摘要： 目的 评估丹参酮ⅡA对人胎盘间充质干细胞（hPDMSCs）向心肌细胞分化及预分化细胞归巢能力的影响，并探讨其相关机制。方法 应用组织块法分离培养hPDMSCs。取第3~6代对数生长期的hPDMSCs，分别用丹参酮ⅡA或丹参酮ⅡA+β-catenin激动剂WAY-262611处理，同时设置不加药的对照组，20 d终止实验。用MTS法评估细胞增殖情况，划痕实验分析预分化细胞向损伤区域迁移情况，Western blotting检测心肌特异性蛋白（GATA-4、ANP、α-SCA、cTnI）、Wnt/β-catenin通路相关蛋白（APC、Gsk-3β、β-catenin）以及与归巢相关的趋化因子（CXCR-4、CCR-2）的表达情况。结果 应用组织块法成功分离培养出hPDMSCs；与对照组相比，经丹参酮ⅡA干预的hPDMSCs增殖速率明显减慢，心肌特异性蛋白GATA-4、ANP、α-SCA、cTnI的表达明显增加（P<0.05），向损伤区迁移的能力明显增强，且在这个过程中，APC、Gsk-3β、CXCR-4、CCR-2的表达明显增多（P<0.05），β-catenin的表达明显减少（P<0.05）。但当丹参酮ⅡA与β-catenin激动剂WAY-262611联合干预hPDMSCs后，丹参酮ⅡA抑制细胞增殖、促进心肌特异性蛋白GATA-4、ANP、α-SCA、cTnI的表达、提高细胞向损伤区迁移的能力明显降低，且CXCR-4、CCR-2的表达也明显减少（P<0.05）。结论 丹参酮ⅡA能够通过调控Wnt/β-catenin通路诱导hPDMSCs向心肌细胞分化、提高预分化细胞向损伤区的归巢能力。

丹参酮ⅡA诱导人胎盘间充质干细胞向心肌细胞分化、提高预分化细胞归巢能力的研究

樊晨星¹, 王秀艳¹, 赵千², 赖红³, 李昆¹

1. 锦州医科大学附属第一医院检验科, 辽宁 锦州 121001;
2. 锦州医科大学附属第三医院检验科, 辽宁 锦州 121001;
3. 中国医科大学基础医学院解剖学教研室, 沈阳 110122

收稿日期:2019-05-21出版日期:2020-04-30发布日期:2020-04-18
通讯作者:李昆E-mail:lklszlym@163.com
作者简介:樊晨星(1995-),女,硕士研究生.
基金资助:国家自然科学基金（81303255）；辽宁省自然科学基金（2019-MS-142）


关键词: 丹参酮ⅡA, 人胎盘间充质干细胞, 心肌分化, 细胞归巢
Abstract: Objective To evaluate the effect of tanshinone ⅡA (Tan ⅡA) in the differentiation of human placenta-derived mesenchymal stem cells (hPDMSCs) into cardiomyocytes,the homing of pre-differentiated cells,and to investigate the related underlying mechanisms. Methods The hPDMSCs were isolated using an explant culture method. The third to the sixth generation hPDMSCs were treated with Tan ⅡA or Tan ⅡA combined with the β-catenin agonist WAY-262611,and a control group was established. The MTS method was used to measure cell proliferation,and a scratch test was used to analyze the migration of pre-differentiated cells to the injured area. Western blotting was used to detect the expression of myocardial-specific proteins (GATA-4,ANP,α-SCA,and cTnI),Wnt/β-catenin pathway-related proteins (APC,Gsk-3β,and β-catenin),and homing-associated chemokines (CXCR-4 and CCR-2). Results Using the explant culture method,hPDMSCs were successfully isolated. Compared with the control group,the proliferation of cells treated with TanⅡA decreased significantly;the expression of GATA-4,ANP,α-SCA,and cTnI were significantly increased (P<0.05),and the ability of pre-differentiated cells to migrate to the injured area were significantly enhanced. In this process,the expression of APC,Gsk-3β,CXCR-4,and CCR-2 were significantly increased (P<0.05),and the expression of β-catenin was significantly decreased (P<0.05). However,co-treatment with the β-catenin agonist WAY-262611 reverted the aforementioned actions of Tan ⅡA. Conclusion Tan ⅡA can induce differentiation of hPDMSCs into cardiomyocytes and enhance the homing of pre-differentiated cells to injured areas by regulating the Wnt/β-catenin pathway.
Key words: tanshinone ⅡA, human placental mesenchymal stem cells, myocardial differentiation, cell homing
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