摘要
Objective:To investigate the effect of stromal interaction molecule 1（STIM1） knockdown on the proliferation and migration of endothelial progenitor cells（EPCs） after vascular injury and its mechanism.Methods:The rat bone marrow derived EPCs were divided into three groups:adenovirus negative control（group NSC）,rat STIM1 adenovirus vector transfection group（group si/rSTIM1） and rat &human recombinant STIM1 adenovirus transfection group（group si/rSTlM1+hSTIM1）.The STIM1 expressions in each group were detected by reverse transcription PCR after transfection;the cell proliferation was tested by[<sup>3</sup>H]thymidine incorporation assay（<sup>3</sup>HTdR）;Cell cycle was analyzed by flow cytometry;the cells’ migration activity was detected by Boyden assay;Calcium ion concentration was detected by using laser confocal method.Results:48 h later after transfection,the expression level of STIM1 in si/rSTIM1 cells was significantly lower than that in NSC group（0.21±0.12 vs 1.01±0.01,P【0.05）;EPCs that stayed in G<sub>1</sub> phase in si/rSTIM1 group[（93.3l±0.24）%]were significantly more than that in NSC group[（78.03±0.34）%,P【0.05];EPCs’ migration activity in si/rSTIM1 group（10.03±0.33） was significantly lower than that in NSC group:（32.11±0.54,P【0.05）;EPCs calcium ion concentration changes in EPCs in si/rSTIM1group（38.03±0.13） was significantly lower than that in NSC group（98.11±0.34,P【0.05）.While there was no significant difference between si/rSTIM1+hSTIM1 group and NSC group on the four indexes above.Conclusions:Silence of STIM1 attenuates EPCs proliferation and migration after vascular injury,by mediating the calcium ion concentration in EPCs.
Objective:To investigate the effect of stromal interaction molecule 1（STIM1） knockdown on the proliferation and migration of endothelial progenitor cells（EPCs） after vascular injury and its mechanism.Methods:The rat bone marrow derived EPCs were divided into three groups:adenovirus negative control（group NSC）,rat STIM1 adenovirus vector transfection group（group si/rSTIM1） and rat &human recombinant STIM1 adenovirus transfection group（group si/rSTlM1+hSTIM1）.The STIM1 expressions in each group were detected by reverse transcription PCR after transfection;the cell proliferation was tested by[3H]thymidine incorporation assay（3HTdR）;Cell cycle was analyzed by flow cytometry;the cells’ migration activity was detected by Boyden assay;Calcium ion concentration was detected by using laser confocal method.Results:48 h later after transfection,the expression level of STIM1 in si/rSTIM1 cells was significantly lower than that in NSC group（0.21±0.12 vs 1.01±0.01,P<0.05）;EPCs that stayed in G1 phase in si/rSTIM1 group[（93.3l±0.24）%]were significantly more than that in NSC group[（78.03±0.34）%,P<0.05];EPCs’ migration activity in si/rSTIM1 group（10.03±0.33） was significantly lower than that in NSC group:（32.11±0.54,P<0.05）;EPCs calcium ion concentration changes in EPCs in si/rSTIM1group（38.03±0.13） was significantly lower than that in NSC group（98.11±0.34,P<0.05）.While there was no significant difference between si/rSTIM1+hSTIM1 group and NSC group on the four indexes above.Conclusions:Silence of STIM1 attenuates EPCs proliferation and migration after vascular injury,by mediating the calcium ion concentration in EPCs.
基金
supported by Project of National Health and Family Planning Commission in Pudong New Area of Shanghai(PW2013A-35)
Special Industrial Project of National Health and Family Planning Commission in Pudong New Area of Shanghai(PW2013E-2)
Key Project of Health Bureau of Shanghai(20124022)
