催产素诱导大鼠骨髓间充质干细胞分化为心肌样细胞的研究

基础医学与临床 ›› 2011, Vol. 31 ›› Issue (11): 1273-1277.

催产素诱导大鼠骨髓间充质干细胞分化为心肌样细胞的研究

王海萍¹,王浩宇¹,吕洋²,张雷³,陈炜⁴,邵素霞⁵

1. 河北北方学院
2. 河北北方学院组胚教研室
3. 河北医科大学
4. 河北医科大学组织学与胚胎学教研室
5. 河北医科大学组织学胚胎学教研室

收稿日期:2011-05-06 修回日期:2011-07-03 出版日期:2011-11-05 发布日期:2011-11-02
通讯作者: 张雷 E-mail:zhanglei@hebeimu.edu.cn
基金资助:
骨髓间充质干细胞体外诱导向心肌细胞分化及细胞移植治疗心肌梗死的调控机制

Oxytocin induced rat bone marrow mesenchymal stem cells differentiate into cardiomyocyte-like cells in vitro

Received:2011-05-06 Revised:2011-07-03 Online:2011-11-05 Published:2011-11-02

摘要/Abstract

摘要： [摘要] 目的探索催产素（Oxytocin，OT）体外诱导骨髓间充质干细胞（marrow mesenchymal stem cells， MSCs）向心肌细胞分化的可行性。方法取SD大鼠四肢骨骨髓，分离培养MSCs，应用OT定向诱导，相差显微镜观察细胞形态学变化，应用免疫细胞化学、激光扫描共聚焦显微镜技术检测结蛋白（desmin），α-横纹肌肌动蛋白（α-sarcomeric actin）、心肌特异性肌钙蛋白（C-TnT）的表达；在诱导后7d、21d和28d，3个时间点以半定量RT-PCR方法检测细胞心肌早期转录因子GATA-4和心肌特异性肌凝蛋白重链α-MHC的表达情况。结果体外分离培养的细胞生长稳定，增殖较快，主要为纺锤形和成纤维细胞状。:诱导后1周，细胞以梭形、柱状为主，部分细胞有分支, 核卵圆形、居中, 类似心肌细胞形态，排列方向渐趋一致。诱导后4周，细胞形态变长、增大，多呈杆状，紧密平行排列生长。分化后细胞desmin、α-sarcomeric actin 和C-TnT均呈现阳性表达。激光共聚焦扫描显微镜观察：可见细胞质内desmin的表达呈红色，cTnT的表达呈绿色，当两者同时被观察时可见其共表达的部位变成黄色。RT-PCR结果显示，GATA-4于诱导后7d弱表达，21d表达增强，28d表达减弱。α-MHC在诱导后7d不表达，21d弱表达，28d表达明显。结论骨髓间充质干细胞在OT诱导下可定向分化为心肌样细胞。

关键词: 骨髓间充质干细胞, 催产素, 心肌细胞, 细胞分化, 细胞培养, 大鼠

Abstract: [Abstract] Objective To investigate the feasibility of inducing rat bone marrow mesenchymal stem cells(MSCs) differentiating into cardiomyocytes by Oxytocin(OT). Methods SD rat MSCs were isolated and cultured from rat bone marrow, then induced by OT for 72 h. The cultured cells were observed by phase-contrast microscope. The laser scanning confocal microscope (LSCM) was used for the expression of desmin, α-sarcomeric actin and C-TnT. GATA-4 and α-MHC expression was detected by relative quantitative RT-PCR after 7, 21, 28 days of induction respectively. Results The cultured cells grow stablely and proliferate faster, mainly appeared as fusiform and fibroblastic shape. After induced by OT for one week, most cells became spindle-like and pillar-form with oval nuclei lying in the center of cells and similar to cardiomyocytes. The direction of the cell arraying was similar gradually. After four weeks, a majority of cells enlarged,lengthened，showed stick-like shape and paralleled closely.The direction of the cell arraying was similar gradually. MSCs induced by OT could be identified by the positive staining for desmin, α-sarcomeric actin and C-TnT. laser scanning confocal microscopy indicated that desmin(α-sarcomeric actin ) was signed red, cTnT was green.When they were both been showed,it changed to yellow.RT-PCR assessment showed that the differentiated cells began to express GATA-4 from day 7 to day 28 of differentiation and began to express α-MHC from day 21 to day 28 of differentiation. Conclusion　MSCs can differentiate into cardiomyocytes in vitro after induced by OT.

