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  • 张衡,武琳琳,李世杰,匡野,马兴红.混合谱系激酶结构域样蛋白在小鼠妊娠早期子宫及蜕膜中的表达及调控[J].第二军医大学学报,2018,39(10):1122-1127    [点击复制]
  • ZHANG Heng,WU Lin-lin,LI Shi-jie,KUANG Ye,MA Xing-hong.Expression and regulation of mixed lineage kinase domain-like protein in uterus during early pregnancy and decidua in mice[J].Acad J Sec Mil Med Univ,2018,39(10):1122-1127   [点击复制]
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混合谱系激酶结构域样蛋白在小鼠妊娠早期子宫及蜕膜中的表达及调控
张衡1,武琳琳1,李世杰1,匡野2,马兴红1*
0
(1. 东北农业大学生命科学学院, 哈尔滨 150030;
2. 哈尔滨医科大学附属第二医院妇产科, 哈尔滨 150001
*通信作者)
摘要:
目的 探讨混合谱系激酶结构域样蛋白(MLKL)在小鼠妊娠早期子宫及蜕膜组织中的表达规律。方法 构建小鼠早期妊娠模型、人工诱导子宫蜕膜化模型、雌二醇和(或)孕酮处理子宫模型;体外培养人子宫内膜基质细胞,应用雌二醇、醋酸甲羟孕酮和二丁酰环腺苷酸诱导其蜕膜化。通过实时荧光定量PCR、原位杂交和蛋白质印迹法分析小鼠早期妊娠子宫、蜕膜化子宫和激素处理子宫中MLKL mRNA和蛋白的表达规律,通过实时荧光定量PCR检测MLKL mRNA在体外诱导的人蜕膜化细胞中的表达。结果 (1)在小鼠早期妊娠子宫中,MLKL mRNA及蛋白在妊娠第1~4天的子宫腔上皮表达量较低且不规律,在妊娠第5天的子宫腔上皮及其周围的蜕膜细胞中表达,妊娠第6~8天主要集中在蜕膜组织中表达,着床后表达量逐日增高并在妊娠第7天达到最高峰,妊娠第8天表达稍有下降。(2)在人工诱导蜕膜化的小鼠子宫中,MLKL mRNA表达量很高,整个蜕膜区域均有表达,而在对照组小鼠子宫中则无明显表达。MLKL蛋白的表达与mRNA相一致。MLKL mRNA在体外诱导的人蜕膜化细胞中的表达量高于对照组,差异有统计学意义(P<0.05)。(3)在孕酮处理的小鼠子宫中MLKL mRNA及蛋白表达量增高,与对照组相比差异有统计学意义(P<0.05)。结论 MLKL受孕酮的调控参与哺乳动物胚胎着床和蜕膜化过程。
关键词:  混合谱系激酶结构域样蛋白  程序性细胞坏死  妊娠  胚胎植入  蜕膜
DOI:10.16781/j.0258-879x.2018.10.1122
投稿时间:2018-06-07修订日期:2018-08-02
基金项目:国家自然科学基金(31271601,31571553),黑龙江省教育厅海外学人科研资助项目(1253HQ016).
Expression and regulation of mixed lineage kinase domain-like protein in uterus during early pregnancy and decidua in mice
ZHANG Heng1,WU Lin-lin1,LI Shi-jie1,KUANG Ye2,MA Xing-hong1*
(1. College of Life Science, Northeast Agricultural University, Harbin 150030, Heilongjiang, China;
2. Department of Obstetrics and Gynecology, the 2nd Affiliated Hospital of Harbin Medical University, Harbin 150001, Heilongjiang, China
*Corresponding author)
Abstract:
Objective To investigate the expression of mixed lineage kinase domain-like protein (MLKL) in uterus during early pregnancy and decidua in mice. Methods Different mouse models including early pregnancy model, artificially induced decidualization model and hormone and/or progesterone treatment of uterine model were constructed; human endometrial stromal cells were cultured in vitro and were induced for decidualization by treating with estradiol-17β, medroxyprogesterone acetate and dibutyryl cyclic adenosine monophosphate. The expression of MLKL mRNA and protein in the uterus of early pregnancy, decidual uterus, and hormone-treated uterus in mice were analyzed by real-time fluorescent quantitative PCR, in situ hybridization and Western blotting. The expression of MLKL mRNA in human decidual cells induced in vitro was detected by real-time fluorescent quantitative PCR. Results (1) In the uterus during early pregnancy in mice, the expression of MLKL mRNA and protein in uterine epithelium on the 1st to 4th day of pregnancy (day 1 was day of vaginal sperm) was low and irregular. It was expressed in the uterine epithelium and surrounding decidual cells on the 5th day of pregnacy, and was mainly expressed in the decidua from the 6th to 8th day of pregnancy. After the implantation, the expression of MLKL mRNA and protein was day-by-day increased and reached the highest on the 7th day of pregnancy, with a slight decrease on the 8th day. (2) In the uterus of mice with artificially induced decidualization, MLKL mRNA was expressed in the entire decidual region with high level; while there was no significant expression in the uterus of the control mice. The expression of MLKL protein was consistent with the expression of MLKL mRNA. The expression of MLKL mRNA in human decidual cells induced in vitro was significantly higher than that in the control group (P<0.05). (3) The expression of MLKL mRNA and protein in progesterone-treated uterus was significantly increased compared with the control group (P<0.05). Conclusion MLKL regulated by steroid hormone progesterone is involved in embryo implantation and decidualization of mammals.
Key words:  mixed lineage kinase domain-like protein  necroptosis  pregnancy  embryo implantation  decidua