本文關(guān)鍵詞： 慢性阻塞性肺疾病 人 肺組織 自噬　出處：《上海醫(yī)學(xué)》2017年02期 　論文類(lèi)型：期刊論文

【摘要】：目的探討慢性阻塞性肺疾病(COPD)患者肺組織標(biāo)本中自噬相關(guān)基因和蛋白的表達(dá)情況。方法標(biāo)本取自45例因肺部占位性病變行肺葉切除術(shù)患者的肺組織,根據(jù)患者有無(wú)COPD,分為非COPD組(對(duì)照組,15例)和COPD組(30例);再根據(jù)COPD患者的肺功能,將COPD組分為輕度COPD組(12例)、中度COPD組(12例)、重度COPD組(6例)。采用免疫組織化學(xué)染色觀察肺組織中自噬蛋白微管相關(guān)蛋白1輕鏈3(LC3)B的表達(dá);采用Western印跡法和實(shí)時(shí)定量PCR檢測(cè)肺組織標(biāo)本中自噬相關(guān)基因Atg5、酵母自噬相關(guān)基因Atg6同系物(Beclin1),以及自噬相關(guān)基因Atg5、Beclin1、LC3B的蛋白表達(dá)。結(jié)果非COPD組的肺組織結(jié)構(gòu)清晰,未見(jiàn)支氣管、肺泡結(jié)構(gòu)破壞,炎性細(xì)胞浸潤(rùn)等。COPD組的肺組織結(jié)構(gòu)破壞明顯,肺氣腫形成,肺實(shí)質(zhì)有肺泡管、肺泡腔擴(kuò)大,上皮細(xì)胞脫落、壞死等。LC3B蛋白主要表達(dá)于肺上皮細(xì)胞細(xì)胞質(zhì)中,可見(jiàn)清晰的棕黃色顆粒。與非COPD組相比,COPD組肺上皮細(xì)胞細(xì)胞質(zhì)中棕黃色顆粒增多,染色強(qiáng)度增加。輕度COPD組、中度COPD組和重度COPD組的Beclin1 mRNA均顯著高于非COPD組(P值分別0.01、0.05),輕度COPD組和中度COPD組的Atg5 mRNA均顯著高于非COPD組(P值分別0.01、0.05),重度COPD組與非COPD組間Atg5 mRNA的差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。中度COPD組的Beclin1 mRNA和Atg5 mRNA分別顯著高于輕度COPD組和重度COPD組(P值均0.01),輕度COPD組的Beclin1 mRNA和Atg5 mRNA分別顯著高于重度COPD組(P值均0.05)。輕度COPD組和中度COPD組的Atg5蛋白表達(dá)量均顯著高于非COPD組(P值分別0.01、0.05),重度COPD組與非COPD組間Atg5蛋白表達(dá)量的差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);輕度COPD組、中度COPD組和重度COPD組的Beclin1蛋白和LC3B蛋白表達(dá)量均顯著高于非COPD組(P值分別0.01、0.05)。中度COPD組的Beclin1、Atg5、LC3B蛋白表達(dá)量分別顯著高于輕度COPD組和重度COPD組(P值分別0.05、0.01),輕度COPD組的Beclin1、Atg5、LC3B蛋白表達(dá)量分別顯著高于重度COPD組(P值均0.05)。結(jié)論在COPD的進(jìn)展過(guò)程中自噬相關(guān)基因和蛋白表達(dá)增多,提示可能與疾病的發(fā)展有關(guān)。
[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.
【作者單位】： 上海交通大學(xué)醫(yī)學(xué)院附屬新華醫(yī)院呼吸科;
【基金】：上海市自然科學(xué)基金項(xiàng)目資助(14ZR1426800)
【分類(lèi)號(hào)】：R563.9

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