摘要
目的：观察脂质体介导转移的VEGF基因在血管平滑肌细胞（VSMC）中的表达，比较VEG对血管内皮细胞（VEC）及VSMC增殖的影响。方法：将含血管内皮生长因子（VECF）CDNA的真核表达载体pSV121用脂质体介导的基因转移法导入血管平滑肌细胞（VSMC）中，取此VSMC条件培养液，再行血管内皮细胞（VEC）培养，用Northmblot杂交，Westem印迹法及[~3H]胸腺嘧啶核苷掺入法，观察了VEGF在VSMC及其条件培养液中的表达，比较了VEGF对VEC及VSMC增殖的影响。结果：转基因组VSMC中VEGFmRNA呈稳定高表达，转基因组VSMC条件培养液中，VEGF抗原表达显著高于对照组（P均＜0．01），加入此条件培养液的各组VEC的[~3H]胸腺嘧啶核苷掺入量均显著高于对照组（p均＜0．01），而在VSMC组无显著差异。结论：VEGF的转录和表达表明脂质体介导的血管细胞VEGF基因转移是成功的。VEGF具有促VEC增殖作用，但对VSMC无促增殖作用，有利于缺血性疾病及血管再狭窄的防治。
Observe the expression of liposome-mediated vascular endothelial growth factor(VEGF)gene of invitro vascular smooth muscle cells(VSMC), and compare the effect of VEGF on proliferation of vessel endothelium cell andVSMC. METHODS: An eukaryotic expression vector pSV121 containing VEGF cDNA was transferred into VSMC in vitroby lipofectamine reagent- mediated method. Vascular endothelial cells(VEC) were cultured with the above VSMC condi-tioned medium. In order to determine the expression of VEGF mRNA in VSMC and VEGF antigen in VSMC conditionalmedium,and compared the effect of VEGF on proiferahon of VEC and VSMC, Northem blot, Westem blot and [3~H] -thymidine ([~3H]TdR) incpehon were used- RESULTS: The exprssion of VEGF mRNA in trigene VSMC grpiswas higher. The expression of VEGF anhgen in transgenic mediurn was significantly (P < 0.01) than control group,[~3H]TdR data(counts'min) was significantly higher in VEC groups added transgenic medium than control group(P <0.01 ), but there was no significant difference between VSMC groups. CONCLUSION: The methed of liposom-mediatedVEGF gene transfer into VSMC is successful. VEGF might promete proliferation of VEC, but didn't cause proliferationofVSMC, which is favorable for preventing and treating ischendc disease and artery restenosis.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
1999年第10期874-876,共3页
Chinese Journal of Pathophysiology
基金
湖北省重点科技发展项目!96B63
中国博士后基金!1996
关键词
脂质体
平滑肌细胞
血管内皮细胞
VEGF
Endothelial growth factors
Liposomes
Transfection
Muscle, smooth
