CAJ | 학술논문

背景:目前认为转化生长因子β与瘢痕形成关系最密切,是关键的活性分子,可影响瘢痕形成的各个阶段.抑制转化生长因子β的生物学作用,在理论上可减少瘢痕的形成.目的:探讨反义转化生长因子β1脱氧寡核苷酸对增生性瘢痕动物模型伤口愈合中瘢痕生成的抑制作用,观察使用反义转化生长因子β1的有效给药途径.设计:自身对照,动物实验.单位:解放军兰州军区总医院安宁分院整形科.材料:实验于2002-09/2003-07在兰州医学院解剖实验室完成.选择日本大耳白兔20只.方法:在每只兔耳腹侧面沿长轴避开可见血管,作两个1.0 cm×2.5 cm的长方形全层皮肤缺损创面,创面间隔1.5 cm,深达软骨表面,共80个,建立兔耳腹侧面增生性瘢痕模型.待兔耳创面上皮化后(平均20 d),在每只白兔左耳每个创面的上皮下用微量注射器局部封闭注射反义转化生长因子β1脱氧寡核苷酸5μL(1g/L),为转化生长因子β1组;右耳每个创面下注射生理盐水5 μL,为生理盐水对照组.注药后3,7,11,20,30 d 5个时相点切取瘢痕组织,每个时相点4只.采用苏木精-伊红染色、Masson染色及转化生长因子β1 mRNA、Ⅰ、Ⅲ型胶原mRNA的原位杂交组织化学染色.主要观察指标:苏木精-伊红染色、Masson染色和原位杂交组织化学染色结果.结果:纳入动物20只,均进入结果分析.①苏木精-伊红染色显示各组中的右耳增生性瘢痕内均可见有炎性细胞浸润并有明显的白细胞浸润带,而左耳的增生性瘢痕经反义转化生长因子β1干预后虽也有炎性细胞浸润,但未见白细胞浸润带.②Masson染色见右侧耳的增生性瘢痕从伤后3周起均可见蓝染较深的胶原纤维,至第7周时仍可见蓝染的胶原纤维,外形粗大(宽度约为8～10μm),排列杂乱无章.而左耳经反义转化生长因子β1干预的兔耳增生性瘢痕在伤后3周时,虽然亦可见蓝染的胶原纤维,但到第6,7周时胶原纤维蓝染变浅并且纤细(宽度为3～5 μm),排列整齐有序.③原位杂交显示转化生长因子β1 mRNA、Ⅰ型胶原mRNA、Ⅲ型胶原mRNA阳性细胞表达率明显降低.结论:反义转化生长因子β1能抑制兔耳增生性瘢痕的增殖过程,使瘢痕组织纤维化程度明显减轻.局部注射裸DNA治疗瘢痕的给药途径是可行的. 揹景:目前認為轉化生長因子β與瘢痕形成關繫最密切,是關鍵的活性分子,可影響瘢痕形成的各箇階段.抑製轉化生長因子β的生物學作用,在理論上可減少瘢痕的形成.目的:探討反義轉化生長因子β1脫氧寡覈苷痠對增生性瘢痕動物模型傷口愈閤中瘢痕生成的抑製作用,觀察使用反義轉化生長因子β1的有效給藥途徑.設計:自身對照,動物實驗.單位:解放軍蘭州軍區總醫院安寧分院整形科.材料:實驗于2002-09/2003-07在蘭州醫學院解剖實驗室完成.選擇日本大耳白兔20隻.方法:在每隻兔耳腹側麵沿長軸避開可見血管,作兩箇1.0 cm×2.5 cm的長方形全層皮膚缺損創麵,創麵間隔1.5 cm,深達軟骨錶麵,共80箇,建立兔耳腹側麵增生性瘢痕模型.待兔耳創麵上皮化後(平均20 d),在每隻白兔左耳每箇創麵的上皮下用微量註射器跼部封閉註射反義轉化生長因子β1脫氧寡覈苷痠5μL(1g/L),為轉化生長因子β1組;右耳每箇創麵下註射生理鹽水5 μL,為生理鹽水對照組.註藥後3,7,11,20,30 d 5箇時相點切取瘢痕組織,每箇時相點4隻.採用囌木精-伊紅染色、Masson染色及轉化生長因子β1 mRNA、Ⅰ、Ⅲ型膠原mRNA的原位雜交組織化學染色.主要觀察指標:囌木精-伊紅染色、Masson染色和原位雜交組織化學染色結果.結果:納入動物20隻,均進入結果分析.①囌木精-伊紅染色顯示各組中的右耳增生性瘢痕內均可見有炎性細胞浸潤併有明顯的白細胞浸潤帶,而左耳的增生性瘢痕經反義轉化生長因子β1榦預後雖也有炎性細胞浸潤,但未見白細胞浸潤帶.②Masson染色見右側耳的增生性瘢痕從傷後3週起均可見藍染較深的膠原纖維,至第7週時仍可見藍染的膠原纖維,外形粗大(寬度約為8～10μm),排列雜亂無章.而左耳經反義轉化生長因子β1榦預的兔耳增生性瘢痕在傷後3週時,雖然亦可見藍染的膠原纖維,但到第6,7週時膠原纖維藍染變淺併且纖細(寬度為3～5 μm),排列整齊有序.③原位雜交顯示轉化生長因子β1 mRNA、Ⅰ型膠原mRNA、Ⅲ型膠原mRNA暘性細胞錶達率明顯降低.結論:反義轉化生長因子β1能抑製兔耳增生性瘢痕的增殖過程,使瘢痕組織纖維化程度明顯減輕.跼部註射裸DNA治療瘢痕的給藥途徑是可行的.
