| 顾超,陶峰,曹林峰,李娜,应樱,张齐,马肖龙.II型肺泡上皮细胞凋亡在COPD患者肺损伤中的作用及机制研究[J].浙江中西医结合杂志,2022,32(8): |
| II型肺泡上皮细胞凋亡在COPD患者肺损伤中的作用及机制研究 |
| The effects of type II alveolar epithelial cell apoptosis on lung injury in patients with chronic obstructive pulmonary disease |
| 投稿时间：2021-10-03 修订日期：2022-07-18 |
| DOI： |
| 中文关键词: II型肺泡上皮细胞 慢性阻塞性肺疾病 细胞凋亡 吸烟 叉头框蛋白O3a |
| 英文关键词:Type II alveolar epithelial cells Chronic obstructive pulmonary disease Cell apoptosis Smoking FoxO3a |
| 基金项目:浙江省嘉兴市科技计划项目(No.2017AY33010)；嘉兴市第一医院院级课题重点项目(No.2020YJZD017) |
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| 中文摘要: |
| 目的 分析Ⅱ型肺泡上皮细胞（AECⅡ）凋亡在慢性阻塞性肺疾病（COPD）患者肺损伤中的作用及其机制。方法 筛选2020年1月至2020年12月浙江省嘉兴市第一医院手术治疗的非小细胞肺癌（NSCLC）患者30例入组，分为NSCLC不伴COPD且无吸烟史的患者（A组）、NSCLC不伴COPD但有吸烟史的患者（B组）及NSCLC伴COPD且有吸烟史的患者（C组），每组10例，常规测定肺功能。分别采用HE染色观察肺组织病理学变化，Real-time PCR和免疫组化检测肺泡表面活性蛋白A（SPA）和肺泡表面活性蛋白C（SPC）mRNA及蛋白表达水平，TUNEL/SPC双重免疫荧光染色检测AECⅡ凋亡情况，Western blot检测p53、p21及叉头框蛋白O3a（FoxO3a）蛋白表达变化。结果 C组患者肺功能符合COPD诊断标准，即FEV1/FVC为（64.90±3.00）%，肺组织呈现明显的肺气肿样病理学改变。分别与A、B组比较，C组MLI[（194.01±7.01） μm比（36.34±4.09） μm、（39.49±4.01） μm]；MAA[（21707.72±2306.53） μm2比（3352.49±391.44） μm2、（3493.38±404.29） μm2]明显升高（P均<0.05）；MAN[（52.46±4.68） mm2比（272.50±23.42） mm2、（255.77±32.06） mm2]明显降低（P<0.05）。Real-time PCR和免疫组化结果显示，分别与A、B组比较，C组SPA mRNA[（0.71±0.08）比（1.00±0.02）、（0.94±0.04）]，SPC mRNA[（0.72±0.08）比（1.00±0.02）、（0.96±0.03）]明显减少（P<0.05）。TUNEL/SPC双重免疫荧光染色结果表明，C组的AECⅡ凋亡百分比为（12.30±2.21）%，显著高于A组（5.60±1.71）%和B组（6.00±1.83）%，差异有统计学意义（P<0.05）。Western blot结果表明，分别与A组和B组比较，C组患者肺组织p53和p21蛋白表达量明显增加（P<0.05）、FoxO3a蛋白表达量显著降低（P<0.05）。结论 AECⅡ凋亡在COPD患者肺损伤中起到重要作用，其机制可能与FoxO3a蛋白表达下调及p53、p21蛋白表达上调相关。 |
| 英文摘要: |
| Objective To analyze the role and mechanism of type II alveolar epithelial cell (AECII) apoptosis on lung injury in patients with chronic obstructive pulmonary disease (COPD). Methods Thirty patients with non-small cell lung cancer (NSCLC) who were surgically treated in the First Hospital of Jiaxing, Zhejiang Province from January 2020 to December 2020 were selected and divided into three groups: NSCLC patients without COPD and smoking history (group A, n=10), NSCLC patients without COPD but with smoking history (group B, n=10) and NSCLC patients with COPD and smoking history (group C, n=10). Pulmonary function was measured routinely. The pathological changes of lung tissue were observed by HE staining. Real-time PCR and immunohistochemistry were used to detect the mRNA and protein expression levels of surfactant protein A (SPA) and surfactant protein C (SPC). TUNEL/SPC double immunofluorescence staining was used to detect the apoptosis of AECII. Western blot was used to detect the protein expression changes of p53, p21 and forkhead box protein O3a (FoxO3a). Results The lung function of patients in group C met the diagnostic criteria of COPD, that is (64.90±3.00)% of FEV1/FVC, and the lung tissue showed obvious emphysema-like pathological changes. Compared with group A and group B, the MLI in group C was (194.01±7.01) μm compared with (36.34±4.09) μm and (39.49±4.01) μm; the MAA (21707.72±2306.53) μm2 in group C was significantly higher than that of (3352.49±391.44) μm2 and (3493.38±404.29) μm2 (all P < 0.05); the MAN in group C (52.46±4.68) mm2 was significantly lower than that of (272.50±23.42) mm2 and (255.77±32.06) mm2 (P<0.05). Real-time PCR and immunohistochemistry showed that the expression levels of SPA mRNA (0.71±0.08) and SPC mRNA(0.72±0.08) in group C were significantly lower than those in group A [(1.00±0.02) and (1.00±0.02)] and group B [(0.96±0.04) and (0.96±0.03)] (P<0.05), respectively. Meanwhile, the expressions of SPA and SPC proteins in group C were significantly decreased. The TUNEL/SPC double immunofluorescence staining results showed that the percentage of AECII apoptosis in group C was (12.30±2.21)%, significantly higher than those in group A (5.60±1.71)% and group B (6.00±1.83)% (P<0.05). Western blot results showed that compared with group A and group B, the expressions of p53 and p21 proteins in the lung tissue of group C were significantly increased (P<0.05), and the expression of FoxO3a protein was significantly decreased (P<0.05). Conclusion AECII apoptosis plays an important role in lung injury in patients with COPD, and its mechanism may be related to the down-regulation of FoxO3a protein expression and up-regulation of p53 and p21 proteins. |
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