清热、温补不同治法干预肿瘤相关炎症的时机选择及其调控TAMsNK-kB信号通路的机制研究

Tumor associated inflammation is an important factor to promote tumorigenesis, development,and performs pathological characteristics of dynamic changes in different stages of tumor. whether noxious heat or whole body function decline was more important in the dynamic change of inflammation and how to define the timing of heat-clearing therapy or warming-supplement therapy for tumor control, this problem remains to be elucidated. In order to solve the above problem, inflammation associated classic gastric tumor will be used as the research object for the study,through construction of mouse gastric precancerous lesions and luciferase ( Luc ) transfected human gastric cancer cell SGC-7901 situ xenograft animal model, dynamic changes of mice gastric cancer associated inflammatory will be staged,groups randomized, Dahuang Huanglian Xiexin decoction and Fuzhi Lizhong Decoction respectively for intervention, the change of gastric precancerous lesions and The inhibition rate of tumor is observed ,expression levels of inflammatory cytokines,phenotypic variation of TAMs and NF-kB-P65 expression is detected. The best timing choice will be evaluated for heat-clearing,warming-supplement different therapy on tumor associated inflammation. for studying the mechanism of Dahuang huanglian Xiexin and Fuzhi Lizhong decoction ,Isolation and culture of TAMs,detection of NF-kB signaling pathway activation and expression of the regulation protein by using molecular biology and immune molecular technology. This study will provide scientific basis for the intervention measures of traditional Chinese medicine on tumor associated inflammation.

肿瘤相关炎症是促进肿瘤发生、发展的重要因素，在肿瘤的不同阶段表现为动态变化的病理特征。在炎症的动态变化中热毒或全身功能不足的表现孰轻孰重，如何确定清热和温补不同治法的干预时机更有利于肿瘤的控制，这一问题仍有待阐明。为解决上述问题，本课题以炎症相关性经典肿瘤胃癌为研究对象，通过构建小鼠胃癌前病变、荧光素酶（Luc）转染SGC-7901原位移植瘤动物模型，建立小鼠胃癌相关炎症的动态变化分期，随机分组，予大黄黄连泻心汤和附子理中丸分别进行干预，观察癌前病变的变化和抑瘤率，检测各组组织中炎性因子的表达水平，肿瘤相关巨噬细胞的表型变化及NF-kB相关蛋白的表达水平, 分析清热、温补不同治法对肿瘤相关炎症的最佳干预时机。分离和培养TAMs，通过检测TAMsNF-kB的活性及各调控蛋白的表达情况，明确大黄黄连泻心汤和附子理中丸调控肿瘤相关炎症的机制。这一研究将为肿瘤相关炎症的干预对策提供科学依据。

肿瘤炎症微环境是促进肿瘤增殖和转移的重要因素，在肿瘤的不同阶段表现为动态变化的特征，存在一个虚实转换的过程。清热和温补是中医药抗炎的两大治则，如何确定清热和温补不同治法的干预时机更有利于肿瘤的控制，这一问题仍有待阐明。本研究通过建立胃癌前病变和裸鼠移植瘤模型，运用大黄黄连泻心汤和附子理中丸进行干预，通过观察胃癌前病变的病理学变化及组织中炎症通路相关分子的表达，并通过建立巨噬细胞和肿瘤细胞的transwell共培养体系，模拟体内肿瘤微环境进一步验证，两种不同治法调控巨噬细胞极化及其与NF-κB分子的活性相关性。研究结果表明，大黄黄连泻心汤和附子理中丸均有一定的抗炎作用。在胃癌前病变阶段，大黄泻心汤较附子理中丸在逆转病变方面更为显著，并伴有炎性因子和NF-κB活性的抑制；而肿瘤已经形成阶段，附子理中丸较大黄黄连泻心汤则表现为炎症和免疫的双向调节作用，可能更有利于肿瘤的控制。在体外研究中，大黄黄连泻心汤具有促进巨噬细胞向M2型转化的能力，而附子理中丸与此相反，此效应可能与不同治法对NF-κB活性调节的特点有关。本项目研究不仅进一步验证了肿瘤炎症和免疫的动态发展过程，同时也为肿瘤相关炎症的中医药干预提供了科学依据。