摘要
目的我们新近的研究表明,超生理剂量的皮质酮(大鼠体内的糖皮质激素)可通过激活caspase-3诱导大鼠Leydig细胞凋亡。本研究拟评价在皮质酮诱导的大鼠Leydig细胞凋亡过程中,caspase-3的激活是否有其上游的caspase-8和caspase-9的参与。方法采用荧光分光光度法检测经皮质酮处理的大鼠Leydig细胞中caspase-8活性,以DNA梯状电泳条带作为评价细胞凋亡的指标,观察caspase-8抑制剂是否能够抑制细胞凋亡,采用RT-PCR检测皮质酮诱导的大鼠Leydig细胞中caspase-9的mRNA水平。结果在皮质酮诱导的大鼠Leydig细胞中出现caspase-8活性增高,以12h最为显著,升高的caspase-8的活性可被caspase-8抑制剂抑制,并导致Leydig细胞的凋亡过程被阻断。Leydig细胞中caspase-9的mRNA水平在皮质酮作用下上升,同样以12h最为显著。结论皮质酮诱导的大鼠Leydig细胞凋亡与caspase-8和caspase-9有关。
Objective To observe whether the caspase-8 and caspase-9 are involved in the CORT-mediated Leydig cell apoptosis, Methods The caspase-8 activity in Leydig cell subjected to CORT treatment was detected by fluorescent quantitative assay. Apoptosis was demonstrated by DNA laddering electrophoresis. The levels of caspase- 9 mRNA expression in the apoptotic pathway were investigated by RT-PCR, Results Caspase-8 activity in Leydig cells increased at 6h after the start of CORT administration, peaked at 12h and declined thereafter, In the presence of a specific caspase inhibitor, IETD-FMK, Leydig cell apoptosis was suppressed and agarose gel electrophoresis showed that marked DNA laddering typical for apoptosis was not detectable in Leydig cells treated with a dose of 100nM CORT, The expression of caspase-9 mRNA is increased during apoptosis, Conclusion caspase-8 and caspase-9 are all implicated in the process of apoptotic Leydig cell induced by CORT,
出处
《中国男科学杂志》
CAS
CSCD
2005年第5期9-12,19,共5页
Chinese Journal of Andrology
