摘要
目的:探索白细胞介素-13( IL-13)对人肝星状细胞( hepatic stellate cell )纤维化作用的影响及其分子机制。方法采用MTT方法观察IL-13对人肝星状细胞株LX-2增殖的影响;RT-PCR技术检测LX-2细胞Ⅰ型胶原蛋白( collagen typeⅠ, COLⅠ) mRNA表达;ELISA和羟脯氨酸实验定量检测IL-13对LX-2细胞分泌COLⅠ的影响;Western blot技术分析IL-13对LX-2细胞的信号转导子和转录激活子6( STAT6)磷酸化的影响。结果 IL-1310 ng/ml组、20 ng/ml组和50 ng/ml组LX-2细胞增殖显著高于空白对照组(P<0.05),并呈现剂量依赖关系;加入IL-1350 ng/ml显著上调了LX-2细胞COLⅠmRNA和蛋白的表达(P<0.05),低浓度IL-13(5 ng/ml、10 ng/ml、20 ng/ml)对LX-2细胞COLⅠmRNA和蛋白的表达无显著影响(P>0.05);IL-13(50 ng/ml)作用LX-2细胞,60 min组和120 min组磷酸化STAT6蛋白表达显著增强(P<0.05)。结论 IL-13促进人肝星状细胞增殖并上调COLⅠmRNA和蛋白表达,IL-13促进人肝星状细胞的纤维化作用的可能机制是激活STAT6磷酸化。
Objective To investigate the effects of IL-13 on fibrosis in hepatic stellate cells and its molecular mechanism .Methods The effects of IL-13 on the proliferation of hepatic stellate cell line LX-2 were measured by MTT assay .The transcription level of collagen typeⅠ( COLⅠ) in LX-2 cells was detec-ted by RT-PCR.The secretion of COLⅠin LX-2 cells was measured by ELISA assay and hydroxyproline as-say.Western blot assay was used to analyze the effects of IL-13 on the phosphorylation of signal transducer and activator of transcription(STAT6).Results Compared with control group, IL-13 (10 ng/ml, 20 ng/ml and 50 ng/ml ) significantly stimulated the proliferation of LX-2 cells in a dose-dependent manner ( P〈0.05).The expression of collagen typeⅠin LX-2 cells at mRNA and protein levels were significantly up-regulated by IL-13 at a concentration of 50 ng/ml (P〈0.05), but not affected by IL-13 at low concentra-tions (5 ng/ml, 10 ng/ml, and 20 ng/ml) (P〉0.05).The expression of phosphorylated STAT6 protein in LX-2 cells was significantly enhanced upon the stimulation with 50 ng/ml of IL-13 ( P〈0.05 ) for60 min or 120 min.C onclusion IL-13 promoted the proliferation of human hepatic stellate cells and up-regulated the expression of COLⅠat mRNA and protein levels .IL-13 might promote the fibrosis in human hepatic stellate cells through activating STAT 6 phosphorylation .
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2014年第5期364-368,共5页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金项目(30860118)
江西省自然科学基金项目(20122BAB205020)