慢性阻塞性肺疾病患者肺组织中自噬相关基因和蛋白的表达

发布时间：2018-03-02 03:13

本文关键词： 慢性阻塞性肺疾病人肺组织自噬　出处：《上海医学》2017年02期 　论文类型：期刊论文

【摘要】：目的探讨慢性阻塞性肺疾病(COPD)患者肺组织标本中自噬相关基因和蛋白的表达情况。方法标本取自45例因肺部占位性病变行肺叶切除术患者的肺组织,根据患者有无COPD,分为非COPD组(对照组,15例)和COPD组(30例);再根据COPD患者的肺功能,将COPD组分为轻度COPD组(12例)、中度COPD组(12例)、重度COPD组(6例)。采用免疫组织化学染色观察肺组织中自噬蛋白微管相关蛋白1轻链3(LC3)B的表达;采用Western印迹法和实时定量PCR检测肺组织标本中自噬相关基因Atg5、酵母自噬相关基因Atg6同系物(Beclin1),以及自噬相关基因Atg5、Beclin1、LC3B的蛋白表达。结果非COPD组的肺组织结构清晰,未见支气管、肺泡结构破坏,炎性细胞浸润等。COPD组的肺组织结构破坏明显,肺气肿形成,肺实质有肺泡管、肺泡腔扩大,上皮细胞脱落、坏死等。LC3B蛋白主要表达于肺上皮细胞细胞质中,可见清晰的棕黄色颗粒。与非COPD组相比,COPD组肺上皮细胞细胞质中棕黄色颗粒增多,染色强度增加。轻度COPD组、中度COPD组和重度COPD组的Beclin1 mRNA均显著高于非COPD组(P值分别0.01、0.05),轻度COPD组和中度COPD组的Atg5 mRNA均显著高于非COPD组(P值分别0.01、0.05),重度COPD组与非COPD组间Atg5 mRNA的差异无统计学意义(P0.05)。中度COPD组的Beclin1 mRNA和Atg5 mRNA分别显著高于轻度COPD组和重度COPD组(P值均0.01),轻度COPD组的Beclin1 mRNA和Atg5 mRNA分别显著高于重度COPD组(P值均0.05)。轻度COPD组和中度COPD组的Atg5蛋白表达量均显著高于非COPD组(P值分别0.01、0.05),重度COPD组与非COPD组间Atg5蛋白表达量的差异无统计学意义(P0.05);轻度COPD组、中度COPD组和重度COPD组的Beclin1蛋白和LC3B蛋白表达量均显著高于非COPD组(P值分别0.01、0.05)。中度COPD组的Beclin1、Atg5、LC3B蛋白表达量分别显著高于轻度COPD组和重度COPD组(P值分别0.05、0.01),轻度COPD组的Beclin1、Atg5、LC3B蛋白表达量分别显著高于重度COPD组(P值均0.05)。结论在COPD的进展过程中自噬相关基因和蛋白表达增多,提示可能与疾病的发展有关。
[Abstract]:Objective to investigate the expression of autophagy related genes and proteins in lung tissue of patients with chronic obstructive pulmonary disease (COPD). According to whether the patients had copd, they were divided into non COPD group (control group, 15 cases) and COPD group (30 cases), and then according to the lung function of COPD patients, The COPD group was divided into mild COPD group (n = 12), moderate COPD group (n = 12) and severe COPD group (n = 6). The expression of autophagy related gene Atg5, yeast autophagy associated gene Atg6 congener Atg6, and autophagy associated gene Atg5, Beclin1 and LC3B were detected by Western blot and real-time quantitative PCR. Results the lung tissue structure of non-#en3# group was clear and no bronchus was found. Destruction of alveolar structure, infiltration of inflammatory cells, obvious destruction of lung tissue structure, formation of emphysema, pulmonary parenchyma with alveolar duct, enlargement of alveolar cavity, exfoliation and necrosis of epithelial cells, etc., etc. The protein of .LC3B was mainly expressed in the cytoplasm of pulmonary epithelial cells. Compared with the non COPD group, the brown granules in the cytoplasm of pulmonary epithelial cells increased and the staining intensity increased in the COPD group. The Beclin1 mRNA of moderate COPD group and severe COPD group were significantly higher than that of non-#en4# group (P < 0.01). Atg5 mRNA of mild COPD group and moderate COPD group were significantly higher than that of non-#en9# group. There was no significant difference in Atg5 mRNA between severe COPD group and non-#en11# group. Beclin1 mRNA and Atg5 mRNA in moderate COPD group were significantly higher than those in mild COPD group and severe COPD group, P values of Beclin1 mRNA and Atg5 mRNA in mild COPD group were significantly higher than those in severe COPD group, respectively. The expression of Atg5 protein in mild COPD group and moderate COPD group was significantly higher than that in severe COPD group. There was no significant difference in the expression of Atg5 protein between the severe COPD group and the non-#en2# group (P 0.05), while in the mild COPD group, there was no significant difference in the expression of Atg5 protein between the severe COPD group and the non COPD group. The expression of Beclin1 protein and LC3B protein in moderate COPD group and severe COPD group were significantly higher than those in non-#en4# group (P = 0.01, P = 0.05). The expression of Beclin1 COPD and LC3B protein in moderate COPD group was significantly higher than that in mild COPD group and severe COPD group (P = 0.05), and that in mild COPD group was significantly higher than that in mild COPD group and severe COPD group, respectively. Protein expression levels were significantly higher in severe COPD group than in severe COPD group (P < 0.05). Conclusion the expression of autophagy related genes and proteins increased during the progression of COPD. This may be related to the development of the disease.
【作者单位】：上海交通大学医学院附属新华医院呼吸科;
【基金】：上海市自然科学基金项目资助(14ZR1426800)
【分类号】：R563.9

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