溶血磷酯酸诱导白细胞介素13受体α2 mRNA表达并抑制胶原蛋白合成

LPA-induced expression of IL-13Rα2 and inhibitory effect of LPA on synthesis of collagen

目的探讨溶血磷脂酸(lysophosphatidic acid,LPA)对人肺成纤维细胞(HFL-1)白细胞介素-13受体α2(IL-13Rα2)表达和Ⅰ型胶原(collagen typeⅠ,COLAⅠ)合成的影响。方法细胞培养,显微镜观察LPA对HFL-1细胞生长的影响,提取HFL-1细胞总RNA,用RT-PCR检测LPA对HFL-1细胞IL-13Rα2 mRNA和COLA I mRNA表达的影响。用免疫组化方法检测LPA和白细胞介素-13(interleukin-13,IL-13)对HFL-1细胞COLA I蛋白表达的影响。图像分析RT-PCR电泳条带和免疫组化图像,统计学处理。结果①LPA诱导HFL-1细胞IL-13Rα2 mRNA的表达,并呈时间依赖性。②1μmol.L-1 LPA刺激HFL-1细胞,IL-13Rα2 mR-NA的表达增强。③LPA作用HFL-1细胞,对IL-13Rα1的表达无影响。④LPA抑制HFL-1细胞COLA I mRNA的表达,IL-13促进COLA I mRNA的表达,LPA+IL-13混合组COLA I mRNA表达水平比LPA组高,比IL-13组低。⑤免疫组化结果与RT-PCR结果一致。结论溶血磷脂酸诱导人肺成纤维细胞表达IL-13 Rα2 mRNA,并下调人肺成纤维细胞合成COLAⅠ。

Aim To study the effect of lysophosphatidic acid（LPA） on interleukin 13 receptor α2（IL-13Rα2） gene expression and collagen type I（COLA Ⅰ） synthesis in lung fibroblasts.Methods Cell culture was used.The growth of lung fibroblasts HFL-1 cells was observed with microscope.HFL-1 cells were stimulated with LPA in different concentrations.The expression of IL-13Rα2 mRNA and COLA Ⅰ was evaluated by reverse transcriptase polymerase chain reaction（RT-PCR）.The expression of COLA Ⅰ protein was evaluated by immunohistochemisty.Electrophoresis strip and immunohistochemical image were analysed by image analysis.All results were processed by statistics.Results ① LPA enhanced the expression of IL-13Rα2 mRNA in HFL-1 cells in a time-dependent manner.② With the increasing of the concentration of LPA,IL-13 Rα2 mRNA expression significantly increased.After 1 μmol·L-1 LPA stimulated HFL-1 cells,IL-13 Rα2 mRNA expression reached peak.③ LPA had no effect on IL-13Rα1 expression of HFL-1.④ COLA Ⅰ expression of HFL-1 cells was reduced with LPA stimulation.The expression of COLA I mRNA was increased with IL-13.The expression of COLA I mRNA was higher in LPA＋IL-13 group than LPA group and lower than IL-13 group.⑤ Immunohistochemical results and RT-PCR results were consistent.Conclusion LPA induces the expression of IL-13 Rα2 mRNA and downregulates the synthesis of COLA1 in lung fibroblasts.