他克莫司预防兔耳瘢痕增生的实验研究

Preventive effects of Tacrolimus on the rabbit ear hypertrophic scar model

目的:研究他克莫司对兔耳瘢痕增生的影响。方法:选10只新西兰白兔雌雄各半,按本研究组建立的改良方法制作兔耳瘢痕模型,左耳为空白对照组涂凡士林软膏,右耳为实验组涂他克莫司软膏。伤后14天、21天、28天、35天及49天采集标本,行苏木精-伊红(HE)及Masson三色法染色,统计成纤维细胞密度,实时定量PCR(Real-time PCR)检测细胞外基质成分Ⅰ型胶原(collagen Ⅰ;ColI)和纤维连接蛋白(fibronectin;Fn),CD4+辅助性T细胞-2(Th2)产生的重要纤维化基因IL-4及参与T细胞早期活化的重要基因巨噬细胞集落刺激因子(macrophage colony stimulating factor;M-CSF)和肿瘤坏死因子-α(tumor necrosis factorα;TNF-α)的表达。结果:HE及Masson三色法染色可见实验组较对照组胶原沉积明显减少,PCR结果可见ColI、Fn、IL-4、M-CSF和TNF-α在实验组中的表达较对照组在各时间点均减少。结论:应用他克莫司可通过下调IL-4、M-CSF和TNF-α的表达来抑制兔耳瘢痕增生。

Objective To evaluate the preventive effect of tacrolimus on rabbit ear hypertrophic scar formation.Methods Ten New Zealand White rabbits were employed in this study.Ventral ear hypertrophic scar model was created according to our previous report.Experimental group contains all the right ear wounds and treated with tacrolimus ointment.The control group contains all the left ear wounds and treated with vaseline ointment.Both the groups were harvested on days 14,21,28,35 and 49 post wounding.Hematoxylin-eosin staining and Masson trichrome staining were performed to reveal differences in scar morphology.Real-time PCR was adopted to evaluate the expression of ColI,Fn,IL-4,M-CSF and TNF-α.Results The morphology had a large difference in the two groups.Compared with the control group,the fibroblast number in the treated group was significantly reduced（P0.01）.The expression of ColI,Fn,IL-4,M-CSF and TNF-α were distinctly down-regulated in the treatment group.Conclusion Dealing with the post-wounding surface with tacrolimus may effectively suppress hypertrophic scar formation through down-regulating the expression of IL-4.、M-CSF and TNF-α.