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目的观察自噬相关蛋白LC-3和Beclin-1在大鼠硅沉着病模型中的表达及意义,并探讨其机制;从细胞自噬角度研究硅沉着病形成的分子机制。方法将90只健康SD雄性大鼠随机分为对照组、模型组和自噬抑制剂3-甲基腺嘌呤(3-MA)组,每组30只。采用非暴露式气管插管一次性灌注二氧化硅粉尘混悬液法建立大鼠硅沉着病模型,并给予3-MA干预,各组分别于造模后第1、3、7、14及28天分批处死6只大鼠。留取肺组织,HE和Masson染色观察肺组织形态变化;免疫组织化学染色检测LC-3、Beclin-1的表达及分布;免疫印迹法检测LC-3、Beclin-1的蛋白表达量。结果与对照组相比,模型组有显著的肺泡炎改变、明显的矽结节形成及大量胶原沉积。LC-3、Beclin-1在各时间点的表达均上调(P<0.05),并随时间呈现动态变化趋势,SiO2注入后1 d表达有升高趋势,14d达高峰(P<0.05),28 d回落,但仍高于基础表达。与模型组相比,3-MA组肺泡炎减轻,矽结节减小,胶原沉积减少,LC-3、Beclin-1在各时间点的表达均下调(P<0.05),各时间点的变化趋势无显著性改变。结论细胞自噬参与大鼠硅沉着病的病理进程,并在硅沉着病的发生与发展过程中起重要作用。
Objective To investigate the expression and significance of autophagy-related proteins LC-3 and Beclin-1 in rat model of silicosis and to explore their mechanism. To study the molecular mechanism of the formation of silicosis from the angle of autophagy. Methods Ninety healthy SD male rats were randomly divided into control group, model group and autophagy inhibitor 3-methyladenine (3-MA) group, 30 rats in each group. Non-exposed endotracheal intubation with silicon dioxide dust suspension perfusion method to establish rat model of silicosis, and given 3-MA intervention, each group were in the modeling of the first 1,3,7,14 and 28 Toxicity dead 6 rats. The lung tissues were collected and the morphological changes of lung were observed by HE and Masson staining. The expression and distribution of LC-3 and Beclin-1 were detected by immunohistochemical staining. The protein expression of LC-3 and Beclin-1 were detected by Western blotting. Results Compared with the control group, the model group had significant changes of alveolitis, obvious formation of silicon nodules and massive collagen deposition. The expression of Beclin-1 and Beclin-1 at each time point were up-regulated (P <0.05), and showed a trend of dynamic change with time. The expression of Beclin-1 and Beclin- d down, but still higher than the basic expression. The expression of LC-3 and Beclin-1 were down-regulated at all time points (P <0.05) in 3-MA group compared with the model group, the alveolitis decreased, the number of silicon nodules and collagen deposition decreased in 3-MA group No significant change in trend. Conclusion Cell autophagy is involved in the pathogenesis of silicosis in rats and plays an important role in the occurrence and development of silicosis.