丝裂霉素C预防家兔喉气管狭窄的实验研究

Mitomycin C prevents laryngotracheal stenosis in rabbits

目的探讨丝裂霉素C在治疗兔喉气管狭窄中的作用及其机制。方法将30只家兔按照简单随机化分组分为对照组(A组)、模型组(B组)、0.5 mg/m L低浓度用药组(C1组)、2 mg/m L中浓度用药组(C2组)、5 mg/m L高浓度用药组(C3组)。制备喉气管狭窄模型,用药后20d比较各组喉气管狭窄度。取模型组狭窄组织做成纤维细胞培养,分别置于0、2.5、12.5、50、100、200 mg/L的丝裂霉素C中培养后用流式细胞仪检测细胞凋亡差异,Western blot、荧光定量PCR检测细胞b FGF、TGF-β1的蛋白和mRNA转录水平表达变化。结果 B、C组相比A组均出现明显喉气管狭窄(P<0.05),C2、C3 2组狭窄度明显低于B组(P<0.05),C1组与B组无明显差异(P>0.05);丝裂霉素C能诱导成纤维细胞凋亡,下调TGF-β1、b FGF的蛋白和mRNA表达量。结论丝裂霉素C可明显诱导成纤维细胞凋亡,对预防和治疗瘢痕性喉气管狭窄具有重要作用。

Objective To explore the effect of mitomycin C on laryngotracheal stenosis in rabbits.Methods Rabbits were randomly divided into 5 groups,that is,a control group（group A）,a model group（group B）,a 0.5 mg /m L low-dose group（group C1）,a 2 mg /m L middle-dose group（group C2） and a5 mg / m L high-dose group（group C3）.The model of rabbit laryngotracheal stenosis was established by scraping the laryngotracheal mucosa.All rabbits in group C1,group C2 and group C3 received topical application of mitomycin C to the wound surface of larygotracheal mucosa.The laryngotracheal stenosis degree was observed on the 20 th day after operation.The fibroblasts taken from the stenosis tissue of the model group were cultured in vitro with mitomycin C at 6 different concentrations（0,2.5,12.5,50,100 mg / m L,and 200 mg / m L）.Meanwhile,the fibroblast apoptosis difference was detected by flow cytometry.In addition,the mRNA and protein expression of basic fibroblast growth factor（b FGF） and transforming growth factor β1（TGF-β1） in the6 groups was detected by FQ-PCR and Western blotting,respectively.Results Compared with group A,both group B and group C showed obvious laryngotracheal stenosis（P 〈 0.05）.The stenosis degree of group C2 and group C3 was significantly lower than that of group B（P 〈 0.05）,but there was no significant difference between group B and group C1（P 〉 0.05）.Mitomycin C could significantly inhibit the proliferation of the fibroblasts and reduce the mRNA and protein expression of TGF-β1 and b FGF.Conclusion Mitomycin C can significantly induce the apoptosis of fibroblasts,and may play an important role in preventing and treating laryngotracheal stenosis.