胆汁酸对人肝内胆管瘢痕成纤维细胞生长的影响

The effects of bile acid on the growth of scar fibroblasts in human intrahepatic bile ducts

目的探讨胆汁酸对人肝内胆管瘢痕成纤维细胞生长的影响。方法原代培养人肝内胆管瘢痕组织成纤维细胞。倒置显微镜下观察细胞生长的形态变化,采用噻唑蓝(MTT)法、细胞计数细胞和划痕实验检测胆汁酸对瘢痕成纤维细胞生长的影响。结果 (1)成功原代培养人肝内胆管瘢痕成纤维细胞。(2)MTT显示在72h,浓度0.5、1.0、2.5、5μmol/L的胆酸(CA)和脱氧胆酸(DCA)促进瘢痕成纤维细胞活力(P<0.05),其他胆汁酸对瘢痕成纤维细胞的生长无明显影响(P>0.05);浓度在10、20、50、100、250、500μmol/L,游离胆汁酸和结合胆汁酸抑制瘢痕成纤维细胞的活力(P<0.05)。同时,单独使用CA(2.5μmol/L)或DCA(5μmol/L)或联合使用CA(2.5μmol/L)和DCA(5μmol/L),与未添加胆汁酸比较,细胞数量、活力明显增加,愈合距离明显缩小(P<0.05);联合使用与单独使用比较,细胞数量、活力和愈合距离差异无统计学意义(P>0.05)。结论游离胆汁酸(CA、DCA)在肝内胆管瘢痕的形成过程中可能扮演重要角色。

Objective To investigate the effects of bile acid on the growth of scar fibroblasts in human intrahepat-ic bile ducts.Methods Primary scar fibroblasts from human intrahepatic bile ducts were co-culture with bile acid.Mor-phological changes were observed under microscope.The growth of scar fibroblast was tested by MTT method, cell count and wound-healing test.Results The primary scar fibroblasts of intrahepatic bile ducts were successfully cultured.De-termined by MTT, significant promotion on the vitality of scar fibroblasts was observed in CA and DCA in the concentra-tions of 0.5, 1.0, 2.5, 5 μmol/L （P〈0.05）, while the other bile acids had no effect on the growth of scar fibroblasts （P〉0.05）.However, in concentration of 10, 20, 50, 100, 250, 500 μmol/L, the free bile acids and conjugated bile acids significantly inhibited the growth of scar fibroblasts （P〈0.05）.The count and vitality of the scar fibroblasts with the CA plus DCA, CA alone or DCA alone were significantly higher than the fibroblasts with CA plus DCA, CA alone and DCA alone （P〉0.05）.Conclusion Cholic acid and deoxycholic acid play important role in the formation of scar in the intrahepatic bile ducts.