葛根素对热应激致牛睾丸支持细胞氧化损伤的保护作用

Protective Effect of Puerarin on Heat Stress-induced Oxidative Damage in Bovine Sertoli Cells

葛根素(Pue)是从中药野葛或甘葛藤根中提取的黄酮类化合物,有明显的抗氧化功能。Pue是否有对热应激时牛睾丸支持细胞(SCs)保护作用尚不清楚。本研究采用42℃热应激1 h,然后34℃恢复6 h。用CCK-8检测细胞存活率;用Hoechst33258检测细胞凋亡;用分光光度法检测细胞丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和过氧化氢酶(CAT)的变化,以及Western-Blot检测HSP70蛋白表达。结果显示,热应激后细胞存活率下降,细胞发生凋亡,产生氧化损伤。葛根素和维生素E可以极显著降低热应激时细胞MDA含量,显著增加SOD、CAT和GSH-Px活力。Pue与维生素E组相比无显著性差异。此外,葛根素可显著增加受热应激SCs中HSP70蛋白的表达。上述结果表明,葛根素能减少热应激下SCs的氧化损伤,其机制可能与提高抗氧化酶活性及增加热休克蛋白HSP70的表达有关。

Puerarin（Pue） is a flavonoid extracted from traditional Chinese medicine Puerarin with obvious antioxidant effect.Whether Pue has the protective effect of Bovine Sertoil cells（SCs） on heat stress is still unclear.The cells were incubated with or without various concentrations of Pue,and then heat-stressed at 42℃ for 1 h and returned to 34℃ for 6h.Cell viability was decreased,and the apoptosis rate was increased and the cellular oxidative damage occurred after heat stress.However,the MDA content in the SCs treated with heat stress decreased significantly after Pue pretreatment.Also,the SOD,CAT and GSH-Px activities were significantly increased compared to heat stress group.There was no significant difference in the parameters determined above between Pue and VE pretreatment groups.Furthermore,Western-blot results showed that Pue significantly enhanced the expression of HSP70.Pue protects SCs from heat stress induced oxidative damage possibly by increasing activities of antioxidative enzymes and increased HSP70 expression.