摘要
目的观察博莱霉素致小鼠肺间质纤维化中解整合素-金属蛋白酶17(a disintegrin and metalloproteinase-17,ADAM17)表达水平变化与转化生长因子-β1(transforming growth factor-β1,TGF-β1)之间的相关性,以阐明肺纤维化的发病机制。方法 60只小白鼠随机分为6组,每组10只。正常对照组经气管内一次性注入0.9%氯化钠注射液(5 mg/kg)。模型组5组分别为注射博莱霉素3 d、7 d、14 d、21 d、28 d组,经气管内一次性注入博莱霉素(5 mg/kg),分别在规定的时间点取肺组织,通过Masson胶原染色评价肺纤维化的程度,用免疫组织化学染色和蛋白质印迹(Western blotting)方法检测ADAM17和TGF-β1在肺组织的蛋白水平表达,RT-PCR检测ADAM17和TGF-β1 mRNA的表达。结果博莱霉素诱导小鼠肺纤维化模型成功,模型组病理呈不同程度的胶原沉积和成纤维细胞增生。小鼠纤维化程度自7 d组开始均高于正常对照组(P<0.01)。免疫组化结果示,TGF-β1、ADAM17在对照组呈弱表达,模型组表达量高于对照组(P<0.01),且14 d时表达量最高。Western blotting和RT-PCR结果与免疫组化结果一致。相关性分析结果示,TGF-β1、ADAM17在免疫组化、Western blotting和RT-PCR中的相关系数r分别为0.777、0.808、0.906(P<0.01),表明二者呈正相关。结论 ADAM17在博莱霉素致小鼠肺间质纤维化过程中表达水平上调,ADAM17可能通过上调TGF-β1发挥致纤维化的作用。
Objective To study the expression level of a disintegrin and metalloproteinase-17 （ADAM 17） on bleomycin-induced pulmonary fibrosis in rats, investigate the correlation between ADAM17 and transforming growth factor-β11 （TGF- β1）, and illustrate the pathogenesis of pulmonary fibrosis. Methods Sixty rats were randomly divided into six groups, 10 in each group. In the control group, the rats were treated with single intratracheal instillation and injected with 0.9% sterile saline solution （5mg/kg）. The model group were divided into 3rd day group, 7th day group, 14th day group, 21th day group and 28th day group. They were all injected with bleomycin （5 mg/kg） and their lung tissues were taken at specified time point respectively. Histological changes of the lung tissues were evaluated by Massson＇s stain. The expression of ADAM17 and TGF- β1 proteins was assessed by immunohistochemistry stain and Western blotting, and the expression of their mRNA was detected by reverse transcription-polymerase chain reaction （RT-PCR）. Results Bleomycin-induced pulmonary fibrosis in rats was successfully established, the pathological changes of model groups showed varying degrees of collagen deposition and myofibroblast proliferation. The level of fibrosis in model groups （except the 3rd day group） were higher than those in control group （P 〈 0.01）. According to immunohistochemistry data, the expressions of TGF- β1 and ADAM17 were higher than those in control group （P 〈 0.01）, and both of them were the highest in the 14th day group. The results of Western blotting and RT-PCR were mainly consistent with the immunohistochemical results. Correlation analysis pointed out that TGF- β1 was positively correlated with ADAMI 7, the correlation coefficients in immunohistochemistry, Western blotting and RT-PCR were 0.777, 0.808, 0.906, respectively （P 〈 0.01）. Conclusion The expression level of ADAM17 increases in bleomycin- induced pulmonary fibrosis in rats, ADAMI7 may play the role of inducing pulmonary fibrosis by increasing the expression of TGF- β1.
出处
《解放军医学院学报》
CAS
2014年第9期947-951,960,共6页
Academic Journal of Chinese PLA Medical School
基金
辽宁省自然科学基金项目(201102133)~~
