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目的探讨高温对雄性生殖细胞的氧化损伤作用及川芎嗪对细胞的保护作用。方法以猪睾丸细胞为研究对象,分为高温(43℃1 h、43℃2 h)组、高温+37℃6 h组、高温+川芎嗪(80、160和320μg/ml)组和阴性对照组,MTT比色法检测细胞损伤作用,采用激光共聚焦显微镜和流式细胞仪结合细胞活性氧检测试剂盒观察细胞内活性氧水平变化。结果在本实验条件下,高温对睾丸细胞没有明显的细胞损伤作用;43℃作用细胞1 h,细胞内活性氧未见明显升高(P>0.05),43℃作用细胞2 h,细胞内活性氧显著升高(P<0.05),即使脱离高温环境,短时间内活性氧水平亦未见降低;160μg/ml和320μg/ml川芎嗪能明显降低高温所致细胞内活性氧水平。结论高温能使睾丸细胞内活性氧生成增加,诱发氧化损伤,川芎嗪能抑制高温所致细胞内活性氧的升高,对细胞具有保护作用。
Objective To investigate the effect of high temperature on the oxidative damage of male germ cells and the protective effect of ligustrazine on the cells. Methods Pig testicular cells were divided into high temperature (43 ℃ 1 h, 43 ℃ 2 h) group, high temperature + 37 ℃ 6 h group, high temperature + ligustrazine (80, 160 and 320 μg / ml) group and negative control The changes of reactive oxygen species (ROS) in cells were observed by MTT colorimetric assay and laser scanning confocal microscopy (FCM) combined with flow cytometry and cell viability assay kit. Results Under the experimental conditions, high temperature had no obvious cell damage on testicular cells. At 43 ℃ for 1 h, there was no significant increase in intracellular reactive oxygen species (P> 0.05), at 43 ℃ for 2 h and intracellular activity Oxygen was significantly increased (P <0.05), and the level of reactive oxygen species did not decrease in a short period of time even after leaving high temperature environment. Ligustrazine at 160μg / ml and 320μg / ml significantly reduced intracellular reactive oxygen species. Conclusion High temperature can increase reactive oxygen species in testis cells and induce oxidative damage. Tetramethylpyrazine can inhibit the increase of intracellular reactive oxygen species (ROS) induced by high temperature and has a protective effect on cells.