宫腔注射PBMCs改善小鼠胚泡着床障碍及其机制初探  

Improvement of Mouse Blastocyst Implantation by Intrauterine Injection of PBMCs and Its Mechanism

作　　者：余楠 李伟[1] 林星光[1] 陈素华[1] 冯玲[1] 邓东锐[1] 曾万江[1] 乌剑利[1] YU Nan;LI Wei;LIN Xing-guang;CHEN Su-hua;FENG Ling;DENG Dong-rui;ZENG Wan-jiang;WU Jian-li(Department of Obstetrics and Gynecology,Tongji Hospital of Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属同济医院产科,武汉430000

出　　处：《微循环学杂志》2020年第3期5-10,共6页Chinese Journal of Microcirculation

基　　金：国家自然科学基金资助项目(81701530,81701476);湖北省科技计划项目(2017CFB626)。

摘　　要：目的:探讨宫腔注射外周血单个核细胞(PBMCs)对着床窗期着床障碍模型小鼠胚泡着床的影响及其机制。方法:选取SPF级性成熟昆明雌性小鼠,将与同批雄性小鼠交配后发现阴栓者(妊娠第一天,Pd1)分为对照组、着床障碍组和PBMCs处理组,每组28只。PBMCs处理组于Pd2宫腔内注射PBMCs 2.5μl(1-2×10^7个/ml);着床障碍组和PBMCs处理组于Pd2早9∶00皮下注射米非司酮(0.08mg/0.1ml)复制着床障碍模型。对照组注射等量磷酸盐缓冲液(PBS)。Pd4晚10∶00处死各组部分小鼠,采用免疫组化染色法和Real Time-PCR检测着床窗期子宫内膜血管内皮生长因子(VEGF)及整合素β3蛋白和mRNA表达水平,Pd8处死各组其余小鼠,观察胎泡着床情况,计算各组着床率。结果:与对照组相比,着床障碍组小鼠的着床率和着床窗期VEGF及整合素β3蛋白和mRNA表达明显降低(P<0.01);PBMCs处理组小鼠着床率和着床窗期VEGF及整合素β3蛋白和mRNA表达较着床障碍组显著增加(P<0.01),且接近对照组水平(P>0.05)。结论:PBMCs宫腔内注射能提高着床障碍模型小鼠妊娠率,其机制可能与其增加着床窗期子宫内膜VEGF和整合素β3表达,从而改善小鼠子宫内膜容受性有关。Objective: To investigate the effect of PBMCs intrauterine injection on blastocyst implantation and its mechanism in blastocyst implantation dysfunctional(BID) mice. Method: After mating with male mice, Kunming female mice with SPF grade sexual maturity were divided into control group, BID group and PBMCs treatment group on the first day of pregnancy(Pd1). PBMCs 2.5μl(1-2×10~7/ml) were injected into the uterine cavity of Pd2 in PBMCs treatment group, and mifepristone(0.08 mg/0.1 ml) was injected subcutaneously into the BID group and PBMCs treatment group at 9∶00 AM of Pd2. The control group was injected with the same amount of phosphate buffer(PBS). The expression of VEGF, integrin β3 protein and mRNA in endometrium were detected by histochemical staining and real-time PCR. The rest of the mice were killed by pd8 to calculate implantation rate. Results: Compared with the control group, the implantation rate and the expression of VEGF and integrin β3 protein and mRNA were significantly lower in the BID group in the implantation window period(P<0.01);the pregnancy rate and the expression of VEGF and integrin β3 protein and mRNA in PBMCs treatment group were significantly higher than these in the BID group(P<0.01),and were close to the level of the control group(P>0.05).Conclusion:PBMCs intrauterine injection can improve the pregnancy rate of BID mice,and its mechanism may be related to the increase of VEGF and integrinβ3 expression in the endometrium of implantation window period,so as to improve the endometrial receptivity of mice.

关 键 词：外周血单个核细胞 子宫内膜容受性 血管内皮生长因子 整合素Β3 小鼠 

分 类 号：R711.6[医药卫生—妇产科学]