中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
8期
1687-1689
,共3页
孙永丰%冯剑锷%史嘉玮%董念国
孫永豐%馮劍鍔%史嘉瑋%董唸國
손영봉%풍검악%사가위%동념국
S24F基因%调节活化正常T细胞表达和分泌因子%腺病毒载体%基因转染
S24F基因%調節活化正常T細胞錶達和分泌因子%腺病毒載體%基因轉染
S24F기인%조절활화정상T세포표체화분비인자%선병독재체%기인전염
S24F gene%Regulated upon activation normal T cell expressed and secreted%Adenovirus vector%Gene transfer
目的 观察腺病毒介导S24F基因转染人脐静脉内皮细胞(HUVECs)对调节活化正常T细胞表达和分泌因子(RANTES)趋化作用的影响.方法 构建pAV-MCMV/S24F-GFP-3 FLAG(Ad-S24F)腺病毒载体,将不含目的基因的病毒载体pAV-MCMV-GFP (Ad-Null)为阴性对照组.分离培养HUVECs后分别转染Ad-S24F和Ad-Null,另设不含腺病毒培养基为空白对照.转染后采用荧光显微镜及Western blot检测重组蛋白表达.采用Transwell小室法分析S24F对RANTES趋化作用的影响.结果 Ad-S24F及Ad-Null构建成功,转染HUVECs后荧光显微镜及Western blot能检测到重组蛋白表达,Ad-S24F转染后能够抑制RANTES诱导的外周血单个核细胞(PBMCs)穿透内皮[Ad-S24F:(9.20 ±0.02)%;Ad-Null:(17.70±0.02)%;空白对照组(15.10±0.01)%]的趋化作用.结论 腺病毒介导S24F基因转染HUVECs能够抑制RANTES的趋化作用.
目的 觀察腺病毒介導S24F基因轉染人臍靜脈內皮細胞(HUVECs)對調節活化正常T細胞錶達和分泌因子(RANTES)趨化作用的影響.方法 構建pAV-MCMV/S24F-GFP-3 FLAG(Ad-S24F)腺病毒載體,將不含目的基因的病毒載體pAV-MCMV-GFP (Ad-Null)為陰性對照組.分離培養HUVECs後分彆轉染Ad-S24F和Ad-Null,另設不含腺病毒培養基為空白對照.轉染後採用熒光顯微鏡及Western blot檢測重組蛋白錶達.採用Transwell小室法分析S24F對RANTES趨化作用的影響.結果 Ad-S24F及Ad-Null構建成功,轉染HUVECs後熒光顯微鏡及Western blot能檢測到重組蛋白錶達,Ad-S24F轉染後能夠抑製RANTES誘導的外週血單箇覈細胞(PBMCs)穿透內皮[Ad-S24F:(9.20 ±0.02)%;Ad-Null:(17.70±0.02)%;空白對照組(15.10±0.01)%]的趨化作用.結論 腺病毒介導S24F基因轉染HUVECs能夠抑製RANTES的趨化作用.
목적 관찰선병독개도S24F기인전염인제정맥내피세포(HUVECs)대조절활화정상T세포표체화분비인자(RANTES)추화작용적영향.방법 구건pAV-MCMV/S24F-GFP-3 FLAG(Ad-S24F)선병독재체,장불함목적기인적병독재체pAV-MCMV-GFP (Ad-Null)위음성대조조.분리배양HUVECs후분별전염Ad-S24F화Ad-Null,령설불함선병독배양기위공백대조.전염후채용형광현미경급Western blot검측중조단백표체.채용Transwell소실법분석S24F대RANTES추화작용적영향.결과 Ad-S24F급Ad-Null구건성공,전염HUVECs후형광현미경급Western blot능검측도중조단백표체,Ad-S24F전염후능구억제RANTES유도적외주혈단개핵세포(PBMCs)천투내피[Ad-S24F:(9.20 ±0.02)%;Ad-Null:(17.70±0.02)%;공백대조조(15.10±0.01)%]적추화작용.결론 선병독개도S24F기인전염HUVECs능구억제RANTES적추화작용.
Objective To investigate the effects of chemoattractant function of regulated upon activation normal T cell expressed and secreted (RANTES) with adenovirus-mediated gene transfer of S24F of the human umbilical vein endothelial cells (HUVECs).Methods Construction of the recombinant Adenoviruses Vector pAV-MCMV/S24F-GFP-3FLAG (Ad-S24F),an adenoviral vector containing no transgene pAV-MCMV-GFP (Ad-Null) was used as a control.HUVECs were isolated and cultured in vitro,and were transfected with Ad-S24F (Ad-S24F group),Ad-Null (Ad-Null group),or transfected with no virus (control group).The expression of reconstructive protein after transfection was detected by using Western blotting and fluorescence microscopy in each group.The in vitro transendothelialchemotaxis assay was used to compare RANTES-induced transmigration of peripheral blood mononuclear cells (PBMCs) across HUVECs cultured on the upper Transwell chamber.Results The successful construction of recombinant adenoviral vector carrying S24F gene.RANTES-induced PBMC transendothelialchemotaxis is inhibited by S24F [Ad-S24F:(9.20 ± 0.02) % ; Ad-Null:(17.70 ± 0.02) % ; control:(15.10 ± 0.01) %].Conclusion Adenovirus-mediated gene transfer of S24F of the human umbilical vein endothelial cells may inhibit RANTES-induced PBMC transendothelialchemotaxis.