Xu Yu-Dong, Wang Yu, Yin Lei-Miao, Peng Ling-Ling, Park Gyoung-Hee, Yang Yong-Qing
Laboratory of Molecular Biology, Shanghai Research Institute of Acupuncture and Meridian, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, 650 South Wanping Road, Shanghai, 200030, China.
Biol Res. 2017 Jun 21;50(1):23. doi: 10.1186/s40659-017-0128-5.
Airway remodeling is a key feature of asthma, characterized by increased proliferation of airway smooth muscle cells (ASMCs). S100A8 is a calcium-binding protein with a potential to regulate cell proliferation. Here, the effect of exogenous S100A8 protein on the proliferation of ASMCs induced by platelet-derived growth factor (PDGF) and the underlying molecular mechanism was investigated.
Rat ASMCs were cultured with or without a neutralizing antibody to the receptor for advanced glycation end-products (RAGE), a potential receptor for S100A8 protein. Purified recombinant rat S100A8 protein was then added into the cultured cells, and the proliferation of ASMCs induced by PDGF was detected by colorimetric-based WST-8 assay and ampedance-based xCELLigence proliferation assay. The expression levels of RAGE in ASMCs were analyzed using western blotting assay.
Results showed that exogenous S100A8 inhibited the PDGF-induced proliferation of rat ASMCs in a dose-dependent manner with the maximal effect at 1 μg/ml in vitro. Furthermore, when ASMCs was pre-treated with anti-RAGE neutralizing antibody, the inhibitory effect of S100A8 on PDGF-induced proliferation was significantly suppressed. In addition, neither the treatment with S100A8 or PDGF alone nor the pre-treatment with rS100A8 followed by PDGF stimulation affected the expression levels of RAGE.
Our study demonstrated that S100A8 inhibits PDGF-induced ASMCs proliferation in a manner dependent on membrane receptor RAGE.
气道重塑是哮喘的关键特征，其特点是气道平滑肌细胞（ASMCs）增殖增加。S100A8是一种钙结合蛋白，具有调节细胞增殖的潜力。在此，研究了外源性S100A8蛋白对血小板衍生生长因子（PDGF）诱导的ASMCs增殖的影响及其潜在的分子机制。
将大鼠ASMCs与针对晚期糖基化终产物受体（RAGE，一种潜在的S100A8蛋白受体）的中和抗体一起或不一起培养。然后将纯化的重组大鼠S100A8蛋白添加到培养的细胞中，并通过基于比色法的WST-8测定和基于阻抗的xCELLigence增殖测定来检测PDGF诱导的ASMCs增殖。使用蛋白质印迹法分析ASMCs中RAGE的表达水平。
结果表明，外源性S100A8以剂量依赖的方式抑制大鼠ASMCs的PDGF诱导增殖，体外最大作用浓度为1μg/ml。此外，当用抗RAGE中和抗体预处理ASMCs时，S100A8对PDGF诱导增殖的抑制作用被显著抑制。此外，单独用S100A8或PDGF处理，或先用rS100A8预处理后再用PDGF刺激，均不影响RAGE的表达水平。
我们的研究表明，S100A8以依赖膜受体RAGE的方式抑制PDGF诱导的ASMCs增殖。
