CAJ | 학술논문

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／mL）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／mL组、1μmol／mL组、3μmol／mL组、10μmol／mL组、30μmol／mL组，每组加入PGD2刺激剂TGF-β2（2．5 ng／mL）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／mL）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。
목적：탐토전렬선소D2（ PGD2）대소서폐성섬유세포TGF-β1／Smads신호통로적영향，위효천기도중소제공분자연구기출。방법채용수궤분조적방법，분별채용불동농도적PGD2수체억제제Laropiprant（0．3、1、3、10、30μmol／mL）불동시간（12、24、48、72、96 h）작용우폐성섬유세포，채용MTT법검측Laropiprant대우세포생장적억제작용。설정상대조조、Laropiprant 0．3μmol／mL조、1μmol／mL조、3μmol／mL조、10μmol／mL조、30μmol／mL조，매조가입PGD2자격제TGF-β2（2．5 ng／mL）배양24 h후，재가입상응농도적Laropiprant자격24 h，분별용PCR법화Western blotting법검측세포TGF-β1、Smad3이급Smad4적표체。결과가입TGF-β2（2．5 ng／mL）처리24 h후，수착Laropiprant적농도증가，세포TGF-β1、Smad3이급Smad4적mRNA급단백표체여정상대조조상비정하강추세（P균＜0．05）。불동농도적Laropiprant작용우세포불동시간후，세포생장억제솔수Laropiprant농도증고화작용시간연장정상승추세，Laropiprant재농도체도1μmol／L，작용시간위24～96 h시，세포생장억제솔명현제고。결론L-929소서폐성섬유세포중PGD2가능통과조절TGF-β1／Smads신호통로인기기도중구。
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 30μmol/mL) at differ-ent 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 0.3μmol/mL group, 1μmol/mL group, 3μmol/mL group, 10μmol/mL group and 30μmol/mL group.Each group was added PGD2 stimulant TGF-β2 (2.5 ng/mL) 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/mL) 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 pro-longed culture time.When Laropiprant concentration reached 1μmol/L, and the reaction time was 24 to 36 h, the cell growth in-hibition rate significantly improved.Conclusion PGD2 may cause the airway remodeling by regulating TGF-β1/Smads signaling pathway in L-929 mouse lung fibroblasts.