中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
4期
805-807
,共3页
刘晗%王玲莉%程剑剑%王存真
劉晗%王玲莉%程劍劍%王存真
류함%왕령리%정검검%왕존진
γ-分泌酶抑制剂%转化生长因子-1%上皮细胞间充质转分化%Notch信号通路
γ-分泌酶抑製劑%轉化生長因子-1%上皮細胞間充質轉分化%Notch信號通路
γ-분비매억제제%전화생장인자-1%상피세포간충질전분화%Notch신호통로
γ-Secretase Inhibitor%Transforming growth factor-β1%Epithelial-to-mesenchymal transition%Notch signaling pathway
目的 探讨γ-分泌酶抑制剂(DAPT)对大鼠肾小管上皮(NRK52E)细胞间充质转分化(EMT)的抑制作用及机制.方法 含10μg/L转化生长因子-β1(TGF-β1)的无血清培养基孵育NRK52E细胞48 h构建EMT模型;或10 mol/L DAPT预处理15 min后构建NRK52E细胞EMT模型.Western blot法检测NRK52E细胞Notch-1、Hes-1、α-平滑肌肌动蛋白(α-SMA)、E-钙黏素(E-cadherin)蛋白表达,小室迁移实验检测细胞迁移能力.结果 TGF-β1显著上调NRK52E细胞Notch-1、Hes-1、α-SMA蛋白表达,下调E-cadherin表达,并增强细胞迁移能力[(482.87±78.23)个比(1464.16± 31.82)个,P＜0.05].DAPT可显著抑制TGF-β1诱导的NRK52E细胞Notch-1、Hes-1、E-cadherin、α-SMA蛋白表达的变化及迁移能力的增强[(509.23 ±83.21)个比(1493.64±463.34)个、(983.06±169.49)个,P＜0.05].结论 Notch信号通路的活化参与了TGF-β1诱导NRK52E细胞EMT,DAPT阻断Notch信号通路活性,抑制TGF-β1诱导的NRK52E细胞EMT.
目的 探討γ-分泌酶抑製劑(DAPT)對大鼠腎小管上皮(NRK52E)細胞間充質轉分化(EMT)的抑製作用及機製.方法 含10μg/L轉化生長因子-β1(TGF-β1)的無血清培養基孵育NRK52E細胞48 h構建EMT模型;或10 mol/L DAPT預處理15 min後構建NRK52E細胞EMT模型.Western blot法檢測NRK52E細胞Notch-1、Hes-1、α-平滑肌肌動蛋白(α-SMA)、E-鈣黏素(E-cadherin)蛋白錶達,小室遷移實驗檢測細胞遷移能力.結果 TGF-β1顯著上調NRK52E細胞Notch-1、Hes-1、α-SMA蛋白錶達,下調E-cadherin錶達,併增彊細胞遷移能力[(482.87±78.23)箇比(1464.16± 31.82)箇,P＜0.05].DAPT可顯著抑製TGF-β1誘導的NRK52E細胞Notch-1、Hes-1、E-cadherin、α-SMA蛋白錶達的變化及遷移能力的增彊[(509.23 ±83.21)箇比(1493.64±463.34)箇、(983.06±169.49)箇,P＜0.05].結論 Notch信號通路的活化參與瞭TGF-β1誘導NRK52E細胞EMT,DAPT阻斷Notch信號通路活性,抑製TGF-β1誘導的NRK52E細胞EMT.
목적 탐토γ-분비매억제제(DAPT)대대서신소관상피(NRK52E)세포간충질전분화(EMT)적억제작용급궤제.방법 함10μg/L전화생장인자-β1(TGF-β1)적무혈청배양기부육NRK52E세포48 h구건EMT모형;혹10 mol/L DAPT예처리15 min후구건NRK52E세포EMT모형.Western blot법검측NRK52E세포Notch-1、Hes-1、α-평활기기동단백(α-SMA)、E-개점소(E-cadherin)단백표체,소실천이실험검측세포천이능력.결과 TGF-β1현저상조NRK52E세포Notch-1、Hes-1、α-SMA단백표체,하조E-cadherin표체,병증강세포천이능력[(482.87±78.23)개비(1464.16± 31.82)개,P＜0.05].DAPT가현저억제TGF-β1유도적NRK52E세포Notch-1、Hes-1、E-cadherin、α-SMA단백표체적변화급천이능력적증강[(509.23 ±83.21)개비(1493.64±463.34)개、(983.06±169.49)개,P＜0.05].결론 Notch신호통로적활화삼여료TGF-β1유도NRK52E세포EMT,DAPT조단Notch신호통로활성,억제TGF-β1유도적NRK52E세포EMT.
Objective To explore the protective effect of γ-secretase inhibitor (DAPT) on epithelial-to-mesenchymal transition (EMT) in NRK 52E cells and the potential mechanism.Methods NRK52E cells were treated with 10 μg/L of transforming growth factor-β1 (TGF-β1) or dimethylsurfoxide (DMSO) for48 h,or NRK52E cells preincubated with 10 μmol/L DAPT for 15 min before 10 μ g/L TGF-β1 treatment for 48 h.Western blotting was applied to detect the protein expression of Notch-1,Hes-1,α-smooth muscle actin (α-SMA) and E-cadherin of NRK52E cells,and transwell assay was applied to measure cell migration.Results TGF-β1 upregulated Notch-1,Hes-1 and α-SMA expression,downregulated E-cadherin expression,and enhanced migration (482.87 ±78.23 vs.1 464.16 ±31.82,P ＜O.05) in NRK52E cells.DAPT treatment attenuated the degree of Notch-1,Hes-1 and α-SMA upregulation expression,E-cadherin suppression and migration (509.23 ± 83.21,1 493.64 ± 463.34 vs.983.06 ± 169.49,P ＜ 0.05 for all) induced by TGF-β1 in NRK52E cells.Conclusion The Notch signaling pathway activation played an important role in TGF-β1-induced EMT in NRK52E cells.DAPT inhibited Notch signaling pathway and protected NRK52E cells from TGF-β1-induced EMT.
