摘要
目的探讨内皮抑素（ENDO）-血管内皮细胞抑制因子（VEGI）重组腺病毒对同型半胱氨酸（Hcy）致人血管内皮细胞损伤的拮抗作用。方法应用内皮抑素-血管内皮细胞抑制因子融合蛋白重组腺病毒（Ad-hENDOVEGI151）转染Hcy损伤的人血管内皮细胞ECV304,用免疫印迹法检测受染细胞融合蛋白的表达,用自动生化分析仪检测乳酸脱氢酶（LDH）漏出量,用台盼蓝染色法计算细胞存活率,用ELISA检测肿瘤坏死因子α（TNF-α）、单核细胞趋化蛋白1（MCP-1）的表达。结果制备的重组腺病毒具有较高的转基因效率,Ad-hENDO-VEGI151重组腺病毒能够在ECV304细胞中表达41 kDa大小的融合蛋白,Hcy（0.1~1.0 mmol/L）呈浓度依赖性地增加细胞LDH漏出量和降低细胞存活率,重组腺病毒能够有效地拮抗Hcy对人血管内皮细胞的损伤作用（P〈0.05）。结论内皮抑素-血管内皮细胞抑制因子重组腺病毒对Hcy致人血管内皮细胞损伤有拮抗作用,为治疗高同型半胱氨酸血症等疾病提供新的思路。
Aim To investigate the antagonistic effect of endostatin（ ENDO）-vascular endothelial growth inhibitor（ VEGI） recombinant adenoviruses to the vascular endothelial cell injury caused by homocysteine（ Hcy）.Methods The Ad-hENDO-VEGI151 from the fusion protein of ENDO and VEGI was used to transfect the human ECV304 caused by Hcy. The expression of fusion protein was detected by Western blot,the leakage of lactate dehydrogenase（ LDH） was detected by automatic biochemical analyzer,the cell viability was detected by trypan-blue staining method,and the expression of tumor necrosis factor-α（ TNF-α） and monocyte chemoattractant protein-1（ MCP-1） were detected by ELISA. Results The transgenic efficiency of recombinant adenovirus was relatively high. A fusion protein with molecular weight of 41 kDa was expressed in the Ad-hENDO-VEGI151 recombinant adenovirus of ECV304 cells. Hcy（ 0.1 ~1.0 mmol/L） increased the leakage of LDH and decreased the cell viability in concentration-dependent manner. The recombinant adenovirus effectively inhibited the injury of Hcy on human vascular endothelial cells（ P 〈 0. 05）.Conclusion The ENDO-VEGI recombinant adenoviruses had antagonistic effect on vascular endothelial cell injury caused by Hcy,which might provide an idea for future prospecting.
作者
赵东明
刘格
张荣秋
修敏
王海桃
ZHAO Dong-Ming;LIU Ge;ZHANG Rong-Qiu;XIU Min;WANG Hai-Tao(The Second Care Health Department, Qingdao Municiple Hospital, Qingdao 266071, Shandong, China;Department of Pharmacy, Qingdao Tumor Hospital, Qingdao 266042, Shandong , China)
出处
《中国动脉硬化杂志》
CAS
2018年第4期373-376,共4页
Chinese Journal of Arteriosclerosis
