CAJ | 학술논문

背景:瘢痕的形成涉及多种因素,其中之一是转化生长因子β1(TGFβ1)的高表达,为达到抑制TGFβ1过度表达的目的,尝试用反义TGFβ1抑制瘢痕形成的方法,将基因治疗技术运用于临床医学.目的:探讨反义TGFβ1对Ⅲ度烫伤愈合中瘢痕生成的抑制作用.设计:随机对照、重复测量设计.地点和对象:在上海第二医科大学生物化学教研室完成.清洁级雄性SD大鼠105只,体质量(250±20)g,购于中科院上海实验动物中心.干预:SD大鼠Ⅲ度烫伤后,分3组进行处理:①反义TCFβ1脱氧寡核苷酸.②反义TGFβ1重组质粒.③空白对照.分别于烫伤后1,3,5,14 dRT-PCR、免疫组化检测TGFβ1mRNA和蛋白质的表达.烫伤后5,14,21,28,60 d原位杂交检测Ⅰ型胶原mRNA的表达变化,病理检测炎性细胞浸润反应和胶原分布状况.主要观察指标:①烫伤后1,3,5,14 d RT-PCR,TGFβ1mRNA和蛋白质的表达.②烫伤后5,14,21,28,60 d Ⅰ型胶原mRNA的表达变化.③病理检测炎性细胞浸润反应和胶原分布状况.结果:反义ODN组和反义重组质粒组在烫伤后第1天TGFβ1mRNA表达水平与对照组相比无降低,14 d分别为0.176±0.014和0.213±0.071,表达均减少.反义作用组TGFβ1蛋白质的表达在烫伤后3周内均为低表达.原位杂交显示对照组在14 dⅠ型胶原mRNA开始表达,28 d达到高峰,随后减少,反义作用组无此现象,一直保持低表达.病理染色显示反义作用组炎性细胞浸润少,炎性反应低,胶原合成减少.结论:反义TGFβ1可抑制Ⅲ度烫伤愈合中瘢痕的形成.
배경:반흔적형성섭급다충인소,기중지일시전화생장인자β1(TGFβ1)적고표체,위체도억제TGFβ1과도표체적목적,상시용반의TGFβ1억제반흔형성적방법,장기인치료기술운용우림상의학.목적:탐토반의TGFβ1대Ⅲ도탕상유합중반흔생성적억제작용.설계:수궤대조、중복측량설계.지점화대상:재상해제이의과대학생물화학교연실완성.청길급웅성SD대서105지,체질량(250±20)g,구우중과원상해실험동물중심.간예:SD대서Ⅲ도탕상후,분3조진행처리:①반의TCFβ1탈양과핵감산.②반의TGFβ1중조질립.③공백대조.분별우탕상후1,3,5,14 dRT-PCR、면역조화검측TGFβ1mRNA화단백질적표체.탕상후5,14,21,28,60 d원위잡교검측Ⅰ형효원mRNA적표체변화,병리검측염성세포침윤반응화효원분포상황.주요관찰지표:①탕상후1,3,5,14 d RT-PCR,TGFβ1mRNA화단백질적표체.②탕상후5,14,21,28,60 d Ⅰ형효원mRNA적표체변화.③병리검측염성세포침윤반응화효원분포상황.결과:반의ODN조화반의중조질립조재탕상후제1천TGFβ1mRNA표체수평여대조조상비무강저,14 d분별위0.176±0.014화0.213±0.071,표체균감소.반의작용조TGFβ1단백질적표체재탕상후3주내균위저표체.원위잡교현시대조조재14 dⅠ형효원mRNA개시표체,28 d체도고봉,수후감소,반의작용조무차현상,일직보지저표체.병리염색현시반의작용조염성세포침윤소,염성반응저,효원합성감소.결론:반의TGFβ1가억제Ⅲ도탕상유합중반흔적형성.
BACKGROUND: Formation of scar is related to various factors, of which is the high expression of transforming growth factor β1 (TGFβ1). In order to inhibit the overexpression of TGFβ1, anti-TGFβ1 is used to inhibit the scar formation for the application of gene therapeutic techniques in clinical medicine.OBJECTIVE: To explore the inhibitory effects of anti-TGFβ1 on scar formation during healing of third-degree burns.DESIGN: A randomized controlled repeated measures design.SETTING and PARTICIPANTS: The study was completed in the Department of Biochemistry, Shanghai Second Medical University. The 105 male SD rats(clean grade) weighing 250 ±20 g were purchased from Shanghai Center for Experimental Animals.INTERVENTIONS: A total of 105 SD rats were divided into three groups after being scalded to third-degree bums: anti-TGFβ1 oligodeoxyribonucleotides(anti-TGFβ1 ODN) group, anti-TGFβ1 recombinant plasmid group and blank control group. On the 1st, 3rd, 5th and 14th day after burn, RT-PCR and immunohistochemical methods were used to test the expression of TGFβ1mRNA and protein. On the 5th, 14th, 21st, 28th and 60th day after burn, in situ hybridization was used to detect the changes of type Ⅰ collagen mRNA expression, inflammatory cell infiltration and distribution of collagen were determined by pathological test.The changes of type Ⅰ collagen mRNA expression on the 5th, 14th, 21st , 28th collagen were detected by pathological test.RESULTS: As compared with control group, there was no obvious change in expression level of TGFβ1 mRNA in anti-TGFβ1 ODN group and anti-TGFβ1recombinant plasmid group on the 1st day after scald. On the 14th day, the expression decreased, and was 0. 176 ±0. 014 and 0. 213 ±0. 071 in anti-TGFβ1 ODN group and anti-TGFβ1 recombinant plasmid group, respectively. The expression pf TGF β1 protein was reduced in the experimental groups during the first three weeks after scald. The in situ hybridization test showed that there was type I collagen mRNA expressed in the control group since the 14th day and reached peak on the 28th day and then decreased. No same phenomena were found in the experimental groups, but kept low expression. Pathological staining showed that there were a few infiltrated inflammatory cells, low inflammatory reaction and reduced collagen synthesis in the experimental groups.CONCLUSION: Anti-TGFβ1 can inhibit the scar formation in the healing of third-degree bums.