中华整形外科杂志
中華整形外科雜誌
중화정형외과잡지
CHINESE JOURNAL OF PLASTIC SURGERY
2013年
6期
406-412
,共7页
周孝亮%刘德伍%毛远桂%吕静
週孝亮%劉德伍%毛遠桂%呂靜
주효량%류덕오%모원계%려정
粉防己碱%瘢痕%Ⅰ型胶原%Ⅲ型胶原%Smad 3
粉防己堿%瘢痕%Ⅰ型膠原%Ⅲ型膠原%Smad 3
분방기감%반흔%Ⅰ형효원%Ⅲ형효원%Smad 3
Tetrandrine%Cicatrix%Collagen type Ⅰ%Collagen type Ⅲ%Transforming growth factor-beta 1%Smads signal pathway
目的 观察粉防己碱(tetrandrine,Tet)对兔耳瘢痕增生组织Ⅰ、Ⅲ型胶原和TGF-β1基因表达的影响,探讨粉防己碱抑制瘢痕增生的体内机制.方法 建立兔耳瘢痕模型,并分为生理盐水组、泼尼松龙组、低浓度Tet组(1.0 mg/ml)、高浓度Tet组(7.5 mg/ml),按分组给药.动态观察瘢痕形态学变化;HE和Masson染色分别观察成纤维细胞数量和胶原表达与排列;免疫组化观察Ⅰ、Ⅲ型胶原和TGF-β1表达情况;RT-PCR检测Ⅰ、Ⅲ型胶原和TGF-β1及Smad 3 mRNA表达.结果 ①伤后24 d创面完全表皮化,形成突出皮面、质地韧、暗红色增生样组织块;HE、Masson染色示:瘢痕增生样组织块成纤维细胞数量及胶原面密度均明显高于正常皮肤,且胶原排列紊乱呈漩涡和结节状.②形态学观察示,Tet组、泼尼松龙组与生理盐水组比较瘢痕充血程度降低、颜色变淡、质地变软、厚度变薄,其中高浓度Tet组改善最明显.③HE、Masson染色显示:粉防己碱组、泼尼松龙组较盐水组成纤维细胞数量明显减少(P＜0.01),胶原面密度明显降低(P＜0.01),排列稀疏整齐,未见漩涡与结节,其中高浓度组成纤维细胞和胶原面密度降低最明显.④免疫组化示:各组Ⅰ、Ⅲ型胶原及TGF-β1均阳性表达;生理盐水组、泼尼松龙组、低浓度Tet组、高浓度Tet组,Ⅰ、Ⅲ型胶原面密度TGF-β1阳性表达率依次降低(P＜0.01);另外Ⅰ、Ⅲ型胶原面密度比值也呈下降趋势,从高到低依次为生理盐水组＞低浓度Tet组＞高浓度Tet组和泼尼松龙组.⑤RT-PCR检测示:各组瘢痕组织Ⅰ、Ⅲ型胶原和TGF-β1及信号分子Smad 3 mRNA扩增条带亮度有明显差异;与生理盐水组比较,Tet组、泼尼松龙组Ⅰ、Ⅲ型胶原和TGF-β1及Smad 3 mRNA表达减弱(P＜0.01);高浓度Tet组对Ⅰ型胶原和TGF-β1及Smad 3 mRNA的表达减低最明显.结论 Tet能明显抑制兔耳瘢痕增生组织Ⅰ、Ⅲ型胶原和TGF-β1基因表达并抑制Smad 3 mRNA表达,可能是其抑制瘢痕增生的重要机制之一.
目的 觀察粉防己堿(tetrandrine,Tet)對兔耳瘢痕增生組織Ⅰ、Ⅲ型膠原和TGF-β1基因錶達的影響,探討粉防己堿抑製瘢痕增生的體內機製.方法 建立兔耳瘢痕模型,併分為生理鹽水組、潑尼鬆龍組、低濃度Tet組(1.0 mg/ml)、高濃度Tet組(7.5 mg/ml),按分組給藥.動態觀察瘢痕形態學變化;HE和Masson染色分彆觀察成纖維細胞數量和膠原錶達與排列;免疫組化觀察Ⅰ、Ⅲ型膠原和TGF-β1錶達情況;RT-PCR檢測Ⅰ、Ⅲ型膠原和TGF-β1及Smad 3 mRNA錶達.結果 ①傷後24 d創麵完全錶皮化,形成突齣皮麵、質地韌、暗紅色增生樣組織塊;HE、Masson染色示:瘢痕增生樣組織塊成纖維細胞數量及膠原麵密度均明顯高于正常皮膚,且膠原排列紊亂呈漩渦和結節狀.②形態學觀察示,Tet組、潑尼鬆龍組與生理鹽水組比較瘢痕充血程度降低、顏色變淡、質地變軟、厚度變薄,其中高濃度Tet組改善最明顯.③HE、Masson染色顯示:粉防己堿組、潑尼鬆龍組較鹽水組成纖維細胞數量明顯減少(P＜0.01),膠原麵密度明顯降低(P＜0.01),排列稀疏整齊,未見漩渦與結節,其中高濃度組成纖維細胞和膠原麵密度降低最明顯.④免疫組化示:各組Ⅰ、Ⅲ型膠原及TGF-β1均暘性錶達;生理鹽水組、潑尼鬆龍組、低濃度Tet組、高濃度Tet組,Ⅰ、Ⅲ型膠原麵密度TGF-β1暘性錶達率依次降低(P＜0.01);另外Ⅰ、Ⅲ型膠原麵密度比值也呈下降趨勢,從高到低依次為生理鹽水組＞低濃度Tet組＞高濃度Tet組和潑尼鬆龍組.⑤RT-PCR檢測示:各組瘢痕組織Ⅰ、Ⅲ型膠原和TGF-β1及信號分子Smad 3 mRNA擴增條帶亮度有明顯差異;與生理鹽水組比較,Tet組、潑尼鬆龍組Ⅰ、Ⅲ型膠原和TGF-β1及Smad 3 mRNA錶達減弱(P＜0.01);高濃度Tet組對Ⅰ型膠原和TGF-β1及Smad 3 mRNA的錶達減低最明顯.結論 Tet能明顯抑製兔耳瘢痕增生組織Ⅰ、Ⅲ型膠原和TGF-β1基因錶達併抑製Smad 3 mRNA錶達,可能是其抑製瘢痕增生的重要機製之一.
