溶血磷脂酸对人肺成纤维细胞白细胞介素13受体α2 mRNA表达的影响

Effects of lysophosphatidicacid on expression of interleukine-13 receptor alpha 2 mRNA in human lung fibroblasts

背景:白细胞介素13受体α2与白细胞介素13结合后,不介导白细胞介素13功能,对白细胞介素13起负调控作用。有研究表明,溶血磷脂酸能增加人支气管上皮细胞白细胞介素13受体α2的表达,从而抑制白细胞介素13的作用。目的:验证溶血磷脂酸对人肺成纤维细胞白细胞介素13受体α2 mRNA表达的诱导作用。设计、时间及地点:mRNA水平的观察对照实验,于2007-07/2008-11在南昌大学医学院和江西省医学生物高技术重点实验室完成。材料:人肺成纤维细胞HFL-1购于中科院上海生化和细胞生物研究所。方法:培养HFL-1细胞,行时间依赖和剂量依赖实验。时间依赖实验采用1μmol/L溶血磷脂酸刺激细胞,分别于0,6,12,24h收集细胞。剂量依赖实验分别用0,0.1,1,10μmol/L的溶血磷脂酸刺激细胞,12h收集细胞。主要观察指标:显微镜下观察溶血磷脂酸对HFL-1细胞生长的影响,提取HFL-1细胞总RNA,用反转录-聚合酶链反应及图像分析法检测溶血磷脂酸对HFL-1细胞白细胞介素13受体α2 mRNA表达的影响。结果:溶血磷脂酸不影响HFL-1细胞生长。1μmol/L溶血磷脂酸刺激HFL-1细胞,白细胞介素13受体α2 mRNA的表达增高,并呈时间依赖性。随着溶血磷脂酸浓度的增加,白细胞介素13受体α2 mRNA的表达明显增加。1μmol/L溶血磷脂酸刺激HFL-1细胞,白细胞介素13受体α2 mRNA的表达可达高峰。溶血磷脂酸对HFL-1细胞白细胞介素13受体α1的表达无影响。结论:溶血磷脂酸能促进人肺成纤维细胞HFL-1白细胞介素13受体α2 mRNA的表达。

BACKGROUND： Interleukine-13 receptor α2 （IL-13 Rα2） plays a negative regulation role in IL-13 when combined with IL-13. Studies demonstrate that lysophosphatidicacid （LPA） can inhabit IL-13 by increasing the expression of IL-13 R, 2 in human lung fibroblasts. OBJECTIVE： To study the induction of LPA on IL-13R α2 gene expression in lung fibroblasts. DESIGN, TIME AND SETTING： The observation comparison experiment based on mRNA levels was performed at the Medical College of Nanchang University and Key Laboratory of Medical Biology of Jiangxi Province from July 2007 to November 2008. MATERIALS： Human lung fibroblasts （HFL-1） were purchased from Institute of Biochemistry and cell biology, ISBS, CSA. METHODS： HFL-1 was cultured in both dose and time dependent manner. In time-dependent experiment, HFL-1 cells were stimulated with 1 μmol/L LPA, and collected at hours 0, 6, 12 and 24. In dependent experiment, HFL-1 cells were stimulated with LPA at concentrations of 0, 0.1, 1, 10 μmol/L. The cells were collected after 12 hours. MAIN OUTCOME MEASURES： Effect of LPA on cell growth of HFL-1 was observed by microscope. The expression of IL-13R α2 mRNA was evaluated by reverse transcriptase polymerase chain reaction （RT-PCR） and image analysis. RESULTES： LPA could not influence the growth of HFL-1 cells. When HFL-1 cells were stimulated with 1μmol/L LPA, the expression of IL-13R α 2 mRNA was increased in a time-dependent manner. The expression of IL-13R α2 mRNA increased as LPA dose increasing, which reached a peaked level when stimulated with 1 μmol/L LPA. LPA had no influence on the expression of IL-13Rα1. CONCLUSION： LPA can enhance the expression of IL-13Rα2 in human lung fibroblasts.