微管相关蛋白1轻链3及Beclin-1在硅沉着病大鼠肺泡巨噬细胞中的表达

发布时间：2018-03-29 09:18

本文选题：硅沉着病　切入点：肺泡巨噬细胞　出处：《解剖学报》2014年05期

【摘要】：目的观察自噬相关蛋白——微管相关蛋白1轻链3(LC-3)、Beclin-1在硅沉着病大鼠肺泡巨噬细胞中的表达,从细胞自噬角度探讨硅沉着病形成的分子机制。方法 50只成年雄性SD大鼠随机分为2组(n=25):对照组和硅沉着病模型组,每组再分5个时相组。采用非暴露气管灌注二氧化硅(SiO2)粉尘混悬液法(50g/L)建立大鼠硅沉着病模型。分别于造模后1、3、7、14及28 d分批处死5只大鼠,进行原位支气管肺泡灌洗,获取肺泡巨噬细胞,进行培养纯化和富集后用于后续研究。HE染色及透射电子显微镜观察肺泡巨噬细胞形态学改变;免疫细胞化学法检测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肺泡巨噬细胞体积较大,胞质丰富,部分细胞内可见硅沉着吞噬颗粒,电镜下可见自噬体形成;模型组LC-3、Beclin-1在各时间点的表达较对照组均增多(P0.05),1 d即开始增多,随着时间的延长表达逐渐增多,至14 d时达高峰(P0.05),28d时回落,但仍高于对照组的表达。结论在硅沉着病大鼠肺泡巨噬细胞中有自噬的激活,肺泡巨噬细胞自噬参与了大鼠硅沉着病的病理进程。
[Abstract]:Objective to observe the expression of microtubule-associated protein-1 (LC-3LC-3) Beclin-1 in alveolar macrophages of silicosis rats. Methods 50 adult male SD rats were randomly divided into two groups: control group and silicosis model group. Each group was further divided into 5 time phase groups. The silicosis model in rats was established by 50 g / L dust suspension method of non-exposed tracheal instillation of silica, and 5 rats were killed in batches on the 1st day, 714 d and 28 d after the establishment of the model, and then bronchoalveolar lavage was performed in situ. The alveolar macrophages were purified and enriched for further study. He staining and transmission electron microscopy were used to observe the morphological changes of alveolar macrophages and the expression and distribution of LC-3 Beclin-1 were detected by immunocytochemistry. Results compared with the control group, the alveolar macrophages in the model group were larger in volume and abundant in cytoplasm, and the phagocytic particles could be seen in some cells, and autophagy formation was observed under electron microscope. The expression of LC-3 and Beclin-1 in the model group began to increase at 1 day as compared with that in the control group, and gradually increased with the prolongation of the time, and then decreased to the peak at the 14th day (P 0.05) and decreased at 28 days. Conclusion autophagy is activated in alveolar macrophages of silicosis rats, and alveolar macrophage autophagy is involved in the pathological process of silicosis in rats.
【作者单位】：河北联合大学基础医学院组织学胚胎学系;河北联合大学冀唐学院;唐山市工人医院神经外科;河北省煤矿卫生与安全实验室;唐山市慢性病基础与临床实验室;
【基金】：河北省科技支撑重点专项资助项目(09276191D) 河北省重点职业病防治技术研究资助项目(13277709D)
【分类号】：R135.2

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