S1P通过Cx43对成肌细胞移植治疗心肌梗死后心律失常的作用和机制研究

项目名称： S1P通过Cx43对成肌细胞移植治疗心肌梗死后心律失常的作用和机制研究

项目编号： No.81460048

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 于欢

作者单位： 九江学院

项目金额： 49万元

中文摘要： 结合大量文献和坚实的前期基础，延续前一个国家基金课题，以心肌电均一性为视角，沿着1-磷酸鞘氨醇(S1P)借助Ca2+通过NO、miR-1借导的通路调控缝隙连接蛋白Cx43(Cx43)作用于移植区细胞，减少成肌细胞移植治疗心肌梗死后心律失常发生的思路，朝着Cx43调节心肌电均一性减少心律失常的目标开展工作。以在体心肌梗死动物模型和离体成肌细胞和心肌细胞共培养模型为研究对象，采用心电图、射频记录、透射电镜、钙离子荧光探针负载、膜片钳技术（原位、全细胞检测）等针对性手段，并结合RT-PCR、Western blot等常规方法检测心律失常发生率、移植细胞与宿主细胞之间的功能耦合、细胞电生理特性、Ca2+浓度、SERCA2a表达、NO含量、miR-1表达、Cx43 mRNA和蛋白表达及磷酸化水平，探讨S1P减少移植后心律失常的作用及分子机制，为减少自体成肌细胞移植后心律失常发生提供新思路和新方法。

中文关键词： 1-磷酸鞘氨醇；成肌细胞；心律失常；钙离子；缝隙连接蛋白Cx43

英文摘要： This research is the continuity of previous project funded by National Natural Science Foundation, based on previous studies and our research, connexin 43 (Cx43) regulates myocardial electrical homogeneity to reduce cardiac arrhythmia. Our hypothesis is that sphingosine 1-phosphate (S1P) regulates Cx43 through Ca2+，SERCA2a and microRNA-1 (miR-1), decreased cardiac arrhythmia after myoblast transplantation on myocardial infarction (MI). For the prevention and control of the arrhythmia after myoblasts transplantation, using animal model of myocardial infarction in vivo, we transfer S1P into the myoblasts. Then cells were transplanted to animal model of MI. In vivo, using ECG, radio frequency, transmission electron microscopy, the patch clamp technique，calcium fluorescent probe, and nitrate reductase immunohistochemistry, RT-PCR, Western-blotting, immunoperoxidase staining, observe the incidence of arrhythmia, measure the function of the coupling between the transplanted cells and host cells, detect electrophysiological characteristics of cells, check space structure and phosphorylation level of Cx43, measure expression of miR-1 and NO of myocardial tissue. In vitro, we use coculture model of myoblasts and cardiomyocytes, by transmission electron microscopy, calcium fluorescent probe, and nitrate reductase immunohistochemistry, RT-PCR, Western-blot and the patch clamp technique detect the connections of two cells, check electrophysiological characteristics of cells,, measure expression of miR-1. Try to find the role and molecular mechanism of S1P on arrhythmia after myoblast transplantation treatment of MI. In order to provide new ideas and new methods of decreasing arrhythmia after myoblasts autotransplantation. It will provide a theoretical basis for myoblasts autotransplantation on treatment of MI.

英文关键词： S1P;myoblast;arrhythmia;Ca2+;Cx43