摘要
目的观察不同浓度的血清淀粉样P物质(SAP)对炎症免疫及氧化应激的影响,并探讨其可能的机制。方法将RAW264.7细胞分成对照组、SAP(1.25 mg/L、2.5 mg/L、5 mg/L)组,不同浓度刺激24 h后,流式细胞术测细胞间黏附分子1(ICAM-1)、Fcγ受体表达量,RT-PCR或ELISA检测炎症因子白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、一氧化氮(NO)及ICAM-1的表达。Western blot检测炎症相关脾酪氨酸蛋白激酶(Syk)、细胞外信号调节激酶(ERK1/2)、磷酸化细胞外信号调节激酶(p ERK1/2)的表达。结果 SAP与RAW264.7细胞共同孵育24 h后,SAP作用无论在基因表达水平还是在蛋白表达水平,均可促进炎症因子IL-1β、TNF-α、ICAM-1和活性氮NO的分泌(P<0.05),且随浓度的增加而增加;机制研究发现SAP对细胞Fcγ受体、p ERK1/2的表达无明显影响,但SAP可剂量依赖增加Fcγ受体下游通路蛋白Syk的表达量(P<0.001)。结论 SAP与Fcγ受体结合后,可增加Syk的表达,增加炎症免疫和氧化应激反应。
Aim To investigate the effect of different concentration of serum amyloid P component（ SAP） on inflammation and oxidative stress and its probable mechanism. Methods RAW264. 7 cells were divided into control and SAP group（ 1. 25 mg / L,2. 5 mg / L,5 mg / L） with different concentrations of SAP co-culturing for 24 hours. Expression of intercellular adhesion molecule 1（ ICAM-1） and Fc gamma receptor（ FcγR） were measured by flow cytometry. ELISA and RT-PCR were used to measure inflammatory factors,IL-1β and TNF-α,ICAM-1 and reactive nitric species,nitric oxide（ NO）. Expression of protein（ Spleen tyrosine kinase,Syk,extracellular signal-regulated kinase,ERK1 / 2 and p ERK1 / 2） related to information was detected by Western blot. Results RAW 264. 7 cells were co-cultured with different concentrations of SAP for 24 hours. Secretions of inflammatory factors,IL-1β and TNF-α（ P0. 05）,ICAM-1 and NO were significantly increased in SAP groups. However,expressions of FcγR and p ERK1 / 2 were not affected by the treatment of SAP. For the key protein in downstream pathways of FcγR,Syk was dose-dependent increased and downstream pathways was activated. Conclusion SAP increased expression of Syk,promoted secretion of inflammatory factors and oxidative stress after combined with Fcγ receptors.
出处
《中国动脉硬化杂志》
CAS
北大核心
2016年第5期447-451,共5页
Chinese Journal of Arteriosclerosis
基金
国家自然科学基金(81370380)
广东省自然科学基金(s2013010014739)
关键词
血清淀粉样P物质
巨噬细胞
炎症免疫反应
氧化应激
Serum Amyloid P Component
Macrophage
Inflammatory and Immune Responses
Oxidative Stress
