梗阻性无精子症患者睾丸精子左旋肉碱培养后关键基因表达量的变化研究  

Expressions of sperm-specific genes in carnitine-cultured testis sperm of obstructive azoospermia patients

作　　者：师娟子 张姗姗[2] 张洲 梁倩[2] 石玉[2] 华进联[2] 孙婷 

机构地区：[1]陕西省妇幼保健院辅助生殖中心,陕西西安710003 [2]西北农林科技大学动物医学院,陕西省干细胞工程技术研究中心,陕西省农业分子生物学重点实验室,陕西杨凌712100

出　　处：《中华男科学杂志》2010年第6期504-509,共6页National Journal of Andrology

基　　金：陕西省科技厅科技攻关资助[2007K14-02(1)];教育部重点科研项目(109148);中国博士后科学基金特别资助(200801438)课题~~

摘　　要：目的:探讨肉碱对人精子活力的影响及其对男性不育的治疗作用。方法:以梗阻性无精子症患者睾丸穿刺取得的精子为研究对象,通过普通培养液、培养液中添加100mmol/L及250mmol/L肉碱进行培养,比较培养前后精子活动率的变化,以及用RT-PCR检测男性生殖细胞特异性基因Vasa、Dazl、Acr、Prm1及ATPase6.0表达量的变化,探讨左旋肉碱与有关精子发生和成熟过程的重要功能基因表达的关系。结果:培养24～72h,添加100mmol/L的左旋肉碱培养液运动精子数明显较未添加和添加250mmol/L组高(P<0.01);100mmol/L左旋肉碱培养液组中典型的快速前向运动精子数明显较多,巴氏染色法观察其具有正常的精子结构,RT-PCR检测表明100mmol/L的肉碱可提高精子中Acr、Prm1、Dazl及ATPase6.0基因的表达量,而250mmol/L的肉碱组Dazl、Acr、Prm1基因的表达减少。结论:合适浓度的左旋肉碱可通过上调一些生殖相关基因的表达使得睾丸精子培养后活力增强,有利于对睾丸穿刺患者行单精子注射时精子的选择。如培养液中肉碱浓度过高,可能由于肉碱的毒性作用,反而使得这些基因的表达量减少。Objective： To investigate the effects of carnitine on human sperm motility and its potential role in the treatment of male infertility diseases. Methods ： We obtained sperm by testis puncture from obstructive azoospermia patients and cultured them in vitro with normal culture solution （the control group） and the solution with L-carnitine at the concentration of 100 and 250 mmol/L, respectively. We observed the changes in sperm motility and morphology before and after the treatment, detected the expressions of the germ-specific genes, Vasa, Dazl, Acr, Prm1 and ATPase6.0 by RT-PCR, and investigated the relationship between L-caruitine and the genes associated with sperm development and maturation. Results ： After 24 - 72 hours of treatment, the percentage of motile sperm was significantly higher in the 100 mmol/L L-carnitine group than in the control and 250 mmol/L L-carnitine groups （P 〈 0.01 ） ; the number of forward moving sperm was obviously increased and sperm morphology remained normal in the 100 mmol/L L-carnitine group.RT-PCR showed that L-carnitine increased the expressions of Acr, Prm1, Dazl and ATPase6. 0 at the concentration of 100 mmol/L, and decreased the expressions of Dazl, Acr and Prm1 at 250 mmol/L. Conclusion ： L-carnitine at a proper concentration may improve the motility of incubated testicular sperm by upregulating the expressions of some sperm-specific genes, which helps sperm selection for intracytoplasmic sperm injection. However, a higher concentration of L-carnitine may reduce the expressions of these genes, probably due to its increased toxicity.

关 键 词：左旋肉碱 精子活力 男性不育 RT-PCR 梗阻性无精子症 

分 类 号：R698.2[医药卫生—泌尿科学]