Gao Hai-Xiang, Su Yan, Zhang Ai-Li, Xu Jin-Wei, Fu Qian, Yan Li
a Department of Respiratory Medicine , Hebei General Hospital , Shijiazhuang , China.
Exp Lung Res. 2019 Feb-Mar;45(1-2):1-12. doi: 10.1080/01902148.2018.1563925. Epub 2019 Apr 29.
To study the role of miR-34c-5p targeting in affecting the progression of chronic obstructive pulmonary disease (COPD). The dual-luciferase reporter gene assay was applied to verify the targeting relationship of miR-34c-5p and . The rats were randomly assigned into Control, COPD, COPD + empty plasmids, COPD + agomir, COPD + CCL22 shRNA and COPD + agomir + CCL22 groups. COPD model was built by using cigarette smoke exposure and LPS instillation. After 28 days, the pulmonary function was examined. ELISA method was used to detect TNF-α and IL-8 levels in bronchoalveolar lavage fluid (BALF), HE staining and Masson staining to observe the pathomorphological changes of lung tissues, qRT-PCR and/or Western blot to determine miR-34c-5p and CCL22 levels, and immunohistochemical staining to measure the expression of MMP-9 and TIMP-1. MiR-34c-5p could target to down-regulate its expression. Both miR-34c-5p agomir and CCL22 shRNA could reduce breathing frequency (f), airway resistance (RI), and the levels of IL-8 and TNF-α in BALF of COPD rats with increased Cydn (dynamic lung compliance) and PIF (peak inspiratory flow). Besides, the inflammatory cell infiltration, rupture of partial alveolus, enlarged alveolar cavity, and increased deposition of collagen fibers were observed in COPD rat tissues, with rise in mean linear intercept (MLI) and reduction in mean alveolar number (MAN), which could be reversed by miR-34c-5p agomir or CCL22 shRNA. MiR-34c-5p may promote inflammation response and maintain the protease-antiprotease balance via targeting , which provides scientific basis for the clinical treatment of COPD.
研究miR-34c-5p靶向作用在影响慢性阻塞性肺疾病（COPD）进展中的作用。应用双荧光素酶报告基因检测法验证miR-34c-5p与……的靶向关系。将大鼠随机分为对照组、COPD组、COPD+空质粒组、COPD+激动剂组、COPD+CCL22短发夹RNA组和COPD+激动剂+CCL22组。采用香烟烟雾暴露和脂多糖灌胃法建立COPD模型。28天后，检测肺功能。采用酶联免疫吸附测定（ELISA）法检测支气管肺泡灌洗液（BALF）中肿瘤坏死因子-α（TNF-α）和白细胞介素-8（IL-8）水平，苏木精-伊红（HE）染色和Masson染色观察肺组织病理形态学变化，实时定量聚合酶链反应（qRT-PCR）和/或蛋白质免疫印迹法（Western blot）检测miR-34c-5p和CCL22水平，免疫组织化学染色检测基质金属蛋白酶-9（MMP-9）和金属蛋白酶组织抑制因子-1（TIMP-1）的表达。miR-34c-5p可靶向……以下调其表达。miR-34c-5p激动剂和CCL22短发夹RNA均可降低COPD大鼠的呼吸频率（f）、气道阻力（RI）以及BALF中IL-8和TNF-α水平，同时增加动态肺顺应性（Cydn）和吸气峰流速（PIF）。此外，在COPD大鼠组织中观察到炎性细胞浸润、部分肺泡破裂、肺泡腔增大以及胶原纤维沉积增加，平均线性截距（MLI）升高，平均肺泡数（MAN）减少，而miR-34c-5p激动剂或CCL22短发夹RNA可使其逆转。miR-34c-5p可能通过靶向……促进炎症反应并维持蛋白酶-抗蛋白酶平衡，为COPD的临床治疗提供科学依据。
