Department of Pediatrics, Shanghai Baoshan Hospital of Integrated Traditional Chinese and Western Medicine, 201999 Shanghai, China.
Department of Medical Imaging, Shanghai Seventh People's Hospital, 200137 Shanghai, China.
Discov Med. 2024 Feb;36(181):323-331. doi: 10.24976/Discov.Med.202436181.30.
Childhood asthma is a chronic inflammatory disease of the respiratory tract characterized by bronchial inflammation, airway hyperresponsiveness, airflow disorder, and obstruction. Secreted frizzled-related protein 5 (SFRP5) may be associated with respiratory inflammatory diseases. This study investigated the effect of SFRP5 on human airway smooth muscle cells (HASMCs) to provide new ideas for treating asthma.
A total of 30 children with asthma and 30 children who had a physical examination at the same time were selected and divided into asthma and healthy groups. Serum SFRP5 levels were determined by enzyme-linked immunosorbent assay (ELISA) and real-time quantitative polymerase chain reaction (RT-qPCR). Lipofectamine 2000™ regent was used to transfect the overexpression plasmid (pc-) or corresponding negative control (pc-NC) into HASMCs. HASMCs were treated with 10 μg/L platelet-derived growth factor-BB (PDGF-BB), which is an inducer to mimic the asthma-like condition at the cellular level of childhood asthma. HASMCs were divided into control, PDGF-BB (PDGF-BB treatment), PDGF-BB+pc-NC (pc-NC transfection and PDGF-BB treatment), and PDGF-BB+pc- (pc- transfection and PDGF-BB treatment) groups. Cell proliferation was measured by 5-ethynyl-2'-deoxyuridine (EdU) and cell counting kit-8 (CCK-8) assay. Cell migration was detected by Transwell assay. The protein expression was detected by western blot.
Serum SFRP5 expression in the asthmatic group was decreased versus the healthy group ( < 0.0001). Induction of PDGF-BB decreased SFRP5 expression in HASMCs ( < 0.01). SFRP5 expression in the pc- group was increased ( < 0.01). The proliferation and migration of HASMCs increased after PDGF-BB treatment ( < 0.001, < 0.0001), indicating that the asthma model was successfully inducted . Moreover, the expression of β-catenin, cellular-myelocytomatosis viral oncogene (c-Myc), and cyclinD1 proteins in HASMCs increased after PDGF-BB treatment ( < 0.0001). overexpression partly inhibited PDGF-BB-induced proliferation, migration, and expressions of β-catenin, c-Myc, and cyclinD proteins in HASMCs ( < 0.01, < 0.001, < 0.0001).
Serum SFRP5 expression decreases in children with asthma. overexpression partially inhibits PDGF-BB-induced HASMC proliferation and migration by regulating the wingless-type mouse mammary tumor virus (MMTV) integration site family (Wnt)/β-catenin pathway.
儿童哮喘是一种以支气管炎症、气道高反应性、气流障碍和阻塞为特征的呼吸道慢性炎症性疾病。卷曲相关蛋白 5（SFRP5）可能与呼吸道炎症性疾病有关。本研究探讨了 SFRP5 对人气道平滑肌细胞（HASMC）的影响，为哮喘的治疗提供了新思路。
选择同期就诊的 30 例哮喘患儿和 30 例体检儿童，分为哮喘组和健康组。采用酶联免疫吸附试验（ELISA）和实时定量聚合酶链反应（RT-qPCR）检测血清 SFRP5 水平。用 Lipofectamine 2000TM 转染试剂将过表达质粒（pc-）或相应的阴性对照（pc-NC）转染入 HASMCs。用 10μg/L 血小板衍生生长因子-BB（PDGF-BB）处理 HASMCs，以模拟儿童哮喘细胞水平的哮喘样状态。HASMCs 分为对照组、PDGF-BB（PDGF-BB 处理组）、PDGF-BB+pc-NC（pc-NC 转染和 PDGF-BB 处理组）和 PDGF-BB+pc-（pc-转染和 PDGF-BB 处理组）。通过 5-乙炔基-2'-脱氧尿苷（EdU）和细胞计数试剂盒-8（CCK-8）检测细胞增殖。通过 Transwell 检测细胞迁移。通过 Western blot 检测蛋白表达。
哮喘组血清 SFRP5 表达较健康组降低（<0.0001）。PDGF-BB 诱导降低 HASMCs 中 SFRP5 的表达（<0.01）。pc-组 SFRP5 表达增加（<0.01）。PDGF-BB 处理后 HASMCs 的增殖和迁移增加（<0.001，<0.0001），表明哮喘模型成功诱导。此外，PDGF-BB 处理后 HASMCs 中β-catenin、细胞髓样白血病病毒癌基因（c-Myc）和细胞周期蛋白 D1 蛋白的表达增加（<0.0001）。过表达部分抑制 PDGF-BB 诱导的 HASMC 增殖、迁移和β-catenin、c-Myc 和细胞周期蛋白 D 蛋白的表达（<0.01，<0.001，<0.0001）。
儿童哮喘患者血清 SFRP5 表达降低。过表达通过调节 Wnt/β-catenin 通路部分抑制 PDGF-BB 诱导的 HASMC 增殖和迁移。