목적 관찰분방기감(tetrandrine,Tet)대토이반흔증생조직Ⅰ、Ⅲ형효원화TGF-β1기인표체적영향,탐토분방기감억제반흔증생적체내궤제.방법 건립토이반흔모형,병분위생리염수조、발니송룡조、저농도Tet조(1.0 mg/ml)、고농도Tet조(7.5 mg/ml),안분조급약.동태관찰반흔형태학변화;HE화Masson염색분별관찰성섬유세포수량화효원표체여배렬;면역조화관찰Ⅰ、Ⅲ형효원화TGF-β1표체정황;RT-PCR검측Ⅰ、Ⅲ형효원화TGF-β1급Smad 3 mRNA표체.결과 ①상후24 d창면완전표피화,형성돌출피면、질지인、암홍색증생양조직괴;HE、Masson염색시:반흔증생양조직괴성섬유세포수량급효원면밀도균명현고우정상피부,차효원배렬문란정선와화결절상.②형태학관찰시,Tet조、발니송룡조여생리염수조비교반흔충혈정도강저、안색변담、질지변연、후도변박,기중고농도Tet조개선최명현.③HE、Masson염색현시:분방기감조、발니송룡조교염수조성섬유세포수량명현감소(P＜0.01),효원면밀도명현강저(P＜0.01),배렬희소정제,미견선와여결절,기중고농도조성섬유세포화효원면밀도강저최명현.④면역조화시:각조Ⅰ、Ⅲ형효원급TGF-β1균양성표체;생리염수조、발니송룡조、저농도Tet조、고농도Tet조,Ⅰ、Ⅲ형효원면밀도TGF-β1양성표체솔의차강저(P＜0.01);령외Ⅰ、Ⅲ형효원면밀도비치야정하강추세,종고도저의차위생리염수조＞저농도Tet조＞고농도Tet조화발니송룡조.⑤RT-PCR검측시:각조반흔조직Ⅰ、Ⅲ형효원화TGF-β1급신호분자Smad 3 mRNA확증조대량도유명현차이;여생리염수조비교,Tet조、발니송룡조Ⅰ、Ⅲ형효원화TGF-β1급Smad 3 mRNA표체감약(P＜0.01);고농도Tet조대Ⅰ형효원화TGF-β1급Smad 3 mRNA적표체감저최명현.결론 Tet능명현억제토이반흔증생조직Ⅰ、Ⅲ형효원화TGF-β1기인표체병억제Smad 3 mRNA표체,가능시기억제반흔증생적중요궤제지일.
Objective To observe the effect of tetrandine on gene expression of collagen type Ⅰ 、collagen type Ⅲ 、transformation growth factor-β1 and to investigate the inhibitory effect of tetrandine on the scar tissue hyperplasia in rabbits' ears.Methods After the scar model was formed on the rabbits' ears,the rabbits were divided into 4 groups to receive intro-lesion injection with saline,or prednisolone(Pre) or tetrandrine in low concentration (L-Tet,1.0 mg/ml) or tetrandrine in high concentration (H-Tet,7.5 mg/ml).The morphological changes of scar tissue were observed.The changes of fibroblasts quantity and collagen expression were observed with HE and Masson staining.Immunohistochemical study was used to observe the expression level of collagen type Ⅰ and collagen type Ⅲ and TGF-β1.Collagen type Ⅰ and collagen type Ⅲ and TGF-β1 and signal factor Smad 3 mRNA were detected with RT-PCR.Results ①24 days after injury,all the wounds healed completely with formation of red,tough and hypertrophic scar.HE and Masson staining showed significant increase of fibroblasts and collagen density with irregularly arrangement.②Compared with that in saline group,the scar in other groups became softer,lighter and thinner,especially in H-Tet group.③HE and Masson staining shows the scar in Tet and Pre groups contained less fibroblasts and lower collagen dentsity with comparatively regular arrangement than that in saline group(P ＜ 0.01),especially in H-Tet group.④According to the immunohistochemical study,the expression of collage type Ⅰ and Ⅲ and TGF-β was positive in all the groups,but the positive rate and the ratio of collagen density Ⅰ to Ⅲ decreased in the order of saline,L-Tet,H-Tet and Pre groups (P ＜ 0.01).⑤PT-PCR detection results showed that the amplification bands brightness of collagen type Ⅰ and Ⅲ and TGF-β1 and signal molecular Smad 3 mRNA in scar tissue were obviously different.Compared with that in saline group,the expression of collagen type Ⅰ and Ⅲ and TGF-β1 and Smad 3 mRNA decreased in Tet and Pre groups (P ＜ 0.01).H-Tet group showed the most obvious reduce in the expression of type Ⅰ collagen and TGF-β1 and Smad 3 mRNA.Conclusions Tetrandine can significantly suppress the expression of collagen type Ⅰ and collagen type Ⅲ and TGF-β1 on hypertrophic scar of rabbit ears,and reduce signal factor Smad 3 mRNA' s expression.It may be one of the important mechanism for its inhibitory effect on scar hyperplasia.
