摘要
目的探索在小鼠月经发生的关键时期前回植孕酮对内质网应激诱导的自噬相关信号通路PERK/eIF2α/ATF4及凋亡关键信号分子CHOP和Caspase12的影响。方法 30只健康成年雌性 C57BL/6J 小鼠在建立生理性孕酮撤退月经模型的基础上,在孕酮撤退后12 h(12 h组)、16 h(16 h组)及孕酮撤退12 h回植孕酮后4 h(孕酮回植组)(每个时间点10只小鼠)处死小鼠并收获双侧子宫组织,通过蛋白印迹法和免疫组化法检测t-PERK、p-PERK、t-eIF2α、p-eIF2α、ATF4、CHOP、Caspase12蛋白在小鼠子宫内膜组织中的表达和定位,real-time PCR检测上述信号分子mRNA的表达。结果小鼠全子宫组织中t-PERK和t-eIF2α蛋白的表达量在各组间无显著差异( P >0.05),而PERK、eIF2α mRNA和p-PERK、p-eIF2α蛋白的表达量在12 h组和回植组未见显著差异,但都显著高于16 h组( P <0.05)。ATF4 mRNA和蛋白的表达趋势与PERK、eIF2α mRNA和p-PERK、p-eIF2α蛋白的表达趋势一致。p-PERK、p-eIF2α、ATF4蛋白主要定位在基底层与蜕膜化基质细胞交界处的基质细胞细胞质。CHOP、Caspase12 mRNA及蛋白的表达量在12 h组和回植组之间未见显著差异性表达,且都显著低于16 h组( P <0.05)。其蛋白主要定位在充分蜕膜化的基质细胞及腺上皮的细胞质。结论在月经发生关键时期前(12 h)回植孕酮,可以有效维持PERK/eIF2α/ATF4信号的表达并阻断凋亡信号通路成员CHOP、Caspase12表达,从而有效阻断月经发生。
Objective To investigate the effect of progesterone replantation before the critical period of menstruation on expression of endoplasmic reticulum stress-induced autophagy signaling pathway members,PERK/eIF2α/ATF4,and apoptosis key signaling molecules CHOP and Caspase12 in endometrial stromal cells of mice. Methods On the basis of the establishment of physiological progesterone withdrawal menstrual model,30 healthy adult female C57BL/6J mice were divided into 3 groups,12 h,16 h progesterone withdrawal groups and progesterone replantation group.Mice were sacrificed and bilateral uterine tissues were harvested at 12 h,16 h after progesterone withdrawal.In progesterone replantation group,progesterone was replanted at 12 h after progesterone withdrawal and executed 4 hours later.The expression and localization of t-PERK,p-PERK,t-eIF2α,p-eIF2α,ATF4,CHOP and Caspase12 proteins in mouse endometrial tissues were detected by Western blot and immunohistochemistry,the mRNA expression was detected by real-time PCR. Results There was no significant difference in the expression of t-PERK and t-eIF2α proteins in mouse uterine tissue among groups ( P >0.05).The expression of PERK,eIF2α mRNA and p-PERK,p-eIF2α proteins were not significantly different between 12 h group and the progesterone replantation group.But both groups were significantly higher than 16 h group ( P <0.05).The expression trends of ATF4 mRNA and protein was consistent with that of PERK,eIF2α mRNA and p-PERK,p-eIF2α proteins.The proteins of p-PERK,p-eIF2α and ATF4 are mainly exepressed in the cytoplasm of decidual stromal cells which located at the junction of basal layer and decidualized stromal cells.The expression levels of mRNA and proteins of CHOP and Caspase12 were not significantly different between the 12 h group and the progesterone replantation group,but both groups were significantly lower than 16 h group ( P <0.05).The proteins of CHOP and Caspase12 are mainly localized in cytoplasm of adequately decidualized stromal cells and the glandular
作者
于明明
谢卓
金华
王乾兴
Yu Mingming;Xie Zhuo;Jin Hua;Wang Qianxing(Department of Cell Biology,Zunyi Medical University,Zunyi Guizhou 563099,China;Department of Medical,Xishui People's Hospital,Zunyi Guizhou 564600,China)
出处
《遵义医学院学报》
2019年第3期276-281,共6页
Journal of Zunyi Medical University
基金
贵州省科技厅联合基金资助项目(NO:黔科合J字LKZ[2010]33)
遵义市科学技术基金资助项目(NO:遵市科合社字[2018]24,[2018] 266)
关键词
内质网应激
自噬
凋亡
孕酮撤退
月经
ERS
autophagy
apoptosis
progesterone withdrawal
menstruation