Key words: marrow mesenchymal stem cells, Oxytocin, cardiomyocytes, differentiation, cell culture, rat

王海萍王浩宇吕洋张雷陈炜邵素霞. 催产素诱导大鼠骨髓间充质干细胞分化为心肌样细胞的研究[J]. 基础医学与临床, 2011, 31(11): 1273-1277.

[1]	李贵明王彪方蒡何旭李燕. 谷氨酰胺上调2型糖尿病大鼠结肠组织GLP-1表达[J]. 基础医学与临床, 2019, 39(6): 887-889.
[2]	张源洲席雨鑫温馨李鸿珠. 硫化氢对衰老缺血心肌细胞保护作用的研究进展[J]. 基础医学与临床, 2019, 39(2): 252-255.
[3]	赵承承陈建泉吕纯业. 转染miRNAs降低脂多糖诱导的大鼠肠微血管内皮细胞水通道蛋白1表达[J]. 基础医学与临床, 2019, 39(1): 42-46.
[4]	陈晓依王浩宇吕洋王海萍孙微李柔. PFT-α与BMP-2联合诱导大鼠BMMSCs分化为心肌样细胞[J]. 基础医学与临床, 2018, 38(8): 1088-1093.
[5]	任春贞骆亚莉刘永琦李程豪张艳辉王风梅卢志伟李玲. IL-6和TNF-α模拟炎性微环境促进人BMSCs向肿瘤相关成纤维细胞分化[J]. 基础医学与临床, 2018, 38(7): 901-906.
[6]	胡慧吴强田茂波. 转染携带Ifi204基因的慢病毒可上调大鼠主动脉成纤维细胞的p204表达[J]. 基础医学与临床, 2018, 38(2): 158-162.
[7]	吴庆程阔菊姚浩旗黄河杨天德. 芒果苷对原代大鼠心肌细胞缺血缺氧凋亡的影响[J]. 基础医学与临床, 2018, 38(2): 169-173.
[8]	角建林唐薇郑红赵宏斌. 彝药恒古骨伤愈合剂缓解去势树鼩骨质疏松症[J]. 基础医学与临床, 2018, 38(12): 1686-1690.
[9]	周芳亮何迎春闫庆梓张凯强廖端芳. 大鼠骨髓间充质干细胞向血管平滑肌细胞分化过程中小窝蛋白-1表达增强[J]. 基础医学与临床, 2018, 38(11): 1517-1521.
[10]	赖星宇朱静田杰易勤陈雪妮. Akt在Islet-1促C3H10T1/2向心肌细胞特异分化过程中的作用[J]. 基础医学与临床, 2018, 38(1): 13-19.
[11]	崔桂丽王瑞玲马琴闫佳敏王亚静. 自噬在高糖高脂引起H9C2心肌细胞系凋亡中的影响[J]. 基础医学与临床, 2017, 37(12): 1699-1705.
[12]	王记陆玉琴徐鑫纪召娟. 抑瘤素M促进小鼠诱导多能干细胞向心肌分化[J]. 基础医学与临床, 2017, 37(10): 1401-1406.
[13]	刘振李硕刘玲玲郭志坤王鹏飞. c-kit与microRNA-21在心衰大鼠心肌中表达下降[J]. 基础医学与临床, 2017, 37(1): 76-79.
[14]	王越朱静田杰许皓易勤赖星宇. Islet-1诱导成纤维细胞C3H10T1/2向心肌细胞分化过程中组蛋白乙酰化与DNA甲基化在GATA4启动子区的关系[J]. 基础医学与临床, 2016, 36(9): 1179-1186.
[15]	李丽娜孙伟铭高君王跃虹魏璨徐长庆李鸿珠. 外源性H2S恢复老龄大鼠心肌缺血后适应对I/R损伤的缓解作用[J]. 基础医学与临床, 2016, 36(9): 1199-1205.