基于巨噬细胞M1/M2分布格局动态变化研究彝药美洲大蠊调控UC癌变发展的机制

Ulcerative colitis (UC) is a severe risk to human health，which has been by WTO. It is a precancerous disease and a hot spot of research in recent years. National medicines play a unique role in anti-inflammatory and repair. Preliminary experiments showed that the active fraction possessed satisfactory therapeutic effects towards acute-, chronic-, and immuno-ulcerative colitis rats. in addition, the main components of the active fraction were polypeptides BII, which come form Yi minority medicine Periplaneta americana L..We have proposed a hypothesis that the polypeptides BII may paly a bidirectional immunomodulatory role at different stages. By regulating the distribution pattern of macrophages, it could play an initial role in antibacterial in the early stages , then anti-inflammatory and enteral wound healing with time, and further inhibit the possibility of canceration.This study is intended to dynamically change the distribution and pattern of M1/M2 in macrophages, which has always been the focus of research at different stages of the course of UC, and explores the mechanism of the treatment of UC.The expression and phenotype distribution, polarization trend and related cytokines and signaling pathways in macrophages of AOM/DSS-UC by dynamic observation of BII, clarify the function of macrophages in the regulation of BII dynamic law, so as to determine its effect on UC process and mechanism.

癌前疾病溃疡性结肠炎（UC）一直威胁着人类健康，被WTO列为现代难治病之一，为近年来的研究热点。民族药物在抗炎修复方面有着独特的作用，前期研究发现彝药昆虫美洲大蠊其多肽物质（BII）对大鼠急、慢性及免疫性UC有显著疗效，并推测美洲大蠊多肽物质可能在不同阶段通过调节巨噬细胞分布格局，从而发挥初期抗菌，后期抗炎及修复肠炎创面的双向免疫调节作用，进一步抑制癌变的可能。本研究拟从巨噬细胞M1/M2分布和格局动态变化，这一贯穿于UC病程不同阶段的研究热点，探讨治疗UC的机制研究。通过研究BII调控过AOM/DSS诱导的UC BALB/c小鼠肠道，动态并逐层深入观测巨噬细胞的组织表达情况、表型分布特征、极化趋势、相关细胞因子和信号通路等，阐明BII调控巨噬细胞表达分布及功能变化的动态规律，从而确定其对UC的作用过程及机制。

溃疡性结肠炎（UC）是病因不明的肠道慢性非特异性自身免疫性疾病，其极为难以治愈且癌变风险呈逐年上升的趋势。目前已知巨噬细胞极化在溃疡性结肠炎向结直肠癌进程中具有极为重要的作用。大量的研究报道和临床实践表明，彝药美洲大蠊对消化系炎性疾病和黏膜溃疡有着显著的疗效，然而其对UC的作用机制鲜有报道。因此，本项目通过解析美洲大蠊多肽类物质基础，深入探究其通过动态调节巨噬细胞M1/M2极化格局实现治疗UC向结直肠癌转变（CRC）的机制。本次研究内容及结果如下：（1）采用LC-MS/MS和nano LC-ESI-MS/MS等现代质谱技术和相关数据库检索比对，最终检测到CⅡ-3中小分子化合物1192个，多肽类物质185条；CⅡ-3经过硅胶Sephadex LH20、半制备HPLC等色谱方法分离鉴定得到12个化合物，初步进行了RAW264.7体外活性筛选及诱导极化为M1/M2后对相应炎症因子的mRNA表达水平的影响。（2）美洲大蠊提取物CⅡ-3可显著改善CAC模型小鼠结肠长度、重量、瘤数、组织病理学症状及降低血浆癌胚蛋白表达含量，缓解结肠损伤，动态调节巨噬细胞M1/M2相关炎性细胞因子，抑制肿瘤负荷，从而发挥抗炎及抑制癌变进程的作用；（3）CⅡ-3通过抑制TLR4、MAPK P38的信号传导，动态调节M1/M2平衡，抑制结肠黏膜的过度增生和癌变。本研究可为民族药靶向巨噬细胞极化治疗UC-CRC及临床联合治疗应用提供提供一条新的思路和理论基础，为更好的开发云南特色昆虫药物提供科学依据。