基于LPS介导的TLR4/MyD88/NF-κB通路探讨协定Ⅰ号方治疗鼻咽癌的机制

项目名称： 基于LPS介导的TLR4/MyD88/NF-κB通路探讨协定Ⅰ号方治疗鼻咽癌的机制

项目编号： No.81503622

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 李燕巍

作者单位： 天津医科大学

项目金额： 18万元

中文摘要： 前期临床及基础研究均显示：协定Ⅰ号方临床论治鼻咽癌疗效较好，且对炎症信号通路上的关键分子有明确抑制作用。故推断清热解毒法可通过抗炎而发挥抗鼻咽癌作用，其机制为抑制LPS介导的TLR4/NF-κB炎症通路。本课题拟从TLR4/MyD88/NF-κB通路探讨该方的作用途径及分子机制。1、抗肿瘤效应研究：建立鼻咽癌动物模型制备药物血清，并培养细胞株，研究组方对体内、体外肿瘤生长、增殖及凋亡的影响。2、探讨机制：应用Real-time PCR技术结合免疫组化、免疫印迹检测药物血清在体内、体外对该通路及相关因子转录活性的影响，明确组方作用途径。3、最后，应用蛋白指纹图谱检测药物血清对鼻咽癌5-8F细胞蛋白质表达谱的影响。本研究不仅为中药组方治疗鼻咽癌提供理论依据，清热解毒法治疗鼻咽癌提供实验证据, 也验证了中医药通过延缓炎症因子浸润，从而抑制“炎癌链”转化，最终发挥抗肿瘤作用的新思路。

中文关键词： 鼻咽癌；协定Ⅰ号方；内毒素；脂多糖受体4；慢性炎症

英文摘要： Literature review and preliminary work shows that No.ⅠFormula attenuates lipopolysaccharide（LPS）-induced the development of nasopharyngeal carcinoma (NPC) responses by suppressing Toll-like receptor 4-nuclear factor kappa B(TLR4/NF-kB)signaling pathways. We investigate the mechanism of antitumor activity by the herbs. We establish NPC animal model to investigate the herbs sera pharmacological effect of NPC on the proliferation, cell cycle and apoptosis in NPC rat model and different NPC cell lines, by using MTT assays and flow cytometry. The expression of mRNA and protein were detected by realtime-PCR and western blot. The activity of NF-κB binding DNA was examined by dual luciferase assay. Furthermore, we investigate the possible effects and changes of mammalian target of TLR4 and its downstream signaling pathway in cells induced by herbs sera. The protein expression profiling in 5-8F cells was used by differential in-gel electrophoresis (DIGE) and MALDI-TOF-MS. The underlying mechanisms may be partly due to suppression of TLR4/NF-κB and its downstream signaling and associated protein. This study will excavate the classical theory of traditional Chinese medicine(TCM) and broadening the treatment system of modern medicine, also improve the innovation level of new high effective drugs on the basis of classical prescriptions in TCM. Integrative medicine will do obtain greater achievements in creating new medicine and pharmacology and make more tremendous contributions for the rejuvenation of TCM and NPC therapy.

英文关键词： Nasopharyngeal Carcinoma;No.ⅠFormula ;Lipopolysaccharide;Toll-like receptor 4;Chronic Inflammation