高温对原代支持细胞增殖能力和occludin紧密连接蛋白的影响(英文)

The Effect of Hyperthermia on Proliferation and Occludin Expression of Rat Sertoli Cells in Vitro

目的:研究高温对原代大鼠睾丸支持细胞增殖作用和对紧密连接蛋白的损伤机制。方法:体外分离培养雄性Wistar大鼠睾丸支持细胞(Sertoli cell,SC),免疫组化FasL鉴定。实验分为对照组(36℃)和实验组(37℃、38℃、39℃)。在相应的温度下培养5d后,CCK-8法检测SC增殖作用,Western印迹法检测OCLN表达水平。结果:体外培养SC的纯度>90%,CCK-8结果显示,细胞增殖活性在36℃时最强,37～39℃时逐渐降低;Western印迹显示,36℃时OCLN表达量最高;高于36℃时,OCLN表达量随着温度的增加而降低(P<0.05)。结论:高温影响SC增殖活性,破坏了支持细胞紧密连接蛋白的正常表达和血睾屏障的完整性,从而影响正常精子的形成。

Objective： To investigate the mechanism of injury caused by hyperthermia to rat sertoli cells（SCs） by detecting the cell proliferation and the expression of occludin of SCs in vitro. Methods： The SCs were isolated from the testicle of male Wistar rat, and cul- tured in vitro and identified by FasL. The SCs were divided into tour groups： control group （36℃） and experimental groups （37℃, 38℃, 39℃）. After exposed under the above temperatures for 5 days of in vitro culture, the cell proliferation was measured by CCK-8 assay, and the OCLN expression was detected by western blotting. Results： The result of FasL indicated that the purity of SCs was more than 90%. CCK-8 assay showed that the proliferative ability of SCs was file highest under 36℃, and decreased gradually when the temperature rises from 37℃ to 39℃. Meanwhile, the expression of OCLN was similar to the proliferative ability： the expression was the strongest at 36℃, and declined under 37℃, 38℃and 39℃（P〈0.01）. Conclusion： Higher temperatures （such as 37℃ ,38℃ and 39℃ in this experiment） may inhibit the proliferative ability of rat SCs, and also reduce the expression of OCLN of SCs, which may be one of the contributing aspects leading to the impairment of male soermato^enesis by high temoerature.