Divisione di Pneumologia e Laboratorio di Citoimmunopatologia dell'Apparato Cardio Respiratorio, Istituti Clinici Scientifici Maugeri, IRCCS, Novara, Veruno, Italy.
Dipartimento di Scienze Mediche, Università di Torino, Turin, Italy.
Can Respir J. 2020 Aug 8;2020:1524716. doi: 10.1155/2020/1524716. eCollection 2020.
Chronic obstructive pulmonary disease (COPD) is due to structural changes and narrowing of small airways and parenchymal destruction (loss of the alveolar attachment as a result of pulmonary emphysema), which all lead to airflow limitation. Extracorporeal shock waves (ESW) increase cell proliferation and differentiation of connective tissue fibroblasts. To date no studies are available on ESW treatment of human bronchial fibroblasts and epithelial cells from COPD and control subjects. We obtained primary bronchial fibroblasts from bronchial biopsies of 3 patients with mild/moderate COPD and 3 control smokers with normal lung function. 16HBE cells were also studied. Cells were treated with a piezoelectric shock wave generator at low energy (0.3 mJ/mm, 500 pulses). After treatment, viability was evaluated and cells were recultured and followed up for 4, 24, 48, and 72 h. Cell growth (WST-1 test) was assessed, and proliferation markers were analyzed by qRT-PCR in cell lysates and by ELISA tests in cell supernatants and cell lysates. After ESW treatment, we observed a significant increase of cell proliferation in all cell types. C-Kit (CD117) mRNA was significantly increased in 16HBE cells at 4 h. Protein levels were significantly increased for c-Kit (CD117) at 4 h in 16HBE ( < 0.0001) and at 24 h in COPD-fibroblasts ( = 0.037); for PCNA at 4 h in 16HBE ( = 0.046); for Thy1 (CD90) at 24 and 72 h in CS-fibroblasts ( = 0.031 and = 0.041); for TGF1 at 72 h in CS-fibroblasts ( = 0.038); for procollagen-1 at 4 h in COPD-fibroblasts ( = 0.020); and for NF-B-p65 at 4 and 24 h in 16HBE ( = 0.015 and = 0.0002). In the peripheral lung tissue of a representative COPD patient, alveolar type II epithelial cells (TTF-1+) coexpressing c-Kit (CD117) and PCNA were occasionally observed. These data show an increase of cell proliferation induced by a low dosage of extracorporeal shock waves in 16HBE cells and primary bronchial fibroblasts of COPD and control smoking subjects.
慢性阻塞性肺疾病（COPD）是由于小气道结构改变和狭窄以及实质破坏（由于肺气肿导致肺泡附着丧失），所有这些都会导致气流受限。体外冲击波（ESW）可增加细胞增殖和结缔组织成纤维细胞的分化。迄今为止，尚无关于 ESW 治疗 COPD 和对照受试者人支气管成纤维细胞和上皮细胞的研究。我们从 3 名轻度/中度 COPD 患者和 3 名肺功能正常的对照吸烟者的支气管活检中获得了原代支气管成纤维细胞。还研究了 16HBE 细胞。用压电冲击波发生器在低能量（0.3 mJ/mm，500 脉冲）下处理细胞。治疗后，评估细胞活力，并重新培养细胞，在 4、24、48 和 72 小时后进行跟踪。通过 WST-1 试验评估细胞生长，通过 qRT-PCR 在细胞裂解物中以及通过 ELISA 试验在细胞上清液和细胞裂解物中分析增殖标志物。在 ESW 治疗后，我们观察到所有细胞类型的细胞增殖均显著增加。在 16HBE 细胞中，C-Kit（CD117）mRNA 在 4 小时时显著增加。在 16HBE 中，c-Kit（CD117）的蛋白水平在 4 小时时显著增加（<0.0001），在 COPD-成纤维细胞中在 24 小时时显著增加（=0.037）；在 16HBE 中，PCNA 在 4 小时时显著增加（=0.046）；在 CS-成纤维细胞中，Thy1（CD90）在 24 小时和 72 小时时分别显著增加（=0.031 和=0.041）；在 CS-成纤维细胞中，TGF1 在 72 小时时显著增加（=0.038）；在 COPD-成纤维细胞中，原胶原蛋白-1 在 4 小时时显著增加（=0.020）；在 16HBE 中，NF-B-p65 在 4 小时和 24 小时时分别显著增加（=0.015 和=0.0002）。在代表性 COPD 患者的肺外周组织中，偶尔观察到肺泡 II 型上皮细胞（TTF-1+）共表达 c-Kit（CD117）和 PCNA。这些数据显示，低剂量的体外冲击波可诱导 16HBE 细胞和 COPD 和对照吸烟受试者的原代支气管成纤维细胞的细胞增殖增加。
