转化生长因子-β1与系统性硬化病-肺纤维化相关性的实验研究

Correlation between Transforming Growth Factor-β1 and Systemic Sclerosis-Pulmonary Fibrosis

目的探讨转化生长因子-β1(TGF-β1)在系统性硬化病-肺纤维化(SSc-PF)模型小鼠中的表达及相关性。方法雄性小鼠30只,随机分为磷酸盐缓冲液皮下注射组(A组)10只和博来霉素皮下注射组(模型组)20只。根据肺组织Ashcroft半定量评分将模型组小鼠分为B组11只(Ashcroft半定量评分≥3分)和C组9只(Ashcroft半定量评分<3分)。A组皮下注射磷酸盐缓冲液,B组、C组皮下注射博来霉素建立小鼠SSc-PF模型。应用马松三色染色和苏木素-伊红染色观察小鼠肺组织病理学和形态学改变;反转录-聚合酶链反应、酶联免疫吸附测定法检测3组小鼠外周血TGF-β1、肺组织TGF-β1 mRNA含量;采用样本碱水解法测定肺组织羟脯氨酸含量,并进行肺组织Ashcroft半定量评分,分析这些指标间的相关性。结果 B组肺组织Ashcroft半定量评分、羟脯氨酸含量及外周血TGF-β1、肺组织TGF-β1 mRNA含量明显高于A组、C组(P<0.05)。但A组、C组肺组织Ashcroft半定量评分、羟脯氨酸含量及肺组织TGF-β1 mRNA、外周血TGF-β1含量比较,差异无统计学意义(P>0.05)。肺组织TGF-β1 mRNA、外周血TGF-β1含量与肺组织Ashcroft半定量评分、羟脯氨酸含量均呈正相关关系(P<0.05)。结论 TGF-β1在SSc-PF小鼠模型中高表达,并随SSc-PF病变程度增加而增加。

Objective To investigate transforming growth factor-β1（ TGF-β1） expression in the model mice with systemic sclerosis-pulmonary fibrosis（ SSc-PF） and its correlation with SSc-PF. Methods Thirty male mice were randomly divided into Group A（ 10 mice） and model group（ 20 mice）. The mice in Group A were given subcutaneous injection of phosphate buffer while in the model group were given subcutaneous injection of bleomycin. The mice in the model group were divided into Group B（ Ashcroft semi-quantitative score≥3,11 mice） and Group C（ Ashcroft semi-quantitative score〈 3,9 mice）. The mice in Group B and Group C were injected with bleomycin to establish SSC-PF mouse model. Masson staining and HE staining were used to observe the pathological and morphological changes in lung tissues of the mice.Reverse transcription-polymerase chain reaction（ RT-PCR） and enzyme-linked immunosorbent assay（ ELISA） were used to test the contents of TGF-β1,TGF-β1 mRNA in the peripheral blood of lung tissues of the mice in three groups. The alkali hydrolysis was used to detect the content of hydroxyproline in lung tissues,and Ashcroft semi-quantitative scores of lung tissues was calculated. An analysis on the correlation between the indicators was conducted. Results The Ashcroft semi-quantitative score of lung tissues,the contents of hydroxyproline,TGF-β1 in peripheral blood and TGF-β1 mRNA in lung tissues in Group B were significantly higher than those in Group A and Group C（ P〈0. 05）. But the Ashcroft semi-quantitative score of lung tissues,the contents of hydroxyproline,TGF-β1 mRNA in lung tissues and TGF-β1 in peripheral blood showed nosignificant difference between Group A and Group B（ P〉0. 05）. The contents of TGF-β1 mRNA in lung tissues and TGF-β1 in peripheral blood positively correlated with the Ashcroft semi-quantitative score of lung tissues as well as the content of hydroxyproline（ P〈0. 05）. Conclusion TGF-β1 is highly expressed in the model mice with SSc-PF,and increases with the increase of SSc-PF lesion severity.