积聚,间质水肿,血管淤血.对照组、辐射组(辐射后72h)睾丸组织蛋白质点分别为1 379±166、1 509±243个,匹配的蛋白质点为1 178±102、1 217±135个,匹配率为85.4%、80.6%.对两组进行匹配分析,发现差异表达的蛋白质点共136个,其中28个在辐射组上调,108个在辐射组下调.对其中的26个差异蛋白质点的鉴定结果显示,蛋白功能涉及细胞骨架、能量代谢、分子伴侣、离子结合运输、应激、信号转导等.结论 微波辐射可导致睾丸组织一系列蛋白表达发生改变,这些蛋白可能参与了生精细胞损伤的病理生理过程.

Proteomics analysis of the early testicular injury induced by microwave radiation

Objective To explore the changes in the early proteome maps of testis injured by microwave radiation,and evaluate their significance Methods Twenty-four male Wistar rats were randomly divided into control group (n=6) and radiation group (n=18).Rats in radiation group were sacrificed respectively at 6,24 and 72h after 30mW/cm~2 microwave radiation (6 each),and the animals in control group received no radiation. The pathological changes in testis tissue in the two groups were observed by light microscopy,and the comparative proteomie approach. based on two dimensional gel electrophoresis(2-DE),matrix-assisted laser desorption/ionisatiom-time of flight mass spectrometry (MALI-TOF-MS) identification and bioinformatics analysis,was employed to study the changes in the testis proteome maps at 72h after radiation.Results Degeneration,necrosis and shedding of the seminiferous epithelial cells,decrease or loss of spermatozoa,and accumulation of red blood cells and plasma protein in the lumen of the seminiferous tubule were observed,and interstitial edema and congestion of blood vessel were also found within 72h after radiation.There were 1379±166 and 1509±243 protein spots in the control group and radiation group at 72h after radiation respectively,the average matched spots were 1178±102 and 1217±135,and the average matching rate was 85.4%and 80.6%respectively.One hundred and thirty-six differential protein spots were found by matching analysis between the two groups,among which 28 spots were up-regulated and 108 were down-regulated in the radiation group. Identification of 26 differential expressed protein spots showed that the protein functions involved cytoskeleton,energy metabolism, molecular chaperones,ion binding and transportation,stress and signal transduction,and so on.Conclusion Microwave radiation may induce changes of series protein expression of the testis tissue,which may participate in the patho-physiological process of spermatogenic cell injury.