中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2015年
2期
157-161
,共5页
王温慧%李新明%柳亮%侯佳音%朱茜%丛欣鹏
王溫慧%李新明%柳亮%侯佳音%硃茜%叢訢鵬
왕온혜%리신명%류량%후가음%주천%총흔붕
内皮,血管%干细胞%细胞增殖%细胞运动%沉默基质交感分子1
內皮,血管%榦細胞%細胞增殖%細胞運動%沉默基質交感分子1
내피,혈관%간세포%세포증식%세포운동%침묵기질교감분자1
Endothelium,vascular%Stem cells%Cell proliferation%Cell movement%Stromal interaction molecule 1
目的 探讨沉默基质交感分子1(STIM1)对鼠内皮祖细胞(EPC)增值及迁移的影响,并探讨其机制.方法 取鼠骨髓源的EPC,分为3组,即腺病毒阴性对照(NSC组)、大鼠STIM1腺病毒载体转染组(Ad-si/rSTIM1组)和大鼠及人重组STIM1腺病毒共转染组(Ad-si/rSTIM1+hSTIM1组).转染后,采用逆转录聚合酶链反应检测STIM1的表达水平,[3H]胸腺嘧啶核苷掺入法(3H-TdR)检测细胞的增殖情况,流式细胞仪分析细胞周期分布,Boyden小室检测细胞迁移数,激光共聚焦法检测钙离子浓度变化.结果 细胞转染后48 h,Ad-si/rSTIM1组EPC内STIM1 mRNA的表达水平低于NSC组(0.21 ±0.12比1.01±0.01,P＜0.05),Ad-si/rSTIM1组EPC的3H-TdR掺入量低于NSC组(P=0.001),Ad-si/rSTIM1+ hSTIM1组EPC的3 H-TdR掺入量与NSC组比较差异无统计学意义.Ad-si/rSTIM1组EPC周期停止于G1期的细胞数比率为(93.31±0.24)％高于NSC组的(78.03±0.34)％(P＜0.05).Ad-si/rSTIM1组EPC迁移数为(10.03±0.33)个低于NSC组的(32.11 ±0.54)个(P＜0.05).Ad-si/rSTIM1组EPC内钙离子浓度变化为(38.03 ±0.13)a.u.小于NSC组的(98.11 ±0.34)a.u.(P＜0.05).而Ad-si/rSTIM1+ hSTIM1组EPC内STIM1的mRNA表达水平、细胞迁移活动、钙离子浓度变化及停留在G1期的细胞数与NSC组比较差异均无统计学意义.结论 沉默STIM1可抑制血管损伤后EPC的增殖和迁移,STIM1对EPC的调节作用主要通过对细胞内外钙离子水平的调控实现.
目的 探討沉默基質交感分子1(STIM1)對鼠內皮祖細胞(EPC)增值及遷移的影響,併探討其機製.方法 取鼠骨髓源的EPC,分為3組,即腺病毒陰性對照(NSC組)、大鼠STIM1腺病毒載體轉染組(Ad-si/rSTIM1組)和大鼠及人重組STIM1腺病毒共轉染組(Ad-si/rSTIM1+hSTIM1組).轉染後,採用逆轉錄聚閤酶鏈反應檢測STIM1的錶達水平,[3H]胸腺嘧啶覈苷摻入法(3H-TdR)檢測細胞的增殖情況,流式細胞儀分析細胞週期分佈,Boyden小室檢測細胞遷移數,激光共聚焦法檢測鈣離子濃度變化.結果 細胞轉染後48 h,Ad-si/rSTIM1組EPC內STIM1 mRNA的錶達水平低于NSC組(0.21 ±0.12比1.01±0.01,P＜0.05),Ad-si/rSTIM1組EPC的3H-TdR摻入量低于NSC組(P=0.001),Ad-si/rSTIM1+ hSTIM1組EPC的3 H-TdR摻入量與NSC組比較差異無統計學意義.Ad-si/rSTIM1組EPC週期停止于G1期的細胞數比率為(93.31±0.24)％高于NSC組的(78.03±0.34)％(P＜0.05).Ad-si/rSTIM1組EPC遷移數為(10.03±0.33)箇低于NSC組的(32.11 ±0.54)箇(P＜0.05).Ad-si/rSTIM1組EPC內鈣離子濃度變化為(38.03 ±0.13)a.u.小于NSC組的(98.11 ±0.34)a.u.(P＜0.05).而Ad-si/rSTIM1+ hSTIM1組EPC內STIM1的mRNA錶達水平、細胞遷移活動、鈣離子濃度變化及停留在G1期的細胞數與NSC組比較差異均無統計學意義.結論 沉默STIM1可抑製血管損傷後EPC的增殖和遷移,STIM1對EPC的調節作用主要通過對細胞內外鈣離子水平的調控實現.
