Dong Lei, Li Hongli, Li Minjing, Zhang Liting, Qiu Chenhui, Chen Ye
Department of Respiratory and Critical Care Medicine, The Second Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China.
The Second Clinical Medicine College of Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China.
Evid Based Complement Alternat Med. 2022 Nov 22;2022:4427637. doi: 10.1155/2022/4427637. eCollection 2022.
Some domestic scholars revealed the effectiveness of Wenshen Yiqi Keli (WSYQKL) on chronic obstructive pulmonary disease (COPD). However, the exact mechanism of WSYQKL on COPD is fuzzy and needs further research. We adopted UPLC-Q/TOF-MS to analyze the chemical components of WSYQKL. In experiments, human airway smooth muscle cells (hASMCs) were intervened with 2.5% cigarette smoke extract (CSE), medicine serum of WSYQKL, miR-155 mimic, and FoxO3a silencing. Cell viability, proliferation, migration, and the expressions of miR-155, PCNA, Ki67, p21, p27, and FoxO3a were examined by cell counting kit-8, EdU staining, Transwell assay, scarification assay, qRT-PCR, immunol cytochemistry, and western blot, respectively. The association between miR-155 and FoxO3a was assessed by database and luciferase reporter gene analysis. We identified 47 kinds of chemical compositions of WSYQKL in ESI mode and 42 kinds of components of WSYQKL in ESI mode. The medicine serum of WSYQKL strongly alleviated the proliferation and migration of hASMCs induced by CSE in a concentration-dependent manner. The medicine serum of WSYQKL enhanced the levels of p21, p27, and FoxO3a and weakened PCNA and Ki67 levels in hASMCs induced by CSE with the increase of concentration. MiR-155 mimic or FoxO3a silencing notably advanced CSE-treated HASMC viability, proliferation, migration, and the levels of PCNA and Ki67 and downregulated the levels of p21, p27, and FoxO3a in CSE-triggered hASMCs, which was reversed by WSYQKL-containing serum. Our results described that WSYQKL alleviated the proliferation and migration of hASMCs induced by CSE by modulating the miR-155/FoxO3a axis.
一些国内学者揭示了温肾益气颗粒(WSYQKL)对慢性阻塞性肺疾病(COPD)的疗效。然而,WSYQKL治疗COPD的确切机制尚不清楚,需要进一步研究。我们采用超高效液相色谱-四极杆飞行时间质谱联用仪(UPLC-Q/TOF-MS)分析WSYQKL的化学成分。实验中,用人气道平滑肌细胞(hASMCs)分别给予2.5%香烟烟雾提取物(CSE)、WSYQKL含药血清、miR-155模拟物和FoxO3a基因沉默干预。分别采用细胞计数试剂盒-8、EdU染色、Transwell实验、划痕实验、qRT-PCR、免疫细胞化学和蛋白质免疫印迹法检测细胞活力、增殖、迁移以及miR-155、增殖细胞核抗原(PCNA)、Ki67、p21、p27和FoxO3a的表达。通过数据库和荧光素酶报告基因分析评估miR-155与FoxO3a之间的关联。我们在电喷雾离子化(ESI)模式下鉴定出WSYQKL的47种化学成分和42种成分。WSYQKL含药血清以浓度依赖的方式强烈减轻CSE诱导的hASMCs的增殖和迁移。随着浓度增加,WSYQKL含药血清提高了CSE诱导的hASMCs中p21、p27和FoxO3a的水平,并降低了PCNA和Ki67的水平。miR-155模拟物或FoxO3a基因沉默显著提高了CSE处理的人气道平滑肌细胞(HASMC)的活力、增殖、迁移以及PCNA和Ki67的水平,并下调了CSE诱导的hASMCs中p21、p27和FoxO3a的水平,而含WSYQKL的血清可逆转这种情况。我们的结果表明,WSYQKL通过调节miR-155/FoxO3a轴减轻CSE诱导的hASMCs的增殖和迁移。