PGD_2对小鼠肺成纤维细胞TGF-β_1/Smads信号通路的影响

Effect of PGD_2 on TGF-β_1/ Smads signaling pathway in mouse lung fibroblasts

目的探讨前列腺素D2(PGD2)对小鼠肺成纤维细胞TGF-β1/Smads信号通路的影响,为哮喘气道重塑提供分子研究基础。方法采用随机分组的方法,分别采用不同浓度的PGD2受体抑制剂Laropiprant(0.3、1、3、10、30μmol/m L)不同时间(12、24、48、72、96 h)作用于肺成纤维细胞,采用MTT法检测Laropiprant对于细胞生长的抑制作用。设正常对照组、Laropiprant 0.3μmol/m L组、1μmol/m L组、3μmol/m L组、10μmol/m L组、30μmol/m L组,每组加入PGD2刺激剂TGF-β2(2.5 ng/m L)培养24 h后,再加入相应浓度的Laropiprant刺激24 h,分别用PCR法和Western blotting法检测细胞TGF-β1、Smad3以及Smad4的表达。结果加入TGF-β2(2.5 ng/m L)处理24 h后,随着Laropiprant的浓度增加,细胞TGF-β1、Smad3以及Smad4的mRNA及蛋白表达与正常对照组相比呈下降趋势(P均<0.05)。不同浓度的Laropiprant作用于细胞不同时间后,细胞生长抑制率随Laropiprant浓度增高和作用时间延长呈上升趋势,Laropiprant在浓度达到1μmol/L,作用时间为24~96 h时,细胞生长抑制率明显提高。结论L-929小鼠肺成纤维细胞中PGD2可能通过调节TGF-β1/Smads信号通路引起气道重构。

Objective To investigate the effects of prostaglandin D2( PGD2) on TGF-β1/ Smads signaling pathway in mice lung fibroblasts,and to provide a molecular study basis for airway remodeling of asthma. Methods The lung fibroblasts were randomly treated by different concentrations of PGD2 receptor inhibitor Laropiprant( 0. 3,1,3,10,and 30 μmol / m L) at different time( 12 h,24 h,48 h,72 h and 96 h). The inhibitory effect of Laropiprant on the cell growth was detected by MTT assay.The cells were divided into the control group,Laropiprant 0. 3 μmol / m L group,1 μmol / m L group,3 μmol / m L group,10 μmol /m L group and 30 μmol / m L group. Each group was added PGD2 stimulant TGF-β2( 2. 5 ng/m L) and then was cultured for 24 hours. The expression of TGF-β1,Smad3 and Smad4 was detected by PCR and Western blotting,respectively. Results After being treated with TGF-β2( 2.5 ng/m L) for 24 hours,the mRNA and protein expression of TGF-β1,Smad3 and Smad4 showed a decreased tendency with the increased addition of Laropiprant concentration as compared with that of the control group( all P <0. 05). The cell growth inhibition rate showed a increased tendency with the increased Laropiprant concentration and the prolonged culture time. When Laropiprant concentration reached 1μmol / L,and the reaction time was 24 to 36 h,the cell growth inhibition rate significantly improved. Conclusion PGD2 may cause the airway remodeling by regulating TGF-β1/ Smads signaling pathway in L-929 mouse lung fibroblasts.