목적 탐토침묵기질교감분자1(STIM1)대서내피조세포(EPC)증치급천이적영향,병탐토기궤제.방법 취서골수원적EPC,분위3조,즉선병독음성대조(NSC조)、대서STIM1선병독재체전염조(Ad-si/rSTIM1조)화대서급인중조STIM1선병독공전염조(Ad-si/rSTIM1+hSTIM1조).전염후,채용역전록취합매련반응검측STIM1적표체수평,[3H]흉선밀정핵감참입법(3H-TdR)검측세포적증식정황,류식세포의분석세포주기분포,Boyden소실검측세포천이수,격광공취초법검측개리자농도변화.결과 세포전염후48 h,Ad-si/rSTIM1조EPC내STIM1 mRNA적표체수평저우NSC조(0.21 ±0.12비1.01±0.01,P＜0.05),Ad-si/rSTIM1조EPC적3H-TdR참입량저우NSC조(P=0.001),Ad-si/rSTIM1+ hSTIM1조EPC적3 H-TdR참입량여NSC조비교차이무통계학의의.Ad-si/rSTIM1조EPC주기정지우G1기적세포수비솔위(93.31±0.24)％고우NSC조적(78.03±0.34)％(P＜0.05).Ad-si/rSTIM1조EPC천이수위(10.03±0.33)개저우NSC조적(32.11 ±0.54)개(P＜0.05).Ad-si/rSTIM1조EPC내개리자농도변화위(38.03 ±0.13)a.u.소우NSC조적(98.11 ±0.34)a.u.(P＜0.05).이Ad-si/rSTIM1+ hSTIM1조EPC내STIM1적mRNA표체수평、세포천이활동、개리자농도변화급정류재G1기적세포수여NSC조비교차이균무통계학의의.결론 침묵STIM1가억제혈관손상후EPC적증식화천이,STIM1대EPC적조절작용주요통과대세포내외개리자수평적조공실현.
Objective The purpose of this study is to explore the impact of stromal interaction molecule 1 (STIM1) knockdown on the proliferation and migration capacities of endothelial progenitor cells (EPCs).Methods The rat bone marrow derived EPCs were obtained and divided into three groups:adenovirus negative control (NSC) group,rat STIM1 adenovirus vector transfection (si/rSTIM1) group and rat and human recombinant STIM1 adenovirus transfection (si/rSTIM1 + hSTIM1) group.The STIM1 expressions in each group were detected by reverse transcription PCR after transfection.The cell proliferation was tested by [3H] thymidine incorporation assay (3 H-TdR).Cell cycle was analyzed by flow cytometry.The cells migration activity was detected by Boyden assay.Calcium ion concentration was detected by confocal laser scanning microscopy.Results 48 h after transfection,the expression level of STIM1 in si/rSTIM1 group was significantly lower than that in NSC group (0.21 ±0.12 vs.1.01 ±0.01,P ＜0.05),and number of EPCs at G1 phase in si/rSTIM1 group ((93.31 ± 0.24)％) was significantly higher than that in NSC group ((78.03 ± 0.34)％,P ＜ 0.05),and 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),and EPCs calcium ion concentration in EPCs in si/rSTIM1 group (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 above four indexes.Conclusion Silencing STIM1 could attenuate EPCs proliferation and migration capacities by modulating the calcium ion concentration in EPCs.
