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目的:探讨博来霉素诱导小鼠肺间质纤维化过程中乳腺退化蛋白39(BRP-39)的表达水平变化。方法:取7-8周龄C57BL/6小鼠20只,随机分为博来霉素模型组和正常对照组,每组各10只;博来霉素组采用气管滴入5g/L博来霉素溶液25μl,而对照组则予以等量生理盐水。处理后14d,分别取两组小鼠肺组织应用HE和Masson染色,观察肺组织改变情况;ELISA法检测羟脯氨酸含量;免疫组织化学法和Western blot法分别检测BRP-9蛋白表达水平。结果:博来霉素处理后,镜下HE染色可见肺泡间隔轻度增宽、病灶区域巨噬细胞等炎性细胞浸润的改变,且肺组织损伤分布较广,纤维沉积明显;且与正常对照组比较,Masson蓝染胶原也明显增多;此外,博来霉素组小鼠肺组织中羟脯氨酸含量明显高于正常对照组(P<0.05);免疫组织化学检测结果表明,博来霉素组BRP-39广泛分布于炎症细胞和纤维化肺组织中且阳性表达水平显著高于正常对照组(P<0.05);Western blot结果亦显示,肺间质纤维化可引起BRP-39表达含量显著增加,蛋白表达量可达正常对照组的2.35倍(P<0.05)。结论:BRP-39在肺纤维化模型中升高,提示其可能在肺间质纤维化过程中发挥重要作用。
Objective: To investigate the expression of breast degeneration protein 39 (BRP-39) in bleomycin induced pulmonary interstitial fibrosis in mice. Methods: Totally 20 C57BL / 6 mice aged 7-8 weeks were randomly divided into bleomycin model group and normal control group, with 10 rats in each group. Bleomycin group was treated with 5 g / 25 microliters of mycophenolate solution, while the control group was given the same amount of normal saline. At 14 days after treatment, the lungs of the two groups were harvested and stained with HE and Masson to observe the changes of lung tissue. The contents of hydroxyproline were detected by ELISA. The expressions of BRP-9 protein were detected by immunohistochemistry and Western blot respectively. Results: After bleomycin treatment, the alveolar septum slightly widened, inflammatory cell infiltration of macrophages in the lesion area was observed by microscopic HE staining, and the damage of lung tissue was extensively distributed and the fiber deposition was obvious. Compared with the normal control In addition, the content of hydroxyproline in lung tissue of bleomycin group was significantly higher than that of normal control group (P <0.05). The results of immunohistochemistry showed that the percentage of bleomycin BRP-39 was widely distributed in inflammatory cells and fibrosis lung tissue and the positive expression level was significantly higher than the normal control group (P <0.05); Western blot results also showed that pulmonary fibrosis can cause BRP-39 expression (P <0.05), which was significantly higher than that of control group (P <0.05). Conclusion: BRP-39 is elevated in pulmonary fibrosis model, suggesting that BRP-39 may play an important role in the process of pulmonary fibrosis.