组织因子途经抑制物基因对血管内皮细胞分泌层黏连蛋白的影响

Effect of tissue factor pathway inhibitor gene on secretion of laminin in vascular endothelial cells

目的研究组织因子途经抑制物(TFPI)基因转移对大鼠血管内皮细胞层黏连蛋白(LN)表达的影响,探讨TFPI基因抑制再狭窄的机制。方法用丙酮酸钠刺激大鼠血管内皮细胞,比较丙酮酸钠刺激前后大鼠血管内皮细胞表达LN的差异;分别使用携带TFPI基因(Ad-TFPI)和携带LacZ基因(Ad-LacZ)的重组腺病毒在体外转染大鼠血管内皮细胞;用X-gal染色法检测外源基因的转染效率,用酶联免疫吸附测定方法检测TFPI基因转染与LAZ基因转染,大鼠血管内皮细胞分泌LN的浓度变化。结果丙酮酸钠可明显刺激大鼠血管内皮细胞分泌LN。TFPI基因转染培养的大鼠血管内皮细胞后,第2天上清中LN表达减少,为(230.73±32.57)μg/L,Ad-LacZ组为(322.87±34.87)μg/L,两组比较差异有统计学意义(P<0.05),但是第4天和第6天LN表达,TFPI组与Ad-LacZ组比较差异无统计学意义(P>0.05)。结论 TFPI基因转移可明显抑制大鼠血管内皮细胞分泌LN,随着时间的推移这种抑制作用减弱;TFPI可能通过直接抑制LN的表达抑制再狭窄的发生和发展。

Objective The study was to investigate the effect of the transference of tissue factor pathway inhibitor（TFPI）gene on the expression of laminin into the rat vascular endothelial cells and further explore the mechanism that TFPI inhibits restenosis.Methods We stimulated rat vascular endothelial cells to express laminin with sodium pyruvate.The concentration of laminin was compared before and after stimulation by the sodium pyruvate.After transferring adenoviral-mediated TFPI gene and LacZ gene into cultured rat vascular endothelial cell respectively,the transference rate was tested by X-gal staining and the expression of laminin with or without the transference of TFPI gene was detected by enzyme-linked immunosorbent assay.Results Sodium pyruvate stimulated rat vascular endothelial cells to secrete laminin obviously.The expression of laminin after the transference of TFPI into the rat vascular endothelial cells was reduced significantly as compared with that of LacZ,（230.73±32.57）μg/L vs（322.87±34.87）μg/L（P 0.05）,and it was insignificant in the fourth day,and the sixth day between two groups（P 0.05）.Conclusion The data indicate that the inhibition happens mainly in the early stage and diminishes with time.TFPI inhibits restenosis by diminishing the expression of laminin in the early stage.