分化抑制因子1对创伤组织新生血管化的促进作用观察  

Studies on inhibitor of differentiation 1 in promoting neovascularization post trauma

作　　者：孙荣距[1] 张建波[1] 果应菲[1] 秦宇红[1] 党伟[1] 张宪[1] 袁晓玲[1] 赵晓东[1] 

机构地区：[1]解放军总医院第一附属医院急救部,北京100048

出　　处：《解放军医学杂志》2010年第3期285-288,共4页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金青年基金(30700869)

摘　　要：目的探讨分化抑制因子1(Id1)对创伤组织新生血管化的促进作用及其机制。方法经基因鉴定的SD大鼠16只,随机分为非创伤组(即正常对照组,n=4)和创伤组(包括Id1基因敲除组、Id1诱导表达组和Id1正常表达组,n=4)。采用MTT法测定Id1对血管内皮细胞Eahy926增殖的影响,实时定量RT-PCR、蛋白免疫印迹技术检测创伤1、24、48、72h后创伤组织中新生血管化标志物血管内皮细胞生长因子(VEGF)、CD105、内皮素-1(ET-1)等的mRNA和蛋白表达变化,并用报告基因技术检测Id1对VEGF、CD105、ET-1基因表达的调控作用。结果血管内皮细胞模型实验显示转染Id1siRNA后,血管内皮细胞Eahy926增殖明显受到抑制,以72h时最显著(P<0.01);而转染pcDNAId1后,细胞增殖于48h时即增强(P<0.05),以72h时最显著(P<0.01)。创伤组Id1基因敲除大鼠VEGF、CD105的mRNA及蛋白表达均受到抑制(P<0.05),且伤后各时间点表达水平无明显变化。Id1诱导表达组24h后大鼠Id1、VEGF、CD105表达均明显增强(P<0.05),于72h达高峰(P<0.01),而ET-1mRNA水平在伤后24h升高,48h达高峰(P<0.05),72h下降。报告基因检测结果显示,Id1敲除后VEGF、CD105荧光素酶活性明显受到抑制(P<0.05),而Id1基因诱导表达后VEGF、CD105活性明显增强(P<0.01),但Id1对ET-1的表达无明显影响。结论Id1能有效促进血管内皮细胞Eahy926增殖,调节血管内皮细胞、创伤组织中VEGF、CD105以及ET-1的表达,在组织细胞新生血管化中起重要作用。Objective To explore the effect and mechanism of inhibitor of differentiation 1 （Id1） on the promoting of neovascularization post trauma. Methods Sixteen SD rats were randomly divided into non-trauma group （NT group, n=4） and trauma group including Id1 knocking-out group, Id1 over-expressing group and Id1 normal group （n=4）. The influence of Id1 on the Eahy926 cell proliferation was detected by MTT. Targeted marker proteins including Id1, vascular endothelial growth factor （VEGF）, endoglin（CD105）, and ET-1 were detected at 1, 24, 48h and 72h post trauma by real-time quantitative RT-PCR and Western blotting. The effect of Id1 on VEGF, CD105 and ET-1 was detected by luciferase report gene assay. Results The experiment of vascular endothelial cells model showed that when transfected with Id1siRNA, the cells proliferation of Eahy926 were significantly inhibited, especially at 72h post trauma （P0.01）, while promoted at 48h （P0.05） and 72h （P0.01）when pcDNAId1 vectors was transfected. The VEGF and CD105 expression of rats in Id1 knocking-out group was inhibited in mRNA and protein levels （P0.05）. No changes were found at different time points post trauma. The expression of Id1, VEGF and CD105 all increased at 24h post trauma （P0.05）and reached the peak level at 72h （P0.01） when Id1 was over expressed. While the ET-1 increased at 24h. reached the peak level at 48h （P0.05）and decreased at 72h. Luciferase report assay showed that the luciferase activity was inhibited remarkably after the Id1was knocked out （P0.05）, while increased when Id1 was over expressed （P0.01）. Id1 showed no effect on ET-1 expression. Conclusions Id1 could efficiently promote Eahy926 cell proliferation and the expression of VEGF and CD105 post trauma. It is possibly that Id1 plays an important role on the neovascularization and trauma reparation by regulating the expression of VEGF and CD105.

关 键 词：基因 Id1 创伤和损伤 新生血管化 病理性 

分 类 号：R641.023[医药卫生—外科学]