地塞米松抑制胆管瘢痕成纤维细胞的体外研究

Inhibitory effect of Dexamethasone on rabbit bile duct scar fibroblasts in vitro

摘要:

研究地塞米松（DEX）对体外培养的兔胆管瘢痕模型成纤维细胞（MF）中I 型胶原、结缔组织生长因子（CTGF）及第10 号染色体丢失的张力蛋白同源的磷酸酶基因（ PTEN）表达的影响。方法对家兔采用切开再吻合方法复制胆管瘢痕模型并获取MF，经细胞鉴定后对MF 给予不同浓度的DEX（0.00、0.01、0.05 和0.25 mg/ml）干预48 h，活细胞计数法试剂盒测定各组细胞增殖水平；实时聚合酶链反应检测各组细胞I 型胶原、CTGF和PTEN mRNA表达水平；Western blot检测各组细胞I 型胶原和PTEN 蛋白表达水平。结果在DEX 干预48 h后，MF增殖受抑制，细胞I 型胶原、CTGF mRNA 和蛋白表达下调，PTEN mRNA 和蛋白表达上调（p <0.05），呈浓度依耐性。结论DEX可能具有抑制胆管瘢痕及良性胆道狭窄形成的作用，其机制之一可能与MF 中I 型胶原、CTGF 及PTEN的调控有关。

Abstract:

To investigate the effect of Dexamethasone (DEX) on expressions of collagen I,connective tissue growth factor (CTGF ), and phosphatase and tensin homolog deleted on chromosome 10(PTEN ) genes in rabbit bile duct scar model fibroblasts (MF)in vitro . Methods Rabbit bile duct scar model was established to culture MF. The MF were identified, and then treated by different concentrations of DEX (0, 0.01, 0.05 and 0.25 mg/ml). After respective incubation for 48 h, cell proliferation was investigated by CCK-8; relative mRNA expressions of collagen-I, CTGF and PTEN were assessed by qRT-PCR; relative protein expressions of collagen-I and PTEN were investigated by Western blot. Results DEX significantly inhibited cell proliferation, down-regulated both mRNA and protein expressions of collagen-I and , and significantly up-regulated both mRNA and protein expressions of PTEN (p < 0.05) in the MF in a dosedependent manner. Conclusions DEX may have preventive and therapeutic effect on bile duct scar and benign biliary stricture. Effect of DEX in anti-bile duct scar and benign biliary stricture may be partly through regulating collagen-I, CTGF and PTEN expressions in MF.