首页> 中文期刊>重庆医学 >PGD2调控 TGF-β1在慢性阻塞性肺疾病气道重塑中的作用

PGD2调控 TGF-β1在慢性阻塞性肺疾病气道重塑中的作用

     

摘要

目的:探讨前列腺素D2(PGD2)对L-929小鼠肺成纤维细胞生物学特性的影响,利用其受体特异性抑制剂Laropi-prant调控TGF-β1/Smads信号通路。方法按Laropiprant浓度梯度将细胞分为0.3μmoL组、1.0μmoL组、3.0μmoL组、10.0μmoL组、30.0μmoL ,每组分别加入TGF-β2(2.5 ng/mL)培养24 h后使用相应浓度的Laropiprant刺激24 h ,并分别用PCR法和Western blot法检测细胞TGF-β1、Smad3以及Smad4的表达。分别采用不同浓度的Laropiprant用于细胞不同时间(12、24、48、72 h),通过MTT法检测 Laropiprant对细胞生长的抑制作用。结果随着 Laropiprant 的浓度增加,细胞 TGF-β1、Smad3以及Smad4的mRNA和蛋白的表达呈下降趋势,与对照组比较差异有统计学意义(P<0.05)。不同浓度的Laropiprant作用于细胞不同时间后,细胞抑制率随浓度增高和作用时间增长呈下降趋势。结论 L-929小鼠肺成纤维细胞PGD2-DP1的表达可能与 TGF-β1/Smads的调节相关。Laropiprant作用于细胞后,细胞抑制率随浓度增高和作用时间增长呈下降趋势。%Objective To explore PGD2 biological characteristics in L-929 mice lung fibroblasts cell by Laropiprant is a specific in-hibitor of PGD2 receptor regulation of TGF-β1/Smads signaling pathway .Provide further basis research to explore the molecular mechanisms of airway fibrosis .Methods The cells divided by Laropiprant concentration 0 .3 μmoL group ,1 .0 μmoL group ,3 .0μmoL group ,10 .0 μmoL group ,30 .0 μmoL and the control group .Each group were added TGF-β2 (2 .5 ng/mL) were cultured for 24 h after stimulation with the corresponding concentrations of Laropiprant 24 h ,TGF-β1 Smad3 and Smad4 expression were detected by PCR and WB ,respectively .A randomized approach ,using different concentrations of Laropiprant acts on cells at different times (12 ,24 ,48 ,72 h) by MTT assay for cell growth Laropiprant inhibition .Results TGF-β1 ,Smad3 and Smad4 mRNA expression de-creased with the addition of Laropiprant concentration increases ,there was significant difference compared with the control group with (P<0 .05) .TGF-β1 ,Smad3 and Smad4 protein expression decreased with the addition of Laropiprant concentration increases ,there was significant difference compared with the control group with (P< 0 .05) .Cell growth inhibition rate decreased with Laropiprant concentration increasing and cell culture time growth ,Laropiprant in concentration 1μml ,reaction time was 24 to 36 h ,the cell growth inhibition was significantly improved .Conclusion PGD2-DP1 expression in L-929 mouse lung fibroblasts cell may be associated with TGF-β1/Smads regulation .growth inhibition rate decreased with Laropiprant concentration increasing and cell culture time growth ,re-action time prolong ,the cell growth inhibition was significantly improved .

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