中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2014年
1期
117-120
,共4页
博来霉素%乳腺退化蛋白-39%转化生长因子-β1%肺纤维化
博來黴素%乳腺退化蛋白-39%轉化生長因子-β1%肺纖維化
박래매소%유선퇴화단백-39%전화생장인자-β1%폐섬유화
Bleomycin%Breast regression protein-39%Transforming growth factor-β1%Lung fibrosis
目的 观察博来霉素诱导小鼠肺组织纤维化过程中乳腺退化蛋白-39(BRP-39)和转化生长因子-β1(TGF-β1)的表达随时间的变化水平,探讨BRP-39和TGF-β1在肺纤维化病理改变中的作用.方法 采用气管滴入5 g/L博来霉素溶液25μl制备肺纤维化模型,在造模后7、14、21 d分别留取肺组织并应用苏木素-伊红(HE)和Masson染色,观察肺组织纤维化改变；酶联免疫吸附试验(ELISA)法检测羟脯氨酸含量变化水平；免疫组织化学法和Westem blot法分别检测TGF-β1和BRP-39表达.结果 博来霉素气管滴人可诱导小鼠肺组织纤维化随时间的增加而持续加重,第21天最为明显,HE和Masson染色均可见肺组织明显纤维化病灶.各时间点的肺组织羟脯氨酸检测含量值均明显高于对照组,且于第14天达峰值(P＜0.05).免疫组织化学检测结果表明,TGF-β1广泛分布于炎性细胞和纤维化肺组织且表达水平随时间进展而增加,在第7天时达到高峰(68.54±2.36,P＜0.05).Western blot结果显示,BRP-39表达含量显著增加(P＜0.05),并在造模后第14天达到峰值(0.790 ±0.042).结论TGF-β1和BRP-39与肺纤维化的发生密切相关,并在肺纤维化进展中起重要作用.
目的 觀察博來黴素誘導小鼠肺組織纖維化過程中乳腺退化蛋白-39(BRP-39)和轉化生長因子-β1(TGF-β1)的錶達隨時間的變化水平,探討BRP-39和TGF-β1在肺纖維化病理改變中的作用.方法 採用氣管滴入5 g/L博來黴素溶液25μl製備肺纖維化模型,在造模後7、14、21 d分彆留取肺組織併應用囌木素-伊紅(HE)和Masson染色,觀察肺組織纖維化改變；酶聯免疫吸附試驗(ELISA)法檢測羥脯氨痠含量變化水平；免疫組織化學法和Westem blot法分彆檢測TGF-β1和BRP-39錶達.結果 博來黴素氣管滴人可誘導小鼠肺組織纖維化隨時間的增加而持續加重,第21天最為明顯,HE和Masson染色均可見肺組織明顯纖維化病竈.各時間點的肺組織羥脯氨痠檢測含量值均明顯高于對照組,且于第14天達峰值(P＜0.05).免疫組織化學檢測結果錶明,TGF-β1廣汎分佈于炎性細胞和纖維化肺組織且錶達水平隨時間進展而增加,在第7天時達到高峰(68.54±2.36,P＜0.05).Western blot結果顯示,BRP-39錶達含量顯著增加(P＜0.05),併在造模後第14天達到峰值(0.790 ±0.042).結論TGF-β1和BRP-39與肺纖維化的髮生密切相關,併在肺纖維化進展中起重要作用.
목적 관찰박래매소유도소서폐조직섬유화과정중유선퇴화단백-39(BRP-39)화전화생장인자-β1(TGF-β1)적표체수시간적변화수평,탐토BRP-39화TGF-β1재폐섬유화병리개변중적작용.방법 채용기관적입5 g/L박래매소용액25μl제비폐섬유화모형,재조모후7、14、21 d분별류취폐조직병응용소목소-이홍(HE)화Masson염색,관찰폐조직섬유화개변；매련면역흡부시험(ELISA)법검측간포안산함량변화수평；면역조직화학법화Westem blot법분별검측TGF-β1화BRP-39표체.결과 박래매소기관적인가유도소서폐조직섬유화수시간적증가이지속가중,제21천최위명현,HE화Masson염색균가견폐조직명현섬유화병조.각시간점적폐조직간포안산검측함량치균명현고우대조조,차우제14천체봉치(P＜0.05).면역조직화학검측결과표명,TGF-β1엄범분포우염성세포화섬유화폐조직차표체수평수시간진전이증가,재제7천시체도고봉(68.54±2.36,P＜0.05).Western blot결과현시,BRP-39표체함량현저증가(P＜0.05),병재조모후제14천체도봉치(0.790 ±0.042).결론TGF-β1화BRP-39여폐섬유화적발생밀절상관,병재폐섬유화진전중기중요작용.
Objective To explore the expression of breast regression protein-39 (BRP-39) and transforming growth factor-β1 (TGF-β1) over time in mice with bleomycin-induced lung fibrosis,and initially reveal the pathological role of BRP-39 and TGF-β1 in pulmonary fibrosis.Methods Pulmonary fibrosis model was prepared by tracheal instillation with 5 g/L solution of 25 μl bleomycin.After 7,14 and 21 d,the lung tissue specimens were stained with hematoxylin and eosin (HE) and Masson staining,and lung tissue fibrosis was observed.Enzyme linked immunosorbent assay (ELISA) was applied to detect the contents of hydroxyproline.Immunohistochemistry and Western blotting were used to detect the expression of TGF-β31 and BRP-39.Results Tracheal instillation of bleomycin induced pulmonary fibrosis which was aggravated over time and most obvious at 21 d.HE and Masson staining displayed the lung tissue fibrosis lesions.Hydroxyproline contents in the fibrotic lung tissue were significantly higher than in the controls at each time point,and reached the peak at day 14 (P ＜ 0.05).Immunohistochemistry revealed that TGF-β1 was widely distributed in inflammatory cells and fibrotic lung tissue and its expression level was increased over time,and reached its peak at day 7 (P ＜ 0.05).Western blotting demonstrated that BRP-39 expression in the fibrotic lung tissue was significantly increased (P ＜ 0.05),and reached its peak at day 14.Conclusion Both TGF-β1 and BRP-39 are closely related to the occurrence of pulmonary fibrosis and may play an important role in pulmonary fibrosis.
