为了探讨温和热应激、脂多糖(LPS)和谷氨酰胺(Gln)诱导牛睾丸支持细胞(b SCs)HSP72的时效表达规律,本试验将传至第3代b SCs分成热应激、LPS和Gln处理组,用CCK-8试剂盒检测细胞活性,用RT-PCR和Western-Blot方法分别检测处理后0、2、4、6、8、12、14h和16 h时细胞内HSP72 mRNA和蛋白表达。结果显示,3种方式均能诱导HSP72表达,且HSP72蛋白分别在热应激、Gln和LPS预处理后2、4h和12 h时表达量最高。结果表明,热应激或Gln均比LPS诱导牛睾丸支持细胞HSP72表达的时间早,提示可考虑用Gln减少由LPS诱导的细胞损伤。
This study was aimed to investigate HSP72 aging expression pattern of bovine Sertoli cell(bSCs) induced by pretreatments with mild heat stress,LPS and Gin. The bSCs on their third generation were used and divided into heat stress, LPS and Gin stimulating groups. The cell viability was detected by CCK - 8. The cellular HSP72 mRNA and its protein expression levels were detected at 0,2,4,6,8,10,12,14 and 16 h after treatments by RT - PCR and Western - Blot, respectively, The results showed that three measures of pretreatment could all induce HSP72 expression, and the highest protein expression levels were appeared at 2,4 and 12 h after heat stress,Gin and LPS pretreatment, respectively. From the above results we concluded that heat stress and Gin pre - stimulation can induce HSP72 expression in advance in bovine Sertoli cells,which indicate that Gin may be used for ameliorates LPS induced cellular injury.