吡非尼酮抑制兔耳增生性瘢痕作用的研究

Study on the inhibitory effect of pirfenidone on hypertrophic scar in a rabbit ear model

目的探索局部注射吡非尼酮对兔耳增生性瘢痕的抑制作用。方法选取健康新西兰大耳白兔36只,在兔耳双侧分别构建增生性瘢痕模型后,将36只兔子随机平均分为溶剂对照组(A组)、吡非尼酮实验组(B组)和曲安奈德阳性对照组(C组)。待创面完成上皮化后开始给药。B组注射含150μg吡非尼酮的DMSO溶液,共30μl;A组注射等体积的DMSO;C组注射曲安奈德30μl,1次/d,连续注射3 d后改为1次/周,共2次。给药后第45天取材固定,常规HE染色及组织病理学分析,并计算瘢痕增生指数。通过Masson染色分析胶原纤维排列,PCR检测瘢痕组织中TGF-β1、TGF-β2和Col-Ⅰ等基因表达情况。结果 B组和C组与A组相比,吡非尼酮和曲安奈德均可显著降低兔耳瘢痕增生指数,减少瘢痕的高度,其颜色更加接近正常皮肤,胶原组织排列也更为整齐有序;瘢痕组织中TGF-β1、TGF-β2和Col-Ⅰ等的m RNA表达也均明显下降。结论吡非尼酮对兔耳增生性瘢痕的形成具有明显的抑制作用,其初步作用机制可能与抑制瘢痕组织中的TGF-β1和TGF-β2的表达有关。吡非尼酮可能通过下调TGF-β1与TGF-β2等基因的表达抑制了兔耳增生性瘢痕的形成。

Objective To explore the inhibitory effect of local injection of pirfenidone on hypertrophic scar in a rabbit ear model.Methods Thirty-six healthy New Zealand white rabbits were selected.After establishment of hypertrophic scar model in rabbit bilateral ears,36 rabbits were randomly divided into solvent control group(group A),pirfenidone group(group B)and triamcinolone acetonide positive control group(group C).After epithelization of wounds finished,the administration was performed.The DMSO solution containing pirfenidone of 150μg(a total of 30μl)was injected to rabbits in group B.The DMSO of 30μl was injected to rabbits in group A.In group C,triamcinolone acetonide of 30μl was injected to the rabbits once a day.After continuous in jection for 3 days,the administration changed to once a week,a total of twice.At 45 days after administration,scars were collected and fixed.The conventional HE staining and histopathological analysis was performed and the scar elevation index(SEI)was calculated.The arrangement of collagen fibers was observed by Masson staining.The expression of TGF-β1,TGF-β2 and Col-Ⅰin scar tissue was detected by RT-PCR.Results Compared with group A,both pirfenidone and triamcinolone acetonide can significantly reduce the SEI of rabbits and the height of scar.The color was closer to normal skin and the arrangement of collagen tissue was in order.The m RNA expression of TGF-β1,TGF-β2 and ColⅠin the scar tissue was also significantly reduced.Conclusion Pirfenidone has obvious inhibitory effect on the formation of hypertrophic scar in rabbit.The preliminary mechanism might be related to the inhibition of TGF-β1 and TGF-β2 expression in scar.The formation of hypertrophic scar in rabbit ear could be inhibited by pirfenidone through down-regulating the expression of TGF-β1 and TGF-β2.