中华整形外科杂志
中華整形外科雜誌
중화정형외과잡지
CHINESE JOURNAL OF PLASTIC SURGERY
2013年
4期
285-289
,共5页
瘢痕疙瘩%成纤维细胞%5-氮杂-2-脱氧胞苷%TGF-β/smad通路
瘢痕疙瘩%成纖維細胞%5-氮雜-2-脫氧胞苷%TGF-β/smad通路
반흔흘탑%성섬유세포%5-담잡-2-탈양포감%TGF-β/smad통로
Keloid%Fibroblasts%5-aza-2-deoxycytidine%TGF-β/smad signaling pathway
目的 探讨甲基化酶抑制剂5-氮杂-2-脱氧胞苷对瘢痕疙瘩成纤维细胞TGF-β/smad信号通路的影响.方法 收集瘢痕疙瘩及正常皮肤组织各15例,免疫组化检测磷酸化smad2(p-smad2)和磷酸化smad3(p-smad3)在瘢痕疙瘩和正常皮肤中的阳性表达率;将瘢痕疙瘩分为实验组和对照组,实验组以5-氮杂-2-脱氧胞苷(5×10-5mmol/L)干预,对照组用等量的DMEM培养液,采用组织块法培养人瘢痕疙瘩成纤维细胞,流式细胞仪分析5-氮杂-2-脱氧胞苷(5×10-5mol/L)对瘢痕疙瘩成纤维细胞周期及凋亡的影响;Western blot检测各组p-smad2、p-smad3、TGF-β1和smad7的表达变化;细胞免疫荧光染色法观察各组瘢痕疙瘩成纤维细胞内p-smad2和p-smad3蛋白的影响.结果 瘢痕疙瘩组织中p-smad2和p-smad3阳性表达率明显高于正常皮肤组织中p-smad2和p-smad3阳性表达.流式细胞仪显示5-氮杂-2-脱氧胞苷干预瘢痕疙瘩成纤维细胞后,细胞停滞于G0/G1期比例增加并且细胞凋亡率增加,瘢痕疙瘩成纤维细胞中p-smad2和p-smad3蛋白的表达减少,同时,TGF-β1蛋白表达减少,smad7蛋白表达回升.此外,5-氮杂-2-脱氧胞苷抑制smad2和smad3磷酸化及核转移.结论 甲基化酶抑制剂5-氮杂-2-脱氧胞苷可抑制瘢痕疙瘩成纤维细胞的增殖及促进其凋亡,其作用机制可能与抑制TGF-β/smad信号通路有关.
目的 探討甲基化酶抑製劑5-氮雜-2-脫氧胞苷對瘢痕疙瘩成纖維細胞TGF-β/smad信號通路的影響.方法 收集瘢痕疙瘩及正常皮膚組織各15例,免疫組化檢測燐痠化smad2(p-smad2)和燐痠化smad3(p-smad3)在瘢痕疙瘩和正常皮膚中的暘性錶達率;將瘢痕疙瘩分為實驗組和對照組,實驗組以5-氮雜-2-脫氧胞苷(5×10-5mmol/L)榦預,對照組用等量的DMEM培養液,採用組織塊法培養人瘢痕疙瘩成纖維細胞,流式細胞儀分析5-氮雜-2-脫氧胞苷(5×10-5mol/L)對瘢痕疙瘩成纖維細胞週期及凋亡的影響;Western blot檢測各組p-smad2、p-smad3、TGF-β1和smad7的錶達變化;細胞免疫熒光染色法觀察各組瘢痕疙瘩成纖維細胞內p-smad2和p-smad3蛋白的影響.結果 瘢痕疙瘩組織中p-smad2和p-smad3暘性錶達率明顯高于正常皮膚組織中p-smad2和p-smad3暘性錶達.流式細胞儀顯示5-氮雜-2-脫氧胞苷榦預瘢痕疙瘩成纖維細胞後,細胞停滯于G0/G1期比例增加併且細胞凋亡率增加,瘢痕疙瘩成纖維細胞中p-smad2和p-smad3蛋白的錶達減少,同時,TGF-β1蛋白錶達減少,smad7蛋白錶達迴升.此外,5-氮雜-2-脫氧胞苷抑製smad2和smad3燐痠化及覈轉移.結論 甲基化酶抑製劑5-氮雜-2-脫氧胞苷可抑製瘢痕疙瘩成纖維細胞的增殖及促進其凋亡,其作用機製可能與抑製TGF-β/smad信號通路有關.
목적 탐토갑기화매억제제5-담잡-2-탈양포감대반흔흘탑성섬유세포TGF-β/smad신호통로적영향.방법 수집반흔흘탑급정상피부조직각15례,면역조화검측린산화smad2(p-smad2)화린산화smad3(p-smad3)재반흔흘탑화정상피부중적양성표체솔;장반흔흘탑분위실험조화대조조,실험조이5-담잡-2-탈양포감(5×10-5mmol/L)간예,대조조용등량적DMEM배양액,채용조직괴법배양인반흔흘탑성섬유세포,류식세포의분석5-담잡-2-탈양포감(5×10-5mol/L)대반흔흘탑성섬유세포주기급조망적영향;Western blot검측각조p-smad2、p-smad3、TGF-β1화smad7적표체변화;세포면역형광염색법관찰각조반흔흘탑성섬유세포내p-smad2화p-smad3단백적영향.결과 반흔흘탑조직중p-smad2화p-smad3양성표체솔명현고우정상피부조직중p-smad2화p-smad3양성표체.류식세포의현시5-담잡-2-탈양포감간예반흔흘탑성섬유세포후,세포정체우G0/G1기비례증가병차세포조망솔증가,반흔흘탑성섬유세포중p-smad2화p-smad3단백적표체감소,동시,TGF-β1단백표체감소,smad7단백표체회승.차외,5-담잡-2-탈양포감억제smad2화smad3린산화급핵전이.결론 갑기화매억제제5-담잡-2-탈양포감가억제반흔흘탑성섬유세포적증식급촉진기조망,기작용궤제가능여억제TGF-β/smad신호통로유관.
Objective To investigate the effect of 5-aza-2-deoxycytidine on the TGF-β/smad signal transduction pathway in human keloid fibroblasts(KFSs).Methods Firstly,immunohistochemical method was used to detect the positive expression rate of phoshpo-smad2 and phoshpo-smad3 in the specimens of 15 cases of keloid and 15 cases of normal skin.The keloid fibroblasts were cultured in vitro with 5-aza-2-deoxycytidine(experimental group) or with DMEM (control group).The effect of 5-aza-2-deoxycytidine on the cell cycle and apoptosis of fibroblasts was analysed with fiowcytometry (FCM).Transforming growth factor (TGF)-β1,Smad7,phoshpo-smad2 and phoshpo-smad3 were analyzed by Western Blot,and Immunofluorescence.Results It was found that the positive expression of phoshposmad2 and phoshpo-smad3 in keloid were higher than those in normal skin.The FCM showed that the proportion of cells in G0/G1 stage was increased,and so does the proportion of apoptosis cells in keloid fibroblasts intervented by 5-aza-2-deoxycytidine.The expression of TGF-β1,phoshpo-smad2 and phoshposmad3 protein were significantly suppressed while the expression of smad7 protein increased in keloid fibroblasts with 5-aza-2-deoxycytidine.In addition,5-aza-2-deoxycytidine reversed phosphorylation and nuclear translocation of smad2 and smad3.Conclusions 5-aza-2-deoxycytidine,methylaze inhibitors,inhibits cell proliferation and promotes apoptosis of KFSs,which may be associated with the suppression of TGF-β/smad signal pathway.
