中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2014年
12期
2195-2200
,共6页
刘俊许%蒋丽%魏香香%牛琮%陈思锋%孟丹
劉俊許%蔣麗%魏香香%牛琮%陳思鋒%孟丹
류준허%장려%위향향%우종%진사봉%맹단
转录因子Bach1%人微血管内皮细胞%血管内皮细胞生长因子%血红素氧合酶1%细胞迁移
轉錄因子Bach1%人微血管內皮細胞%血管內皮細胞生長因子%血紅素氧閤酶1%細胞遷移
전록인자Bach1%인미혈관내피세포%혈관내피세포생장인자%혈홍소양합매1%세포천이
Transcription factor Bach1%Human microvascular endothelial cells%Vascular endothelial growth factor%Heme oxygenase-1%Cell migration
目的：研究转录因子Bach1对人微血管内皮细胞功能的影响。方法：利用小干扰RNA（ small inter-fering RNA，siRNA）细胞转染技术下调内皮细胞Bach1表达；用Matrigel管腔形成实验检测内皮细胞体外血管新生的能力；用Transwell小室法检测细胞迁移；用CCK-8法测定细胞增殖；用实时荧光定量PCR、Western blotting和ELISA法检测细胞中血红素氧合酶1（heme oxygenase 1，HO-1）和血管内皮细胞生长因子（vascular endothelial growth factor， VEGF） mRNA和蛋白的表达情况；用转染报告基因的方法检测VEGF基因的转录活性。结果：下调内皮细胞Bach1表达明显促进人微血管内皮细胞迁移和管腔形成能力，对内皮细胞增殖能力无明显影响；抑制Bach1表达促进内皮细胞HO-1 mRNA和蛋白的表达，增加VEGF转录活性及mRNA和蛋白的表达。结论：抑制转录因子Bach1表达可增加内皮细胞HO-1和VEGF的表达，促进人微血管内皮细胞迁移和管腔形成，提示Bach1是负性调控血管新生的因子。
目的：研究轉錄因子Bach1對人微血管內皮細胞功能的影響。方法：利用小榦擾RNA（ small inter-fering RNA，siRNA）細胞轉染技術下調內皮細胞Bach1錶達；用Matrigel管腔形成實驗檢測內皮細胞體外血管新生的能力；用Transwell小室法檢測細胞遷移；用CCK-8法測定細胞增殖；用實時熒光定量PCR、Western blotting和ELISA法檢測細胞中血紅素氧閤酶1（heme oxygenase 1，HO-1）和血管內皮細胞生長因子（vascular endothelial growth factor， VEGF） mRNA和蛋白的錶達情況；用轉染報告基因的方法檢測VEGF基因的轉錄活性。結果：下調內皮細胞Bach1錶達明顯促進人微血管內皮細胞遷移和管腔形成能力，對內皮細胞增殖能力無明顯影響；抑製Bach1錶達促進內皮細胞HO-1 mRNA和蛋白的錶達，增加VEGF轉錄活性及mRNA和蛋白的錶達。結論：抑製轉錄因子Bach1錶達可增加內皮細胞HO-1和VEGF的錶達，促進人微血管內皮細胞遷移和管腔形成，提示Bach1是負性調控血管新生的因子。
목적：연구전록인자Bach1대인미혈관내피세포공능적영향。방법：이용소간우RNA（ small inter-fering RNA，siRNA）세포전염기술하조내피세포Bach1표체；용Matrigel관강형성실험검측내피세포체외혈관신생적능력；용Transwell소실법검측세포천이；용CCK-8법측정세포증식；용실시형광정량PCR、Western blotting화ELISA법검측세포중혈홍소양합매1（heme oxygenase 1，HO-1）화혈관내피세포생장인자（vascular endothelial growth factor， VEGF） mRNA화단백적표체정황；용전염보고기인적방법검측VEGF기인적전록활성。결과：하조내피세포Bach1표체명현촉진인미혈관내피세포천이화관강형성능력，대내피세포증식능력무명현영향；억제Bach1표체촉진내피세포HO-1 mRNA화단백적표체，증가VEGF전록활성급mRNA화단백적표체。결론：억제전록인자Bach1표체가증가내피세포HO-1화VEGF적표체，촉진인미혈관내피세포천이화관강형성，제시Bach1시부성조공혈관신생적인자。
[ ABSTRACT] AIM:To determine the role of transcription factor Bach1 in the functions of human microvascular en-dothelial cells ( HMVECs ) .METHODS: Bach1 siRNA was transfected into HMVECs to knock down the expression of Bach1.In vitro endothelial cell tube formation assay in Matrigel culture was used as a surrogate assay for angiogenic poten-tial.Migration of HMVECs was determined by using Transwell chambers.Cell proliferation was measured by CCK-8 assay. Real-time PCR, Western blotting, and ELISA were employed to determine mRNA expression and protein level.Reporter as-say was performed to determine vascular endothelial growth factor ( VEGF) transcriptional activity.RESULTS:Knockdown of Bach1 expression in HMVECs led to an increase in the tube formation and increased endothelial cell migration ability, whereas it has little effect on cell proliferation.Bach1 silencing increased the mRNA and protein expression of heme oxygen-ase-1 (HO-1), and enhanced VEGF transcriptional activation, and mRNA and protein expression.CONCLUSION:Bach1 silencing increases HO-1 and VEGF expression, thus promoting the cell migration and tube formation of HMVECs, indicating that Bach1 is a repressor for angiogenesis.
