重庆医学
重慶醫學
중경의학
CHONGQING MEDICAL JOURNAL
2014年
27期
3588-3591
,共4页
前列腺素D2%TGF-β1%Smads
前列腺素D2%TGF-β1%Smads
전렬선소D2%TGF-β1%Smads
PGD2%transforming growth factor β1%Smads
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。
目的:探討前列腺素D2(PGD2)對L-929小鼠肺成纖維細胞生物學特性的影響,利用其受體特異性抑製劑Laropi-prant調控TGF-β1/Smads信號通路。方法按Laropiprant濃度梯度將細胞分為0.3μmoL組、1.0μmoL組、3.0μmoL組、10.0μmoL組、30.0μmoL ,每組分彆加入TGF-β2(2.5 ng/mL)培養24 h後使用相應濃度的Laropiprant刺激24 h ,併分彆用PCR法和Western blot法檢測細胞TGF-β1、Smad3以及Smad4的錶達。分彆採用不同濃度的Laropiprant用于細胞不同時間(12、24、48、72 h),通過MTT法檢測 Laropiprant對細胞生長的抑製作用。結果隨著 Laropiprant 的濃度增加,細胞 TGF-β1、Smad3以及Smad4的mRNA和蛋白的錶達呈下降趨勢,與對照組比較差異有統計學意義(P<0.05)。不同濃度的Laropiprant作用于細胞不同時間後,細胞抑製率隨濃度增高和作用時間增長呈下降趨勢。結論 L-929小鼠肺成纖維細胞PGD2-DP1的錶達可能與 TGF-β1/Smads的調節相關。Laropiprant作用于細胞後,細胞抑製率隨濃度增高和作用時間增長呈下降趨勢。
목적:탐토전렬선소D2(PGD2)대L-929소서폐성섬유세포생물학특성적영향,이용기수체특이성억제제Laropi-prant조공TGF-β1/Smads신호통로。방법안Laropiprant농도제도장세포분위0.3μmoL조、1.0μmoL조、3.0μmoL조、10.0μmoL조、30.0μmoL ,매조분별가입TGF-β2(2.5 ng/mL)배양24 h후사용상응농도적Laropiprant자격24 h ,병분별용PCR법화Western blot법검측세포TGF-β1、Smad3이급Smad4적표체。분별채용불동농도적Laropiprant용우세포불동시간(12、24、48、72 h),통과MTT법검측 Laropiprant대세포생장적억제작용。결과수착 Laropiprant 적농도증가,세포 TGF-β1、Smad3이급Smad4적mRNA화단백적표체정하강추세,여대조조비교차이유통계학의의(P<0.05)。불동농도적Laropiprant작용우세포불동시간후,세포억제솔수농도증고화작용시간증장정하강추세。결론 L-929소서폐성섬유세포PGD2-DP1적표체가능여 TGF-β1/Smads적조절상관。Laropiprant작용우세포후,세포억제솔수농도증고화작용시간증장정하강추세。
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 .