人胎盘来源间充质干细胞对妊娠期高血压病细胞滋养细胞凋亡的影响：随机对照实验方案

摘要

背景：细胞滋养细胞凋亡增加、浸润能力下降是妊娠期高血压病发病的重要环节之一。人胎盘来源间充质干细胞已经在许多研究中显示出了对损伤的修复作用，而且胎盘间充质干细胞还具有一定的内分泌功能，可通过自分泌或旁分泌细胞因子的方式作用于近、远处的组织。因此，尝试移植人胎盘来源间充质干细胞修复损伤的滋养细胞功能，进而治疗妊娠期高血压病，有一定可行性。　　方法/设计：随机对照细胞学实验。收集人胎盘来源间充质干细胞培养液，过滤后即为人胎盘来源间充质干细胞条件培养基。培养人绒毛膜滋养层细胞系JEG-3细胞，随机分为3组，对照组在培养液中加入体积分数15%正常孕妇血清；妊娠期高血压病模型组在培养液中加入体积分数15%子痫前期重度患者的血清；人胎盘来源间充质干细胞条件培养组在培养液中加入体积分数15%子痫前期重度患者血清培养24 h后，更换为体积分数15%子痫前期重度患者血清+人胎盘来源间充质干细胞条件培养基。采用流式细胞仪检测各组细胞的凋亡率。讨论：实验从人胎盘来源间充质干细胞对妊娠期高血压病细胞滋养细胞功能的影响入手，可能为妊娠期高血压疾病的治疗找到一条细胞移植之路。　　伦理批准：研究方案取得沈阳医学院附属中心医院临床研究伦理委员会的书面批准。正常及子痫前期孕妇签署知情同意书。%BACKGROUND:Excessive apoptosis and decreased infiltration of cytotrophoblasts are essential causes for hypertension in pregnancy. Human placenta-derived mesenchymal stem cels contribute to damage repair, which has been shown in many studies, and moreover, human placenta-derived mesenchymal stem cels have a certain endocrine function and can act on the other tissues in an autocrine or paracrine manner. Therefore, we attempt to explore whether the human placenta-derived mesenchymal stem cel can repair damaged cytotrophoblasts, and then to gestational hypertension. METHODS/DESIGN:This is a randomized controled cytological experiment. Human placenta-derived mesenchymal stem cel culture medium is colected and filtrated as human placenta-derived mesenchymal stem cel conditioned medium. Human cytotrophoblasts, JEG-3 cels, are cultured and randomized into three groups:15% normal pregnant serum is added in control group; 15% serum from severe pre-eclampsia patients is added in gestational hypertension model group; and in condition medium group, 15% serum from severe pre-eclampsia patients is added for 24 hours of culture, and then human placenta-derived mesenchymal stem cel conditioned medium containing 15% serum from severe pre-eclampsia patients is used instead. Flow cytometry is used to detect cel apoptosis rate. DISCUSSION: This study wil help to find the feasibility of cel transplantation for gestational hypertension by exploring the effect of human placenta-derived mesenchymal stem cels on cytotrophoblasts function in gestational hypertension. ETHICAL APPROVAL: The protocol is approved by the Ethics Committee of Shenyang Central Hospital of Shenyang Medical University. Informed consent is obtained from normal and pre-eclampsia women in pregnancy.