中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (18): 3361-3364.doi: 10.3969/j.issn.1673-8225.2011.18.032

• 器官移植基础实验 basic experiments of organ transplantation • 上一篇    下一篇

氧自由基对生殖细胞三磷酸腺苷酶和生长抑素的影响

胡建新1,何  坚1,刘  军1,孙兆林1,王元林1,刘修恒2   

  1. 1贵州省人民医院泌尿外科,贵州省贵阳市 550002
    2武汉大学人民医院泌尿外科,湖北省武汉市  430064
  • 收稿日期:2010-11-19 修回日期:2011-02-14 出版日期:2011-04-30 发布日期:2011-04-30
  • 通讯作者: 何坚,主任医师,硕士生导师,贵州省人民医院泌尿外科,贵州省贵阳市 550002 gzhejian@ yahoo.com.cn
  • 作者简介:胡建新★,男,1970年生,湖北省咸丰县人,土家族,2006年贵阳医学院毕业,硕士,副主任医师,主要从事泌尿外科及男科研究。 hjx918@163. com
  • 基金资助:

    贵州省卫生厅科技基金(gzwkj2009-1-055),课题名称:自由基对生殖细胞ATPase和生长抑素影响的实验与临床研究。

Effect of oxygen free radicals on adenosine triphosphatase and somatostatin in reproductive cells

Hu Jian-xin1, He Jian1, Liu Jun1, Sun Zhao-lin1, Wang Yuan-lin1, Liu Xiu-heng2   

  1. 1Department of Urology, Guizhou Provincial People’s Hospital, Guiyang  550002, Guizhou Province, China
    2Department of Urology, Renmin Hospital of Wuhan University, Wuhan  430064, Hubei Province, China
  • Received:2010-11-19 Revised:2011-02-14 Online:2011-04-30 Published:2011-04-30
  • Contact: He Jian, Chief physician, Master’s supervisor, Department of Urology, Guizhou Provincial People’s Hospital, Guiyang 550002, Guizhou Province, China gzhejian@yahoo. com.cn
  • About author:Hu Jian-xin★, Master, Associate chief physician, Department of Urology, Guizhou Provincial People’s Hospital, Guiyang 550002, Guizhou Province, China hjx918@163.com
  • Supported by:

    the Science and Technology Foundation of Guizhou Health Committee, No. gzwkj2009-1-055*

摘要:

背景:有关细胞自由基的研究,主要集中在动物体内细胞自由基的产生与清除方面。
目的:观察氧自由基对体外培养的睾丸生殖细胞三磷酸腺苷酶(ATPase)和生长抑素含量的影响。
方法:体外分离、纯化小鼠睾丸生殖细胞,培养24 h后,将其分为2组,实验组添加黄嘌呤-黄嘌呤氧化酶建立氧自由基损伤模型;对照添加生理盐水。继续培养24,48 h,采用酶组化及免疫组化染色观察各组ATPase和生长抑素表达变化。采用LUZEX-F图像分析仪分别测定其灰度值;用酶联免疫检测仪测定492 nm波长的吸光度值。
结果与结论:给药后24,48 h,实验组ATPase、生长抑素灰度值显著高于对照组(P < 0.01),吸光度值显著低于对照组( < 0.01)。提示氧自由基可降低ATPase和生长抑素含量,影响生殖细胞在体外培养过程中的生长与增殖。

关键词: 睾丸生殖细胞, 体外培养, 氧自由基, 三磷酸腺苷酶, 生长抑素

Abstract:

BACKGROUND: Most of studies about oxygen free radicals focus on production and scavenging of free radicals in animal cells.
OBJECTIVE: To investigate the effect of oxygen free radical on adenosine triphosphatase (ATPase) and somatostatin (SOM) in spermatogonial cells cultured in vitro.
METHODS: Spermatogonial cells isolated from mice were purified and cultured in vitro and divided into two groups after 24-hour culture. In the experimental group, xanthine-xanthinoxidase (X-XO) was used to establish an oxygen free radical damaging model. In the control group, normal saline was added. After 24 and 48 hours, changes of ATPase and SOM were observed in the two groups. Gray value and absorbance value at 492 nm were determined.
RESULTS AND CONCLUSION: After 24 and 48 hours, the gray values of ATPase and SOM in the experimental group were significantly higher than those in the control group (P < 0.01), while the absorbance values in the experimental group were lower than those in the control group (P < 0.01). These indicated that oxygen free radicals can decrease the ATPase and SOM, and influence the growth and proliferation of spermatogonial cells cultured in vitro.

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