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目的:探讨干扰小RNA(siRNA)下调瘦素(leptin)的表达对体外培养的人病理性瘢痕成纤维细胞增殖能力及转化生长因子-βn 1(TGF-βn 1)、Ⅰ型胶原蛋白表达水平的影响。n 方法:收集川北医学院附属医院烧伤整形科手术切除的病理性瘢痕组织(增生性瘢痕、瘢痕疙瘩)。经原代细胞培养及细胞传代，取第3代细胞用于实验研究。实验分为2组：转染leptin-siRNA为实验组；转染空载体为阴性对照组。细胞转染48 h后进行后续实验。以CCK-8法测定细胞增殖能力。以实时定量聚合酶链反应(PCR)检测leptin、TGF-βn 1、Ⅰ型胶原蛋白的mRNA表达水平。以免疫荧光法和Western blotting测定leptin、TGF-βn 1、Ⅰ型胶原蛋白的蛋白表达量。组间比较采用n t检验，n P0.05)。增生性瘢痕及瘢痕疙瘩组织中，leptin、TGF-βn 1以及Ⅰ型胶原蛋白的mRNA相对表达量比较，实验组比阴性对照组均明显减少(n P<0.05)。免疫荧光显示，瘢痕疙瘩中leptin、TGF-βn 1、Ⅰ型胶原蛋白的表达高于增生性瘢痕，且应用siRNA下调leptin后，增生性瘢痕及瘢痕疙瘩成纤维细胞中leptin、TGF-βn 1、Ⅰ型胶原蛋白表达明显减少。Western blotting检测结果示增生性瘢痕和瘢痕疙瘩成纤维细胞中leptin、TGF-βn 1和Ⅰ型胶原蛋白在siRNA干扰下显著降低，其转染siRNA组的leptin、TGF-βn 1、Ⅰ型胶原蛋白表达水平均小于阴性对照组(n P均值<0.01)。n 结论:RNA干扰下调leptin基因的表达可抑制TGF-βn 1和Ⅰ型胶原蛋白的表达，有希望用于病理性瘢痕的治疗。n “,”Objective:To investigate the effects of RNA interference on the expression level of leptin and its effect on proliferation, TGF-βn 1 and collagen type Ⅰ production in human pathological scar fibroblasts n in vitro.n Methods:Pathological scar tissues (hypertrophic scar, keloid) were collected from the Department of Burn Plastic Surgery in Affiliated Hospital of North Sichuan Medical College. After primary cell culture and cell passage, passage 3 cells were selected for experimental study. The cells were divided into two groups: the experimental group which was transfected with leptin siRNA, and the negative control group which was transfected with empty vector. Examinations were carried out 48 hours after transfection. Cell proliferation was determined by CCK-8. The transcription levels of leptin, TGF-βn 1 and type Ⅰ collagen genes were detected by polymerase chain reaction (PCR). The protein expression levels of leptin, TGF-βn 1 and type Ⅰ collagen were determined by immunofluorescence and Western blotting. Student′s n t-test was used for comparison between groups, and n P0.05). The relative mRNA expression levels of leptin, TGF-βn 1 and type Ⅰ collagen in hypertrophic scars and keloids were significantly decreased in the siRNA transfection groups compared with the negative control groups (n P<0.05). Immunofluorescence results showed that the expression levels of leptin, TGF-βn 1 and type I collagen in keloids were higher than those in hypertrophic scars, and that siRNA induced leptin downregulation significantly reduced the levels of leptin, TGF-βn 1 and type I collagen in both hypertrophic scars and keloids. Western blotting showed that the protein levels of leptin, TGF-βn 1 and type Ⅰ collagen were significantly decreased in hypertrophic scar and keloid fibroblasts after leptin siRNA interference (n P<0.05).n Conclusions:Downregulation of leptin gene expression by RNA interference inhibited TGF-βn 1 and typeⅠ collagen expression, which could be used in treating pathological scar.n