经颈静脉肝内门腔静脉分流术中胆汁漏出对内皮细胞生长及功能的影响

Bile leakage in transjugular intrahepatic portosystemic shunt: effect on growth and function of endothelium

目的 探讨胆汁对离体培养的内皮细胞生长及功能的影响 ,进一步了解胆汁对支架内皮化的影响。方法 取人脐静脉内皮细胞进行体外培养 ,分别加入 5 %、10 %、15 %、2 0 %、2 5 %胆汁干预 ,观察内皮细胞生长状况 ,收获的细胞测总蛋白 ,四唑盐 (MTT)吸光度值 ,条件培养液测血管性假性血友病因子 (vonWillebrandfactor,简称vWF)行内皮细胞鉴定及功能测定。结果 含 5 %、10 %、15 %胆汁的细胞生长状况与不含胆汁者相似 ,含 2 0 %、2 5 %胆汁的细胞明显减少并显幼稚 ;含 2 5 %胆汁的细胞MTT吸光度值及总蛋白较无胆汁者降低 ,差异有非常显著性意义 (Kruskal Wallis秩和检验 ,χ2 =2 9 913,P =0 0 0 0 9及 χ2 =18 85 7,P =0 0 0 2 ) ;含 2 0 %以上浓度胆汁的细胞条件培养液中vWF含量也明显降低 ,差异有非常显著性意义 (Kruskal Wallis秩和检验 ,χ2 =2 7 2 13,P =0 0 0 0 1)。所收获的细胞vWF测定均阳性。结论 一定浓度的胆汁具有抑制内皮细胞的生长及分泌vWF的功能。所收获的细胞vWF测定均阳性。

Objective To evaluate the effect of bile on growth and function of endothelial cells in vitro. Methods In vitro, endothelial cells explanted from human umbilical vein were cultured. The experiment was designed as 6 groups with concentration of 5%, 10%, 15%, 20%, 25% bile and without bile respectively. Morphological changes of the cells was evaluated daily until to harvest after culture. MTT value of light absorption and total protein was assessed at 5 days. von Willebrand factor (vWF) was measured for identification of cells and estimation of function. Results The explanted cells were verified as endothelial cells by vWF test. Viability and morphology of endothelial cells in culture mediums added with 5%, 10%, 15% bile were similar to those cells without bile, while cells in mediums with 20% and 25% bile were markedly reduced and immature. Comparing with the cells cultured without bile, MTT value of light absorption and total protein in mediums with 25% bile reduced statistical significantly ( P <0 01), while cells in mediums with 5%, 10%, 15% bile were not. The vWF secretion of the cells in mediums with 20% and 25% bile reduced statistical significantly ( P <0.01) compared with the group without bile. Conclusion Certain concentration of bile appears to inhibit endothelial growth and vWF secretion.