目的：研究IL－32γ可否诱导肝星状细胞LX－2表达Ⅰ型胶原（ Collagen Ⅰ）。方法不同浓度的IL－32γ与LX－2共培养，分别收集培养上清液、提取细胞总RNA，real－time PCR检测Collagen Ⅰ mRNA表达水平，ELISA法检测CollagenⅠ蛋白质表达水平。用不同浓度的人重组磷酸化p38α蛋白（ recombinant human active p38α蛋白）与LX－2细胞共培养，48 h后收集细胞培养上清液，ELISA法检测Collagen Ⅰ蛋白质表达水平。再用不同浓度的p38MAPK抑制剂SB203580加入LX－2细胞与IL－32γ共培养的培养液中，分别收集培养上清液，采用ELISA检测CollagenⅠ蛋白质表达水平，同时提取细胞总RNA采用real－time PCR检测Collagen Ⅰ mRNA表达水平。结果 IL－32γ诱导人LX－2细胞表达CollagenⅠ，且其表达水平随IL－32γ浓度的增加而增强；人重组磷酸化p38α蛋白可诱导LX－2细胞表达Collagen Ⅰ增高，阻断p38MAPK可降低IL－32γ诱导的Collagen Ⅰ表达。结论 IL－32γ通过活化p38MAPK通路诱导LX－2细胞表达CollagenⅠ，IL－32γ可能通过诱导肝星状细胞表达Collagen Ⅰ而参与肝纤维化的形成。%Objective To investigate whether could IL-32γinduce Collagen Ⅰ expression in LX-2 cells. Methods LX-2 cells were co-cultured with IL-32γof different concentrations.Then total RNA was extracted, and the cell supernatant was collected.CollagenⅠRNA expression level was assayed by using real-time PCR.CollagenⅠprotein expression level was detected by using ELISA.Subsequently, LX-2 cells were co-cultured with recombinant Human Active p38αprotein of different concentrations.Forty-eight hours later, the cell supernatant was collected and CollagenⅠprotein expression level was assessed.The LX-2 cells were then co-cultured with IL-32γ, and p38MAPK inhibitors SB 203580 of different concentrations were added.CollagenⅠRNA and protein expression levels were assessed by using real-time PCR and ELISA, respectively.Results IL-32γcould induce Collagen Ⅰ expression in LX-2 cells.Moreover, CollagenⅠexpression was induced by IL-32γand recombinant Human Active p38αprotein at dose-dependent manners.CollagenⅠexpression induced by IL-32γwas inhibited by p38MAPK.Conclusion IL-32γin-duces CollagenⅠexpression in LX-2 cells via the activation of p38MAPK signal pathway.IL-32 might be involved in the pathogenesis of liver fibrosis via the induction of CollagenⅠexpression in LX-2 cells.
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