用蛋白质组学技术探讨补肾生精法对老龄大鼠睾丸生精功能影响的机制  

Effect of Tonifying Kidney to Produce Sperm Therapy on Aged Rats’Spermatogenesis with Proteomics Technology

作　　者：陈通文[1] 曾金雄 王海鹰[1] 黄锦燕[1] 

机构地区：[1]广东省中医院珠海医院男科,广东珠海519015 [2]南京军区福州总院,福建福州350000

出　　处：《上海中医药大学学报》2008年第5期55-58,共4页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：广东省自然科学基金资助项目(06028730)

摘　　要：目的:应用蛋白质组学技术研究补肾生精法对老龄大鼠睾丸生精功能影响的机制。方法:20只22月龄老龄雄性SD大鼠随机分为2组,每组10只,分别为老龄组(SC)和补肾生精组(KT)。KT组灌胃给药补肾生精颗粒,连续用药60d。取附睾尾精子应用精子运动CASA分析技术,研究补肾生精法对衰老大鼠精子运动能力的影响。应用流式细胞仪检测技术,观察补肾生精颗粒对老龄大鼠睾丸生殖细胞凋亡率的影响。取大鼠右侧睾丸,应用双向凝胶电泳分析老龄组和补肾生精组大鼠睾丸表达差异蛋白。结果:与老龄组相比,补肾生精组大鼠精子密度明显增加(P<0.05);活动率和直线活动率显著上升(P<0.001);补肾生精组生殖细胞凋亡率明显下降(P<0.05)。与老龄组相比,补肾生精组大鼠睾丸中有7种蛋白表达发生了显著变化,其中4种蛋白表达上调,3种下调。对7种差异表达蛋白点胶内酶切后进行质谱分析,其中5种蛋白分别鉴定为磷脂酰乙醇胺结合蛋白(PEBP),热休克蛋白8(HSP7C),磷酸丙糖异构酶(Triose-phosphateisomerase,TPIS),3-磷酸甘油醛脱氢酶(GAPDH),谷胱苷肽转移酶1(GSTM1-1)。结论:补肾生精法对老龄大鼠睾丸功能具有提高精子运动能力及减少其生殖细胞凋亡的作用。运用蛋白质组学技术,研究补肾生精法对衰老过程睾丸蛋白表达的影响,有助于在蛋白质组水平阐释中医肾主生殖,延缓睾丸衰老,改善睾丸生精功能的机制。Objective： To study the mechanism of prompting spermatogenesis in aged rats with tonifying kidney to produce sperm therapy. Methods： The aged male SD rats of 22 months old were randomly devided into control group （SC）and tonifying kidney to produce sperm （KT） group. The rats of KT group were given ＂Bushen Shengjing Granule＂ by garage for 60 days. The epididymal sperms were tested with CASA system to investigate the influence of tonifying kidney to produce sperm therapy on the sperm motility of the aged rats. The flow cytometry assay was adopted to observe the the influence of ＂Bushen Shengjing Granule＂ on apoptosis rate of testicular germ cell in aged rats. The fight testises were detected with two-dimensional gel eleetrophoresis to analyze the differentially expressed proteins of the two groups. Results： Compared with control group, the sperm density of the KT group increased significantly（P〈 0.05）; both motility and linear motility of the treatment group rose significantly（P〈 0.001）; and the apoptosis rate of testicular germ cell in KT group decreased significantly （P〈 0.05）. Besides, there were significant changes of seven proteins＇ expression with four up-regulated and three down-regulated. Conclusion： The tonifying kidney to produce sperm therapy can improve sperm motility as well as reduce the apoptosis of germ cell in aged rats. Application of proteomies technology in research of tonifying kidney to produce sperm therapy＇s effect on expression of testieular protein in ageing process is conducive to explain the TCM theory of kidney dominating reproduction as well as the mechanism of the therapy＇ effect in resisting testicle aging and improving the spermatogenesis.

关 键 词：补肾生精 老龄大鼠 睾丸 精子功能 细胞凋亡 蛋白表达谱 

分 类 号：R256.5[医药卫生—中医内科学]