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[1]謝叢意 姚秀娟 蘭玉珠 謝寶松.大鼠間質(zhì)性肺炎Ⅱ型肺泡上皮細(xì)胞自噬活性的變化及其機(jī)制研究[J].福建醫(yī)藥雜志,2019,41(03):116-120.
　XIE Congyi,YAO Xiujuan,LAN Yuzhu,et al.Changes of autophagy activity and its mechanism in type Ⅱ alveolar epithelial cells of rats with interstitial pneumonia[J].FUJIAN MEDICAL JOURNAL,2019,41(03):116-120.
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大鼠間質(zhì)性肺炎Ⅱ型肺泡上皮細(xì)胞自噬活性的變化及其機(jī)制研究()
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《福建醫(yī)藥雜志》[ISSN:1002-2600/CN:35-1071/R]

卷:
41
期數(shù):
2019年03期
頁(yè)碼:
116-120
欄目:
基礎(chǔ)研究
出版日期:
2019-02-20

文章信息/Info

Title:
Changes of autophagy activity and its mechanism in type Ⅱ alveolar epithelial cells of rats with interstitial pneumonia
文章編號(hào):
1002-2600(2019)03-0116-05
作者:
謝叢意 姚秀娟1 蘭玉珠2 謝寶松1; 3
福建中醫(yī)藥大學(xué)(福州 350122)
Author(s):
XIE Congyi; YAO Xiujuan; LAN Yuzhu; XIE Baosong.
Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350122, China
關(guān)鍵詞:
自噬;  間質(zhì)性肺炎;  肺纖維化;  博來(lái)霉素
Keywords:
autophagy;  interstitial pneumonia;  pulmonary fibrosis;  bleomycin
分類號(hào):
R563.1+3
文獻(xiàn)標(biāo)志碼:
A
摘要:
目的 探討大鼠間質(zhì)性肺炎Ⅱ型肺泡上皮細(xì)胞自噬活性的變化及其機(jī)制。方法 將16只Wistar雄性大鼠隨機(jī)數(shù)字分為對(duì)照組(A組)和實(shí)驗(yàn)組(B組),每組8只。實(shí)驗(yàn)組霧化吸入博來(lái)霉素,對(duì)照組霧化吸入等量生理鹽水。所有大鼠于28 d處死,收集各組肺組織標(biāo)本。蘇木精―伊紅染色(HE)觀察肺組織病理改變; Masson三色染色法觀察肺組織膠原纖維生成情況; 采用免疫黏附法分離Ⅱ型肺泡上皮細(xì)胞后,采用蛋白質(zhì)免疫印跡法(Western blot)檢測(cè)-平滑肌肌動(dòng)蛋白(α-SMA)、泛素結(jié)合蛋白p62、自噬微管相關(guān)蛋白輕鏈3(LC3Ⅰ/Ⅱ)、磷酸化的蛋白激酶B(p-Akt)、哺乳動(dòng)物雷帕霉素靶蛋白(mTOR)及其磷酸化形式p-mTOR的表達(dá)變化。結(jié)果 1)HE染色顯示,A組大鼠肺組織呈正常表現(xiàn),未見(jiàn)肺泡炎及纖維化改變; B組肺實(shí)質(zhì)呈大片融合實(shí)變,其間有少量巨噬細(xì)胞、中性粒細(xì)胞等炎性細(xì)胞浸潤(rùn),成纖維細(xì)胞增多,肺泡間隔明顯增厚,可見(jiàn)膠原沉積。2)Masson染色顯示,A和B組藍(lán)色區(qū)域面積分別為(12.12±1.61)mm2、(23.73±4.20)mm2,B組藍(lán)色面積大于A組,差異有統(tǒng)計(jì)學(xué)意義(P<0.05)); A組藍(lán)色區(qū)域主要集中在氣管或血管周圍,肺間質(zhì)只有少量散在的點(diǎn)狀著色,B組藍(lán)色區(qū)域除氣管和血管周圍外,肺間質(zhì)也可見(jiàn)大量藍(lán)色著色區(qū),呈片絮狀。3)Western blot結(jié)果顯示,B組α-SMA蛋白相對(duì)表達(dá)量高于A組; B組p62蛋白相對(duì)表達(dá)量高于A組; B組LC3Ⅱ/Ⅰ比值低于A組; B組p-Akt和p-mTOR相對(duì)表達(dá)量均高于A組,差異均有統(tǒng)計(jì)學(xué)意義(均P<0.05); A組和B組mTOR蛋白相對(duì)表達(dá)量差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.05)。結(jié)論 大鼠間質(zhì)性肺炎Ⅱ型肺泡上皮細(xì)胞出現(xiàn)細(xì)胞自噬活性抑制,Akt/mTOR信號(hào)通路激活是其可能的機(jī)制之一。
Abstract:
Objective To investigate the changes of autophagy activity and its mechanism in type Ⅱ alveolar epithelial cells of rats with interstitial pneumonia.Methods Sixteen male rats were randomly and averagely assigned into control group(Group A)and experimental group(Group B).Experimental group received bleomycin inhalation, while the control group received saline inhalation.Rats were sacrificed on the 28h days and lung tissue were collected.Hematoxylin-eosin staining(HE)was used to observe the pathological changes of lung tissue.Collagen fibers were evaluated by Masson staining.Type Ⅱ alveolar epithelia were obtained by immune adhesion, and the expression of α-SMA and autophagy markers(p62 and LC3Ⅰ/Ⅱ)as well as signaling pathway proteins(p-mTOR/mTOR and p-Akt)of type Ⅱ alveolar cells were assessed with Western blot.Results 1)Lung tissue morphology of group A was normal.In group B, large consolidation of lung parenchyma with mild infiltration of inflammatory cells(macrophages and neutrophils)and fibroblasts were found, and the alveolar interval was thickened with collagen deposition.2)Masson staining showed that the blue area of Group B was significantly larger than Group A [(23.73±4.20)mm2 vs(12.12±1.61)mm2, P<0.05].The stained area of Group A was concentrated around trachea and vessels, while the stained area of Group B was further detected in the lung mesenchyme, presented as flake form.3)Western blot showed that the expression of α-SMA of experimental group was higher than control group(P<0.05).The p62 levels of Group B were significantly higher than Group A(P<0.05).LC3 Ⅱ/Ⅰ ratio of Group B was lower than Group A(P<0.05).The expression of p-Akt and p-mTOR of experimental group was higher than control group(P<0.05).There was no significant difference in the expression of mTOR between the two groups(P>0.05).Conclusion Autophagy activity is inhibited in type Ⅱ alveolar epithelial cells of rat with interstitial pneumonia,possibly due to the activation of Akt/mTOR signaling pathway.

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備注/Memo

備注/Memo:
1 福建醫(yī)科大學(xué)省立臨床醫(yī)學(xué)院呼吸與危重癥醫(yī)學(xué)科; 2 福建醫(yī)科大學(xué); 3 通信作者,Email:[email protected]
更新日期/Last Update: 2019-02-20