摘要
【目的】检测大鼠放射性口腔黏膜炎形成过程中病损局部组织内细胞因子,探讨其动态变化规律及意义。【方法】获取接受单次30GyX线照射后5d、8d、14d的大鼠口腔黏膜组织,采用抗体芯片技术检测19种细胞因子;荧光定量PCR检测8种炎性相关因子的mRNA水平;ELISA定量检测IL-1α、IFN-γ、TIMP-1。【结果】抗体芯片技术检测到,照射后Frac、IL-4、IL-6、IL-10、TNF-α5种细胞因子水平下降;IL-1β、LIX、VEGF、β-NGF4种细胞因子照射前后水平无明显差异;照射后水平上升幅度<2倍的有CINC-2、GM-CGF、Leptin、MCP-1和MIP-3α5种细胞因子,上升幅度≥2倍的有CINC-3、INF-γ、IL-1α和TIMP-14种细胞因子。Real-timePCR检测到抗凋亡相关基因Bcl-2,以及IL-6、TNF-α、VEGF及IL-1β的mRNA表达水平下降;而促凋亡相关基因Bax、中性粒细胞局部募集相关基因CINC和TLR-9表达上升。IL-1α、IFN-γ、TIMP-1的ELISA检测结果与前两种检测结果一致。【结论】大鼠抗体芯片和荧光定量PCR、ELISA检测结果互相佐证了病损组织内的细胞因子在口腔黏膜炎发生、发展、演进中呈现分泌受阻和表达增加的动态变化并发挥调控作用。
【Objective】 This study was designed to explore the dynamic changes of cytokines in the local tissues of radiation- induced oral mucositis in a rat model. 【methods】 The rat oral mucosal tissues were obtained at 5 d, 8 d, and 14 d after irradiation, which were received single X-ray irradiation of 30 Gy locally. RayBio Rat Cytokine Array was applied to analyze the 19 cytokines. The mRNA levels of the 8 inflammatory-related cytokines were analyzed by real time-PCR. ELISA was employed to detect IL-1α, IFN-γ, and TIMP-1 protein levels. 【Results】 Cytokine array detection showed that cytokines such as Frac, IL-4, IL-6, IL-10, and TNF-α were down-regulated after irradiation. IL-1β, LIX, VEGF, and β-NGF were no obviously changed at the same time course. The elevate range less than 2 fold of the CINC-2, GM-CGF, Leptin, MCP-1, and MIP-3α after irradiation were detected. The increasing range more than 2 fold of CINC-3, INF-γ, IL-1α, and TIMP-1 post irradiation were detected. The mRNA levels of anti-apoptosis, such as Bcl-2, and TNF-α, IL-6, VEGF, and IL-1β were low down, while the levels of promoting apoptosis cytokine such as Bax, and neutrophil recruitment associated cytokines, CINC, and TLR-9 were up-regulated after irradiation. The ELISA results of IL-1α, IFN-γ, and TIMP-1 were identical with the results of the two previous analyses. 【Conclusion】 The results of the three methods in the study should present evidences to prove each other. Some cytokines in the local lesion of oral mucositis during the initiate, development, and progression stages were blocked, while others were positively up-regulated to involving the control of the pathogensis of oral mucositis.
出处
《中山大学学报（医学科学版）》
CAS
CSCD
北大核心
2010年第2期231-237,共7页
Journal of Sun Yat-Sen University:Medical Sciences
基金
广东省科技计划项目(2005B50301004
2006B36006010)
关键词
放射性口腔黏膜炎
细胞因子
抗体芯片
radiation-induced oral mucositis
cytokines
antibody array
