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目的探讨溶血磷脂酸(lysophosphatidic acid,LPA)对人肺成纤维细胞(HFL-1)Ⅰ型胶原(procollagen typeⅠ,PCOLⅠ)合成的影响。方法将HFL-1培养后置于6孔板中,随机分为6组:对照组,LPAⅠ、Ⅱ、Ⅲ组(用1μmol.L-1的LPA刺激6、12、24h后收集细胞),IL-13组(用100μg.L-1的IL-13作用48h后收集细胞),LPA+IL-13组(用1μmol.L-1的LPA刺激12h、100μg.L-1的IL-13刺激48h后收集细胞),每组3孔。用RT-PCR方法检测PCOLⅠmRNA的表达;用免疫组化方法检测PCOLⅠ蛋白的表达。结果免疫组化与RT-PCR方法均显示LPA刺激后,PCOLⅠ的表达降低,IL-13刺激后,PCOLⅠ的表达增高,且LPA+IL-13组PCOLⅠ表达比LPA组较高,比IL-13组较低。结论LPA可以下调HFL-1PCOLⅠ的合成。
Objective To investigate the effect of lysophosphatidic acid (LPA) on the synthesis of procollagen type Ⅰ (PCOL Ⅰ) in human lung fibroblasts (HFL-1). Methods HFL-1 cells were cultured in 6-well plates and randomly divided into 6 groups: control group, LPAⅠ, Ⅱ and Ⅲ groups (cells were harvested after 6, 12 and 24 h stimulation with 1 μmol·L-1 LPA), IL -13group (cells collected after 48h with 100μg.L-1 IL-13), LPA + IL-13group (LPA stimulated with1μmol.L-1 for12h, 100μg.L-1 IL- After collecting cells), 3 wells per group. The expression of PCOLⅠmRNA was detected by RT-PCR. The expression of PCOLⅠprotein was detected by immunohistochemistry. Results Both immunohistochemistry and RT-PCR showed that the expression of PCOLⅠ was decreased after LPA stimulation. The expression of PCOLⅠ was increased after IL-13 stimulation, and the expression of PCOLⅠ in LPA + IL-13 group was higher than that in LPA group Group is lower. Conclusion LPA can down-regulate the synthesis of HFL-1PCOLⅠ.