배경:목전인위전화생장인자β여반흔형성관계최밀절,시관건적활성분자,가영향반흔형성적각개계단.억제전화생장인자β적생물학작용,재이론상가감소반흔적형성.목적:탐토반의전화생장인자β1탈양과핵감산대증생성반흔동물모형상구유합중반흔생성적억제작용,관찰사용반의전화생장인자β1적유효급약도경.설계:자신대조,동물실험.단위:해방군란주군구총의원안저분원정형과.재료:실험우2002-09/2003-07재란주의학원해부실험실완성.선택일본대이백토20지.방법:재매지토이복측면연장축피개가견혈관,작량개1.0 cm×2.5 cm적장방형전층피부결손창면,창면간격1.5 cm,심체연골표면,공80개,건립토이복측면증생성반흔모형.대토이창면상피화후(평균20 d),재매지백토좌이매개창면적상피하용미량주사기국부봉폐주사반의전화생장인자β1탈양과핵감산5μL(1g/L),위전화생장인자β1조;우이매개창면하주사생리염수5 μL,위생리염수대조조.주약후3,7,11,20,30 d 5개시상점절취반흔조직,매개시상점4지.채용소목정-이홍염색、Masson염색급전화생장인자β1 mRNA、Ⅰ、Ⅲ형효원mRNA적원위잡교조직화학염색.주요관찰지표:소목정-이홍염색、Masson염색화원위잡교조직화학염색결과.결과:납입동물20지,균진입결과분석.①소목정-이홍염색현시각조중적우이증생성반흔내균가견유염성세포침윤병유명현적백세포침윤대,이좌이적증생성반흔경반의전화생장인자β1간예후수야유염성세포침윤,단미견백세포침윤대.②Masson염색견우측이적증생성반흔종상후3주기균가견람염교심적효원섬유,지제7주시잉가견람염적효원섬유,외형조대(관도약위8～10μm),배렬잡란무장.이좌이경반의전화생장인자β1간예적토이증생성반흔재상후3주시,수연역가견람염적효원섬유,단도제6,7주시효원섬유람염변천병차섬세(관도위3～5 μm),배렬정제유서.③원위잡교현시전화생장인자β1 mRNA、Ⅰ형효원mRNA、Ⅲ형효원mRNA양성세포표체솔명현강저.결론:반의전화생장인자β1능억제토이증생성반흔적증식과정,사반흔조직섬유화정도명현감경.국부주사라DNA치료반흔적급약도경시가행적.
BACKGROUND: Nowadays, it is thought that transforming growth factorβ (TGFβ) is closely related with cicatrization. TGFβ that is a key active molecule can affect each phase of cicatrization. Theoretically, to inhibit the biological effect of TGF β can reduce cicatrization.OBJECTIVE: To explore the inhibitive effect of antisense TGF β1 deoxy-oligonucleotide on generation of cicatricle in intention of animal models with hyperplastic scars and observe the effective route of administration of using antisense TGF β1.DESIGN: Own control and animal study.SETTING: Department of Plastic Surgery, Anning Hospital of General Hospital, Lanzhou Military Area Command of Chinese PLAMATERIALS: The experiment was performed at the laboratory of anatomy,Lanzhou Medical College from September 2002 to July 2003. Totally 20flap-eared Japanese rabbits were selected.METHODS: Blood vessels could be seen in ventral surface of each rabbits' ear getting out of the way along long axis to establish two 1.0 cm×2.5 cm oblong full-thickness cutaneous deficiency raw surfaces that interval for 1.5 cm, to the surface of cartilage, totally 80, so asto establish ventral surface of rabbits' ear models with hyperplastic scars. After epithelizatio of raw surfaces of rabbits' ear (20 days, averagely), 5μL(1 g/L) antisense TGF β1 deoxy-oligonucleotide was closely injected into local endepidermis of each raw surface of left ear of each rabbit with microinjector, which was regarded as TGF β1 group. 5 μL saline was injected into each raw surface of right ears, which was regarded as saline control group. After injection for3, 7, 11, 20, 30 days, cicatricial tissues were cut, 4 rabbits in each time phase. Hematoxylin-esoin (HE) staining, Masson staining and TGFβ1 mRNA, type Ⅰ and Ⅲ collagen mRNA in situ hybridization histochemistry staining were applied.MAIN OUTCOME MEASURES: Results of HE staining, Masson staining and in situ hybridization histochemistry staining.RESUTLS: A total of 20 animals were included in the result analysis. ①HE staining showed that inflammatory cell infiltration and significant infiltrative zone of leukocytes occurred in hyperplastic scars of right ears in each group. There was inflammatory cell infiltration, but no infiltrative zone of leukocytes in hyperplastic scars of left ears after intervention with antisense TGF β1. ②Masson staining suggested that collagen fibers with deep blue-stain occurred in hyperplastic scars of right ears from the 3rd week after injury, till the 7th week there still was blue-stain collagen fibers,which was bulky (width of about 8-10 μm) and arranged in a great mess.The blue-stain collagenous fibers also appeared in hyperplastic scars of left ears at the 3rd week after injury by the intervention of antisense TGF β1,but till the 6th and 7th weeks the blue-stain became light and thin (width of about 3-5 μm), arranged in order. ③In situ hybridization revealed that expressive rates of TGF β1 mRNA, type Ⅰ collagen mRNA, type Ⅲ collagen mRNA positive cells decreased obviously.CONCLUSION: Antisense TGF β1 can inhibit the proliferation of hyperplastic scars of rabbits' ears and lighten markedly the fibrosis of cicatricial tissue. The local injection with naked DNA is feasible in the treatment of cicatricle.