中国农业科学 ›› 2009, Vol. 42 ›› Issue (11): 4064-4068 .doi: 10.3864/j.issn.0578-1752.2009.11.038

• 畜牧·资源昆虫 • 上一篇    下一篇

热应激对猪睾丸细胞活性氧水平影响

张晓峰,刘迪,王淼,李垚

  

  1. (东北农业大学动物科学技术学院)
  • 收稿日期:2009-04-30 修回日期:2009-07-02 出版日期:2009-11-10 发布日期:2009-11-10
  • 通讯作者: 李垚

Effects of Heat Stress on Reactive Oxygen Species in Swine Testicular Cells

ZHANG Xiao-feng, LIU Di, WANG Miao, LI Yao   

  1. (东北农业大学动物科学技术学院)
  • Received:2009-04-30 Revised:2009-07-02 Online:2009-11-10 Published:2009-11-10
  • Contact: LI Yao

摘要:

【目的】探讨热应激对猪睾丸细胞的氧化损伤。【方法】将猪睾丸细胞分为43℃ 1 h组、43℃ 2 h组、43℃ 1 h+37℃ 6 h组、43℃ 2 h+37℃ 6 h组、正常对照组及阳性对照组,采用光镜和电镜观察细胞及线粒体形态学变化,采用细胞活性氧检测试剂盒,通过激光共聚焦显微镜和流式细胞仪观察细胞内活性氧水平的变化。【结果】43℃处理1 h后,细胞变圆,线粒体发生部分或广泛变性,43℃ 2 h组和43℃ 2 h+37℃ 6 h组更为明显;43℃ 1 h组细胞内活性氧水平未见明显升高(P>0.05),但43℃ 2 h组细胞内活性氧水平显著高于43℃ 1 h组(P<0.05)。上述2组细胞经43℃ 处理后37℃继续孵育6 h,细胞内活性氧水平均明显增强(P<0.05)。【结论】热应激使睾丸细胞线粒体形态学发生改变,诱发细胞氧化损伤,进而可能影响睾丸的生殖功能。

关键词: 热应激, 睾丸细胞, 活性氧, 线粒体

Abstract:

【Objective】 To study the effect of heat stress on oxidative injuries in swine testicular cells. 【Method】 Swine testicular cells were divided into 6 groups, 43℃ 1 h, 43℃ 2 h, 43℃ 1 h+37℃ 6 h, 43℃ 2 h+37℃ 6 h, negative and positive groups. Cellular and mitochondrial morphology were observed by using light microscope and transmission electron microscope. Laser confocal microscopy and flow cytometry were used to determine the level of reactive oxygen species (ROS) in testicular cells with ROS test Kit. 【Result】 After 1 h with 43℃ treatment, cells became circular and mitochondria showed partial or extensive denaturation, especially obvious in 43℃ 2 h and 43℃ 2 h+37℃ 6 h groups. There were no significant increase of ROS in testicular cells in 43℃ 1 h group (P>0.05), but the level of ROS obviously increased in 43℃ 2 h group, compared to the former (P<0.05), and the level of ROS in 43℃ 1 h and 43℃ 2 h groups also continued to increase after additional 6 h in 37℃, compared to negative group (P<0.05). 【Conclusion】 Heat stress could result in the morphological changes of mitochondria of testicular cells and induce oxidative injuries, thus could affect the reproductive function of testis.

Key words: heat stress, testicular cells, reactive oxygen species, mitochondria