中华整形外科杂志
中華整形外科雜誌
중화정형외과잡지
CHINESE JOURNAL OF PLASTIC SURGERY
2012年
1期
39-43
,共5页
纪影畅%李宇%鲁峰%胡志奇%王森%林常敏%高建华
紀影暢%李宇%魯峰%鬍誌奇%王森%林常敏%高建華
기영창%리우%로봉%호지기%왕삼%림상민%고건화
毛囊%形态发生%器官培养技术%RNA干扰
毛囊%形態髮生%器官培養技術%RNA榦擾
모낭%형태발생%기관배양기술%RNA간우
Hair follicle%Morphogenesis%Organ culture techniques%RNA interference
目的 利用小干扰RNA(siRNA)抑制皮肤Wnt10b基因的表达,观察Wnt10b基因沉默能否抑制毛囊的发育并探讨其潜在机制.方法 化学合成siRNA-Wnt10b,将siRNA转染体外培养的胎鼠背部皮肤,荧光定量PCR检测转染后不同时间段皮肤组织Wnt10b和p-连环蛋白(β-catenin) mRNA的表达,Western blot检测转染后72 h皮肤组织的Wnt10b和β-catenin蛋白含量.将转染后72 h的皮肤组织石蜡包埋、切片,HE染色,镜下观察各组毛囊发育情况并做统计学处理.结果 siRNA-Wnt10b转染后的24、48 h,胎鼠背部皮肤Wnt10b mRNA的表达呈不同程度下降；但β-catenin mRNA表达未随Wnt10b mRNA水平的起伏而明显波动；转染后72 h,Wnt10b蛋白和β-catenin的蛋白表达同时减少,新形成毛囊数量也随之减少(t=3.254,P=0.002).结论 在体外器官培养的环境中,siRNA-Wnt10b能够抑制胎鼠背部皮肤组织Wnt10b蛋白的表达,减少新形成毛囊的数量；Wnt10b可能只在蛋白水平上调控细胞β-catenin的表达.
目的 利用小榦擾RNA(siRNA)抑製皮膚Wnt10b基因的錶達,觀察Wnt10b基因沉默能否抑製毛囊的髮育併探討其潛在機製.方法 化學閤成siRNA-Wnt10b,將siRNA轉染體外培養的胎鼠揹部皮膚,熒光定量PCR檢測轉染後不同時間段皮膚組織Wnt10b和p-連環蛋白(β-catenin) mRNA的錶達,Western blot檢測轉染後72 h皮膚組織的Wnt10b和β-catenin蛋白含量.將轉染後72 h的皮膚組織石蠟包埋、切片,HE染色,鏡下觀察各組毛囊髮育情況併做統計學處理.結果 siRNA-Wnt10b轉染後的24、48 h,胎鼠揹部皮膚Wnt10b mRNA的錶達呈不同程度下降；但β-catenin mRNA錶達未隨Wnt10b mRNA水平的起伏而明顯波動；轉染後72 h,Wnt10b蛋白和β-catenin的蛋白錶達同時減少,新形成毛囊數量也隨之減少(t=3.254,P=0.002).結論 在體外器官培養的環境中,siRNA-Wnt10b能夠抑製胎鼠揹部皮膚組織Wnt10b蛋白的錶達,減少新形成毛囊的數量；Wnt10b可能隻在蛋白水平上調控細胞β-catenin的錶達.
목적 이용소간우RNA(siRNA)억제피부Wnt10b기인적표체,관찰Wnt10b기인침묵능부억제모낭적발육병탐토기잠재궤제.방법 화학합성siRNA-Wnt10b,장siRNA전염체외배양적태서배부피부,형광정량PCR검측전염후불동시간단피부조직Wnt10b화p-련배단백(β-catenin) mRNA적표체,Western blot검측전염후72 h피부조직적Wnt10b화β-catenin단백함량.장전염후72 h적피부조직석사포매、절편,HE염색,경하관찰각조모낭발육정황병주통계학처리.결과 siRNA-Wnt10b전염후적24、48 h,태서배부피부Wnt10b mRNA적표체정불동정도하강；단β-catenin mRNA표체미수Wnt10b mRNA수평적기복이명현파동；전염후72 h,Wnt10b단백화β-catenin적단백표체동시감소,신형성모낭수량야수지감소(t=3.254,P=0.002).결론 재체외기관배양적배경중,siRNA-Wnt10b능구억제태서배부피부조직Wnt10b단백적표체,감소신형성모낭적수량；Wnt10b가능지재단백수평상조공세포β-catenin적표체.
Objective To investigate whether the suppression of Wnt10b by siRNA could prevent the development of hair follicle in the cultured rat embryonic skin. Methods siRNA-Wnt10b was synthesized by chemosynthesis method.The dorsal skin of SD rat at embryos were cultured in DMEM in the presence of different percentage of interfering RNA targeting Wnt10b. Wnt10b/β-catenin expression was analyzed by real-time PCR everyday and by Western blot on the third day. The cultured embryonic skin underwent paraffin embedding,section,HE staining on the third day,in which the number of de novo hair follicle was calculated and statistically analyzed. Results Wnt10b gene in the cultured embryonic skin could be knocked down with the siRNA-based method.β-catenin mRNA was not greatly influenced by the downregulation of Wnt10b mRNA.The number of de novo hair follicle placode in cultured embryonic skin decreased,along with the downregulation of Wnt10b and β-catenin proteins expression.Conclusions The downregulation of Wnt10b mRNA and protein by siRNA reduces the number of de novo hair follicle placode in the cultured rat embryonic skin.Wnt10b may control cytoplasm β-catenin concentration at the protein level.
