环磷腺苷葡胺对骨髓间充质干细胞向心肌细胞分化的诱导作用及其机制探讨  

Induction of meglumine adenosine cyclophosphate on bone marrow mesenchymal stem cells differentiating into cardiomyocytes and the mechanism

作　　者：李丹[1] 张金国[1] 尉希清[1] 刘岩松[2] 

机构地区：[1]济宁医学院附属医院,山东济宁272029 [2]山东省肿瘤医院

出　　处：《山东医药》2015年第42期1-3,共3页Shandong Medical Journal

基　　金：山东省医药卫生科技发展计划项目(2013WSB34007)

摘　　要：目的探讨环磷腺苷葡胺(MCA)诱导骨髓间充质干细胞(BMSCs)向心肌细胞分化的机制。方法体外培养BMSCs,将第3代BMSCs接种于96孔板中,分为对照组、MCA组、MCA+LY294002组。MCA组加入1×10-3mmol/L的MCA,MCA+LY294002组加PI3K/AKT信号通路特异性抑制剂LY294002预处理30 min后,再加入1×10-3mmol/L的MCA,两组均诱导3 d后换为普通培养基继续培养。对照组不做任何处理。培养4周后,采用荧光定量PCR法检测心肌特异性基因GATA结合蛋白4(GATA-4)、缝隙连接蛋白43(CX43)mRNA,采用Western blot法检测PI3K/AKT通路相关蛋白PI3K、磷酸化AKT(p-AKT)、磷酸化叉形头转录因子的O亚型(p-Foxo3a)、DOK家族蛋白(DOK5)]的表达。结果 MCA组、MCA+LY294002组GATA-4、CX43 mRNA表达均高于对照组(P均<0.01),MCA+LY294002组GATA-4、CX43 mRNA表达均低于MCA组(P均<0.01)。MCA组PI3K、p-AKT、pFoxo3a、DOK5表达均高于对照组及MCA+LY294002组(P均<0.01)。结论 MCA可诱导BMSCs分化为心肌细胞,其机制可能与激活PI3K/AKT信号通路有关。Objective To investigate the mechanism of meglumine adenosine cyclophosphate（ MCA） inducing the bone marrow mesenchymal stem cells（ BMSCs） differentiating into cardiomyocytes. Methods BMSCs were cultured in vitro,and the third generation of BMSCs were inoculated into 96 hole boards,which were then divided into the control group,MCA group and MCA ＋ LY294002 group. The MCA group was added with 1 × 10- 3mmol / L MCA,the MCA ＋LY294002 group was added with PI3 K / AKT signaling pathway specific inhibitor 30 min pretreated with LY294002,and then was added with 1 × 10- 3mmol / L MCA. After induction for 3 days,the conventional culture medium was used in the two groups to continue the culture. The control group was not treated. After 4 weeks,the mRNA expression of myocardial specific GATA gene binding protein 4（ GATA-4） and gap junction protein 43（ Cx43） was detected by fluorescence quantitative PCR. The expression of PI3 K / AKT pathway related proteins in each group,including phosphatidylinositol 3-kinase（ PI3K）,serine/threonine protein kinase（ Akt）,a forked head transcription factor O subtype（ p-Foxo3a） and the DOK family proteins（ DOK5）,was detected by Western blotting. Results The expression of GATA-4 and CX43 mRNA in the MCA group and MCA ＋ LY294002 group was higher than that of the control group（ all P 0. 01）,the expression of GATA-4 and CX43 mRNA in the MCA ＋ LY294002 group was lower than that in the MCA group（ all P〈 0. 01）. The expression of PI3 K,p-AKT,p-Foxo3 a and DOK5 in the MCA group was higher than that in the control group and MCA ＋LY294002 group（ all P〈 0. 01）. Conclusion MCA can induce BMSCs differentiating into cardiomyocytes,and its mechanism may be related to the activation of PI3 K / AKT signaling pathway.

关 键 词：环磷腺苷葡胺 骨髓间充质干细胞 心肌细胞 细胞分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]