JAK-STATs通路在CTGF刺激人增生性瘢痕成纤维细胞增殖分化中的作用

The function of JAK-STATs pathway in proliferation and differentiation of human hypertrophic scar fibroblast induced by connective tissue growth factor

目的:探讨JAK-STATs是否参与CTGF刺激人增生性瘢痕成纤维细胞(human hypertrophic scar fibroblast,hHSF)增殖分化过程。方法:原代培养hHSF,将其分为①对照组:hHSF组;②CTGF刺激hHSF组。取0min、5min、10min、20min、30min、45min、60min、90min八个时相点,采用western-blot方法检测各时相点JAK1、JAK2、JAK3、TYK2、STAT1、STAT2、STAT3、STAT4、STAT5、STAT6蛋白活化(磷酸化)情况。再对筛选出的蛋白用免疫荧光化学染色和凝胶阻滞电泳(EMSA)方法进一步验证分析其表达及活化情况,以筛选出参与CTGF调控hHSF增殖和分化的JAK-STATs信号分子。结果:通过western-blot、免疫荧光化学染色和凝胶阻滞电泳(EMSA)方法筛选出JAK1、STAT1蛋白在CTGF刺激后蛋白活化程度明显增强。结论:JAK1、STAT1可能参与了CTGF调控hHSF增殖过程。这从一定程度上揭示了CTGF调控hHSF增殖分化过程的信号转导机制,从而为抑制瘢痕纤维化提供了新的研究方向,有助于更深入地调控CTGF的促纤维化作用,为hHSF及其它纤维化疾病的研究及防治提供新的思路。

Objective The aim of the study is to investigate whether JAK-STATs pathway paticipated in the process of CTGF-induced proliferation and differentiation of human hypertrophic scar fibroblast （hHSF）. Methods To cultivate hHSF with primary cultivation,then,to divide the cells into two groups： （1）hHSF;（2） hHSF with CTGF, Western-blot was used to detect JAK1, JAK2, JAK3, TYK2, STAT1, STAT2, STAT3, STAT4, STAT5, STAT6 at 0min,5 min,10 min,20 min,30 min,45 min,60 min,90 min. And then,immunofluorescence（IMF）and electrophoretic mobility shift assay（EMSA） were used to verify the boltered signalling moleculars and learn more about its activation. After the special signalling moleculars were choosed,in order to clarify the process. Results In the premise that proliferation and differentiation of group 2 was much higher than that of group 1 （P〈0.05）,results of western-blot.IF.EMSA told us JAK1 and STAT1 paticipated in the process of CTGF-induced proliferation and differentiation of hHSF with great possibility. Conclusion JAK1 and STAT1 may be important in the process of CTGF-induced proliferation of hHSF.The results above revealed the signal transduction mechanism of CTGF-induced proliferation of hHSF, may afford new direction to inhibit scar fibrosis and contraction.