Title

tPA和VEGF165基因共表達質粒對人血管細胞增殖的影響和纖溶作用

Translated Titles

Effects of co-expression Plasmid of Human tPA and VEGF165 on the Proliferation of Vascular Cells and Fibrinolysis Activity

Authors

吳忠均(Zhong-Jun Wu);時德(De Shi);駱旭東(Xu-Dong Luo);鄭樹森(Shu-Sen Zheng)

Key Words

組織纖溶酶原激活物 ; 血管內皮生長因子 ; 共表達質粒 ; 血管內皮細胞 ; 血管平滑肌細胞 ; tissue-plasminogen activator ; vascular endothelia growth factor ; co-expression plasmid ; vascular endothelia cell ; vascular smooth muscle cell

PublicationName

中南大學學報(醫學版)

Volume or Term/Year and Month of Publication

30卷4期(2005 / 08 / 01)

Page #

379 - 383

Content Language

簡體中文

Chinese Abstract

目的:觀察組織纖溶酶原激活物(tPA)和血管內皮細胞生長因子l65(VEGF165)基因共表達質粒在血管平滑肌細胞(VSMC)中的表達,並研究表達產物對血管內皮細胞(VEC)和VSMC增殖的影響和纖溶作用。方法:用脂質體轉染法將tPA和VEGF165的共表達質粒pBudCE4.1/tPA-VEGF165轉染VSMC,逆轉錄-聚合酶鏈式反應(RT-PCR)和酶聯免疫吸附實驗(ELISA)分別從mRNA水平和蛋白質水平檢測tPA和VECF165的表達;纖溶蛋白板法檢測轉染基因的VSMC培養基中tPA的纖溶活性;取轉染pBudCE4.1/tPA-VEGF165質粒的VSMC培養基培養VEC和VSMC,用四甲基偶氮唑鹽(MTT法)和流式細胞技術檢測轉基因VSMC的細胞培養基對VEC和VSMC增殖的影響。結果:pBudCE4.1/Tpa-VEGF165轉染VSMC後,RT-PCR和ELISA檢測發現,tPA和VEGF165在mRNA和蛋白質水平均有表達;轉基因培養基有明顯促進纖溶和促VEC增殖作用,而對VSMC增殖無作用。結論:tPA和VEGF165基因共表達質粒pBudCE4.1/tPA-VEGF165能在VSMC表達有生物學活性的tPA和VEGF165,為tPA和VEGF165基因轉染防治移植心臟內血管狹窄奠定了基礎。

English Abstract

Objective To observe the expression of the co-expression plasmid of tissue-plasminogen activator (tPA) and vascular endothelia growth factor 165 (VEGF165) in vascular smooth muscle cells (VSMC) and to study the effect of expressing products on the proliferation of VEC and VSMC and fibrinolysis activity. Methods The co-expression plasmid of tPA and VEGF165 (pBudCE4.1/tPA-VEGF165) was transfected into VSMC with the lipofection. The expression of tPA and VEGF165 at mRNA level was detected by RT-PCR and the protein level expression was detected by enzyme linked immunosorbent assay (ELISA). The fibrinolysis activity of culture medium of VSMC transfected tPA and VEGF165 genes was detected by fibrin plate technique. The VEC and VSMC were cultured with culture medium of VSMC transfected tPA and VEGF165 genes. And the proliferation of VEC and VSMC was evaluated with monoterazolium (MTT) and flow cytometry (FCM). Results The expression of PA and VEGF165 at mRNA and protein levels in the transfected VSMC was demonstrated by RT-PCR and ELISA, respectively. The VSMC culture medium of transfected genes possessed evidently fibrinolysis activity. The expression products of tPA and VEGF165 in the VSMC had an evident effect on the VEC proliferation. But it had not an effect on the VSMC proliferation. Conclusion The eukaryotic co-expression plasmid of tPA and VEGF165 can be expressed in transfected VSMCs. The expression products have an obvious biological activity. The present study lay the foundation for future making use of tPA and VEGF165 to prevent and treat vascular stenosis in transplanted heart.

Topic Category 醫藥衛生 > 醫藥衛生綜合
醫藥衛生 > 預防保健與衛生學
醫藥衛生 > 中醫藥學
醫藥衛生 > 藥理醫學
醫藥衛生 > 社會醫學