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基于呼吸道菌群特征对慢性阻塞性肺疾病频繁急性加重者病理机制的研究

Investigation on the Pathological Mechanism of Frequent Exacerbators With Chronic Obstructive Pulmonary Disease Based on the Characteristics of Respiratory Flora.

作者信息

Ke Li, Chen Luo, Yaling Yuan, Can Gao, Jun Lin, Chuan Zhang

机构信息

Department of Laboratory Medicine, Chongqing the Seventh People's Hospital, Chongqing, China.

Department of Respiratory and Critical Care, Chongqing the Seventh People's Hospital, Chongqing, China.

出版信息

Front Med (Lausanne). 2022 Jan 20;8:816802. doi: 10.3389/fmed.2021.816802. eCollection 2021.

Abstract

Chronic obstructive pulmonary disease (COPD) is a common obstructive respiratory disease characterized by persistent respiratory symptoms and limited airflow due to airway obstruction. The present study investigates the distribution characteristics of respiratory tract flora in both frequent and infrequent exacerbators of COPD. The 16S sequencing technique was adopted to differentiate the inherent differences of respiratory tract flora between frequent exacerbators and infrequent exacerbators. Additionally, cell counting kit 8 (CCK8), lactate dehydrogenase (LDH) test, flow cytometry, enzyme-linked immunosorbent assay (ELISA), and western blot were carried out in human bronchial epithelial cells cultured and the regulatory effects of differential flora were verified. The results revealed that the observed species index, Chao1 index, and the ACE estimator of COPD frequent exacerbators were markedly higher than those of COPD infrequent exacerbators. The top five strains of COPD frequent exacerbators included g_Streptococcus (15.565%), g_Prevotella (10.683%), g_Veillonella (6.980%), g_Haemophilus (5.601%), and g_Neisseria (4.631%). Veillonella parvula generated obvious cytotoxicity and substantially reduced the activity of human bronchial epithelial cells ( < 0.01). Furthermore, the results of flow cytometry indicated that the proportion of human bronchial epithelial cells in both the S phase and G2 phase decreased following Veillonella parvula treatment indicated that Veillonella parvula inhibited cell proliferation. Meanwhile, being treated using Veillonella parvula, the expressions of interleukin-1 (IL-1), IL-6, Tumor Necrosis Factor α (TNF-α), and p-nuclear factor kappa B (NF-κB) of the cells were increased markedly ( < 0.01). Taken together, the current research demonstrated that the relative abundance of Veillonella in COPD frequent exacerbators was higher than that of infrequent exacerbators. Veillonella parvula activated the inflammatory pathway, ultimately destroyed the cell viability, and greatly impaired the activity of human bronchial epithelial cells, thereby inhibiting cell proliferation.

摘要

慢性阻塞性肺疾病(COPD)是一种常见的阻塞性呼吸道疾病,其特征为持续的呼吸道症状以及由于气道阻塞导致的气流受限。本研究调查了COPD频繁加重者和不频繁加重者呼吸道菌群的分布特征。采用16S测序技术来区分频繁加重者和不频繁加重者呼吸道菌群的内在差异。此外,在培养的人支气管上皮细胞中进行了细胞计数试剂盒8(CCK8)、乳酸脱氢酶(LDH)检测、流式细胞术、酶联免疫吸附测定(ELISA)和蛋白质免疫印迹法,并验证了差异菌群的调节作用。结果显示,COPD频繁加重者的观测物种指数、Chao1指数和ACE估计值显著高于COPD不频繁加重者。COPD频繁加重者的前五种菌株包括g_链球菌(15.565%)、g_普雷沃菌(10.683%)、g_韦荣球菌(6.980%)、g_嗜血杆菌(5.601%)和g_奈瑟菌(4.631%)。小韦荣球菌产生明显的细胞毒性,并显著降低人支气管上皮细胞的活性(<0.01)。此外,流式细胞术结果表明,小韦荣球菌处理后人支气管上皮细胞在S期和G2期的比例均下降,表明小韦荣球菌抑制细胞增殖。同时,用小韦荣球菌处理后,细胞的白细胞介素-1(IL-1)、IL-6、肿瘤坏死因子α(TNF-α)和磷酸化核因子κB(NF-κB)的表达显著增加(<0.01)。综上所述,当前研究表明,COPD频繁加重者中小韦荣球菌的相对丰度高于不频繁加重者。小韦荣球菌激活炎症途径,最终破坏细胞活力,并极大损害人支气管上皮细胞的活性,从而抑制细胞增殖。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee1d/8811034/9f8724037fa8/fmed-08-816802-g0001.jpg

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