| 白莉敏,吕 行,任引刚,祝松涛,杨 璐.miR-22-3p通过激活PTEN/PI3K/AKT/NF-κB信号通路加重慢性阻塞性肺疾病[J].,2023,(10):1835-1842 |
| miR-22-3p通过激活PTEN/PI3K/AKT/NF-κB信号通路加重慢性阻塞性肺疾病 |
| miR-22-3p Aggravates Chronic Obstructive Pulmonary Disease through Activating PTEN/PI3K/AKT/NF-κB Signaling Pathway |
| 投稿时间：2022-12-23 修订日期：2023-01-31 |
| DOI：10.13241/j.cnki.pmb.2023.10.006 |
| 中文关键词: 慢性阻塞性肺疾病 miR-22-3p 炎症 磷酸酶与张力蛋白同源物 |
| 英文关键词: Chronic obstructive pulmonary disease MiR-22-3p Inflammation Phosphatase and tensin homologues |
| 基金项目:陕西省卫生健康委科研基金项目（2021A006） |
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| 中文摘要: |
| 摘要 目的：探索miR-22-3p对慢性阻塞性肺疾病（COPD）的影响及作用机制。方法：将7周龄（体重250~280 g）雄性SD大鼠随机分为5组（n=12）：对照组（Con组）、COPD组、COPD+NC-agomir组、COPD+miR-22-3p-agomir组、COPD+NC-antagomir组和COPD+miR-22-3p-antagomir组。Con组大鼠为正常饲养的大鼠，其他组大鼠均通过香烟烟雾（CS）和脂多糖（LPS）诱导COPD动物模型。在CS暴露当天，Con组和COPD组大鼠鼻腔内注射50 μL生理盐水，COPD+NC-agomir组、COPD+miR-22-3p-agomir组、COPD+NC-antagomir组和COPD+miR-22-3p-antagomir组大鼠分别鼻腔内注射50 μL 10 nmoL的NC-agomir、miR-22-3p-agomir、NC-antagomir和miR-22-3p-antagomir，每2周注射1次，共注射6次。CS暴露90 d后，检测各组大鼠的最大自主分钟通气量（MVV）、0.3 s用力呼气量（FEV0.3）、用力肺活量（FVC）和最大呼气峰流速（PEF）。通过ELISA法测定支气管肺泡灌洗液（BALF）中白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）和巨噬细胞炎性蛋白-2（MIP-2）水平。通过苏木精-伊红（HE）染色评价肺组织损伤。根据试剂盒说明检测肺组织丙二醛（MDA）和超氧化物歧化酶（SOD）水平。通过RT-PCR检测肺组织miR-22-3p、磷酸酶与张力蛋白同源物（PTEN）、诱导型一氧化氮合酶（iNOS）、CD86、CD206和精氨酸酶1（ARG1）mRNA水平。通过Western blot检测肺组织PTEN、磷脂酰肌醇3-激酶（PI3K）、p-PI3K、蛋白激酶B（AKT）、p-AKT、核因子-κB（NF-κB） p65和p-NF-κB p65的蛋白表达水平。结果：与COPD组和COPD+NC-antagomir组相比，COPD+miR-22-3p-antagomir组大鼠的MVV、FEV0.3/FVC和PEF均升高（P<0.05）；肺泡出血、肺泡壁断裂、肺泡间隔水肿和炎性细胞浸润等病变减轻；BALF中的IL-1β、TNF-α和MIP-2水平均降低（P<0.05）；肺组织SOD水平升高，MDA水平降低（P<0.05）；肺组织iNOS和CD86 mRNA相对表达量降低，CD206和ARG1 mRNA相对表达量升高（P<0.05）；肺组织miR-22-3p相对表达量降低，PTEN mRNA和蛋白相对表达量升高（P<0.05）；肺组织p-PI3K/PI3K、p-AKT/AKT和p-NF-κB p65/NF-κB p65的蛋白相对表达量降低（P<0.05）。结论：miR-22-3p在COPD大鼠肺组织中上调，可能通过激活PTEN/PI3K/AKT/NF-κB信号通路，促进肺组织炎症反应和氧化应激，加重大鼠肺功能障碍和肺组织结构损伤。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the effect and mechanism of miR-22-3p on chronic obstructive pulmonary disease (COPD). Methods: Male SD rats aged 7 weeks (weight 250 g to 280 g) were randomly divided into 5 groups (n=12): Control group (Con), COPD group, COPD+NC-agomir group, COPD+miR-22-3p-agomir group, COPD+NC-antagomir group and COPD+miR-22-3p-antagomir group. The rats in the Con group were normally fed, and the other groups were induced by cigarette smoke (CS) and lipopolysaccharide (LPS). On the day of CS exposure, 50 μL saline was injected intranasally in Con group and COPD group rats. And the rats in COPD+NC agomir group, COPD+miR-22-3p-agomir group, COPD+NC antagomir group and COPD+miR-22-3p-antagomir group were given intranasal injection of 50 μL 10 nmol of NC agomir, miR-22-3p-agomir, NC antagomir and miR-22-3p-antagomir, respectively. It was injected once every 2 weeks for a total of 6 times. After 90 days of CS exposure, the maximum autonomous ventilation volume per minute (MVV), 0.3 s forced expiratory volume (FEV0.3), forced vital capacity (FVC) and peak expiratory flow (PEF) were measured. The levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and macrophage inflammatory protein-2 (MIP-2) in bronchoalveolar lavage fluid (BALF) were measured by ELISA method. Lung tissue injury was evaluated by hematoxylin-eosin (HE) staining. The levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in lung tissue were measured according to the instructions of the kit. The mRNA levels of miR-22-3p, phosphatase and tensin homologue (PTEN), inducible nitric oxide synthase (iNOS), CD86, CD206 and argininase 1 (ARG1) in lung tissue were measured by RT-PCR. The protein levels of PTEN, phosphatidylinositol 3-kinase (PI3K), p-PI3K, protein kinase B (AKT), p-AKT, nuclear factor-kappa B (NF-κB) p65 and p-NF-κB p65 in lung tissue were detected by Western blot. Results: Compared with COPD group and COPD+NC-antagomir group, the MVV, FEV0.3/FVC and PEF of COPD+miR-22-3p-antagomir group increased (P<0.05), alveolar hemorrhage, rupture of alveolar wall, edema of alveolar septum and inflammatory cell infiltration were alleviated, the levels of IL-1β, TNF-α and MIP-2 in BALF decreased(P<0.05), the level of SOD in lung tissue increased and the level of MDA decreased(P<0.05), the relative expression of iNOS and CD86 mRNA in lung tissue decreased(P<0.05), while the relative expression of CD206 and ARG1mRNA increased(P<0.05), the relative expression levels of miR-22-3p in lung tissues decreased, while the relative expression levels of PTEN mRNA and protein increased (P<0.05), the relative expression of p-PI3K/PI3K, p-AKT/AKT and p-NF-κB p65/NF-κB p65 decreased in lung tissue(P<0.05). Conclusion: miR-22-3p is up-regulated in the lung tissues of COPD rats, which may promotes the inflammation and oxidative stress in the lung and aggravates lung dysfunction and lung injury through activating PTEN/PI3K/AKT/NF-κB signaling pathway. |
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