摘要
目的探讨不同表达水平的钙网蛋白(calreticulin,CRT)与先兆流产的关系。方法 (1)分别应用免疫组织化学法、Western blotting和q PCR检测早期正常妊娠(n=35,均因非意愿妊娠而要求人工流产)及早期先兆流产患者人工流产后的蜕膜和绒毛组织中CRT蛋白及mRNA的表达。(2)分别转染合成并经检测的质粒m Emerald-CRT与CRT-siRNA至BeWo细胞中,检测CRT及Caspase-3的表达。结果免疫组织化学可见CRT阳性信号定位于绒毛两层滋养层细胞的胞质和胞膜上,先兆流产组染色比正常早孕组深、面积大;q PCR与Western blotting检测CRT表达量在先兆流产组明显高于正常早孕组,先兆流产组中mRNA表达量为正常早孕组的1.2倍(P<0.05)。q PCR与Western blotting检测CRT与Caspase-3表达量在质粒(m Emerald-CRT)转染组高于对照组,质粒转染组中CRT及Caspase-3的表达量分别是对照组的3.5倍与2倍(P<0.05),在si-RNA组低于对照组,对照组中CRT及Caspase-3的表达量分别si-RNA组的5倍与2倍。结论早期先兆流产的发生可能与CRT在绒毛和蜕膜组织中的高表达并介导凋亡相关。
Objective To explore the relationship between the expression level of calreticulin （CRT） and early threatened abortion by observing CRT expression level in decidua and villi tissues. Methods 1） The decidua and villi tissues of normal pregnancies and early threatened abortion patients were collected and treated respectively. 2） The expression of CRT protein was verified by immunohistochemistry and Western blotting, and the expression of CRT mRNA was detected by qPCR. 3） The plasmid mEmerald-CRT was synthesized and detected. 4） CRT-siRNA was synthesized and detected. 5） Plasmid mEmerald-CRT and CRT-siRNA were transfected into BeWo cells respectively and the expressions of CRT and Caspase-3 were detected by Western blotting and qPCR. Results 1） The decidua and villi tissues of normal pregnancies （35 cases） and early threatened abortion （40 cases） were collected. 2） Immunohistochemistry showed that the stains were deeper and bigger in early threatened abortion group than the normal pregnancy group; qPCR test and Western blotting showed the expression of CRT was significantly higher in early threatened abortion than that in normal pregnancy, the expression ofmRNA in early threatened abortion groups was 1.2 times of normal pregnancy （P〈0.05）. 3） qPCR test and Western blotting showed CRT and Caspase-3 in mEmerald-CRT groups were significantly higher than those in control groups, the expressions of CRT and Caspase-3 in mEmerald-CRT groups were 3.5 times and 2 times of control groups respectively （P〈0.05）. 4） qPCR test and Western blotting showed CRT and Caspase-3 in CRT-siRNA groups were significantly lower than those in control groups, the expressions of CRT and Caspase-3 in CRT-siRNA groups were 5 times and 2 times of control groups, respectively （P〈0.05）. Conclusion The occurrence of early threatened abortion may be related to excessive CRT expression and its mediated abnormal apoptosis in decidua and villi tissues.
作者
邓小月
柳长柏
熊焱强
秦琪
李志英
Deng Xiaoyue;Liu Changbai;Xiong Yanqiang;Qin Qi;Li Zhiying(The Second Medical School, China Three Gorges University, Yichang 443001, China;Three Gorges University Hubei Key Laboratory of Tumor Microenvironment and Immunotherapy, Yichang 443002, China)
出处
《中华生殖与避孕杂志》
CAS
CSCD
北大核心
2017年第11期888-895,共8页
Chinese Journal of Reproduction and Contraception
关键词
钙网蛋白
绒毛
蜕膜
早期先兆流产
细胞凋亡
Calreticulin
Decidua
Villi
Early threatened abortion
Cell apoptosis
