霉酚酸酯对小鼠肺纤维化组织中转化生长因子-β1表达的影响  

Effect of mycophenolate mofetil on transforming growth factor-β1 gene expression in lung tissue of mice with pulmonary fibrosis

作　　者：曹姝[1,2] 李少林[2] 吴府容[1] 康强强[2] 李芳[2] 王颖[1] 

机构地区：[1]重庆市肿瘤研究所,重庆400030 [2]重庆医科大学放射医学教研室,重庆400016

出　　处：《中国生物制品学杂志》2013年第7期974-978,共5页Chinese Journal of Biologicals

基　　金：重庆市科委资助项目(CSTC;2009BB5059)

摘　　要：目的探讨霉酚酸酯(mycophenolate mofetil,MMF)对博莱霉素(bleomycin,BLM)诱导的小鼠肺纤维化组织中转化生长因子-β1(transforming growth factor-β1,TGF-β1)表达的影响。方法将C57BL/6小鼠随机分为6组:正常对照组、MMF对照组、BLM模型组、MMF低(20 mg/kg)、中(60 mg/kg)、高(100 mg/kg)剂量干预组,每组6只。BLM模型组和MMF 3个干预组经气管注入BLM(6 mg/kg),正常对照组和MMF对照组注入等量无菌生理盐水;次日按MMF对照组和MMF干预低、中、高剂量组小鼠体重计算MMF给药量,灌胃小鼠,每日1次,连续14 d,正常对照组和BLM模型组用等体积的双蒸水灌胃。灌胃第16天采集小鼠肺脏标本,经HE、Masson染色,从组织形态学上观察肺组织纤维化情况并进行Aschcroft评分;RT-PCR法检测小鼠肺组织中TGF-β1基因mRNA的转录水平;Western blot法检测小鼠肺组织中TGF-β1蛋白的表达水平。结果 BLM诱导的小鼠肺纤维化改变显著,Ashcroft评分较正常对照组显著增高(P<0.01),表明小鼠肺纤维化模型构建成功;MMF高剂量干预组肺组织损伤减轻,炎细胞浸润及胶原沉积减少,Ashcroft评分显著降低(P=0.000)。MMF高剂量干预组与BLM模型组比较,TGF-β1基因mRNA的转录水平及蛋白的表达水平均明显减低(P<0.05);而MMF低、中剂量干预组与BLM模型组之间、MMF对照组与正常对照组之间,差异均无统计学意义(P>0.05)。结论高剂量的MMF(100 mg/kg)可抑制BLM诱导的肺纤维化小鼠肺组织中TGF-β1基因mRNA的转录水平及蛋白表达水平,有望成为治疗肺纤维化的理想药物。Objective To investigate the effect of mycophenolate mofetil（MMF） on transforming growth factor-β1（TGF-β1） gene expression in bleomycin（BLM）-induced mouse model of pulmonary fibrosis.Methods C57BL / 6 mice were randomly divided into six groups,six for each,i.e.normal control,MMF control,BLM model as well as low（20 mg / kg）,moderate（60 mg / kg） and high（100 mg / kg） dose MMF treatment groups.The mice in BLM model and three MMF treatment groups were intratracheally instilled with BLM（6 mg / kg）,while those in normal control and MMF control groups with sterile physiological saline at an equal volume.One day after instillation,the mice in MMF control and MMF treatment groups were treated with MMF by lavage at the calculated dosages based the bodyweight,once a day for 14 d,while those in normal control and BLM model groups with double distilled water at an equal volume.Lung specimens of mice were collected on day 16 after lavage,observed for pulmonary fibrosis by HE and Masson staining and subjected to Aschcroft scoring.The TGF-β1 mRNA transcription level in lung tissue was determined by RT-PCR,while the protein expression level by Western blot.Results BLM-induced pulmonary fibrosis in mouse model was improved significantly,of which the Ashcroft score was significantly higher than that in normal control group（P 0.01）,indicating that the mouse model of pulmonary fibrosis was established successfully.The lung tissue lesion of mice in high dose MMF treatment group was relieved,while the inflammatory cell infiltration and collagen deposition decreased,and the Ashcroft score decreased significantly（P = 0.000）.Both the mRNA transcription and protein expression levels of TGF-β1 was significantly lower in high dose MMF treatment group than in BLM model group（P 0.05）,while showed no significant difference in low and moderate dose MMF treatment and BLM model groups,or in MMF control and normal control groups（P 0.05）.Conclusion High dose（100 mg / kg） MMF significantly inhibited

关 键 词：霉酚酸酯 博来霉素 肺纤维化 转化生长因子-Β1 

分 类 号：R979.5[医药卫生—药品] R563.9[医药卫生—药学]