基于口腔菌群调控的NF-κB通路介导的巨噬细胞极化及炎性因子转录研究复方溃疡凝胶改善放化疗所致口腔黏膜炎的作用机制

放化疗性口腔黏膜炎是影响生活质量，妨碍患者接受放化疗的重要原因，西医尚无有效治疗手段。我们前期临床研究证实复方溃疡凝胶可减轻放化疗性口腔黏膜炎，降低继发性细菌感染发生率；基础研究表明其可降低炎性因子IL-6及TNF-α的水平，提高唾液EGF含量。但因缺乏对启动炎性因子转录的上游NF-kB信号通路活性及激活NF-κB的关键因素-口腔细菌的评估，不能说明其疗效机制。因此，本课题拟从口腔微生态入手，提出复方溃疡凝胶通过调节口腔菌群从而降低NF-κB信号通路活性进而抑制 M1巨噬细胞极化及炎性细胞因子转录来抑制炎症，促进损伤修复的假说。拟建立SPF级、无菌及NF-κB基因敲除小鼠放化疗所致口腔黏膜炎模型，比较中药干预后小鼠口腔黏膜巨噬细胞浸润数量及表型，巨噬细胞极化，炎性因子及生长因子分泌情况，菌群及NF-κB通路相关分子变化，确证假说，同时体外建立上皮细胞与巨噬细胞共培养体系验证该理论。

Chemo- or radio- therapy induced oral mucositis(OM) can decrease the quality of life and even stop patients from chemo- or radio- therapy treatment. No effective conventional treatment has been developed. It is well known that radiotherapy or cytotoxic chemotherapy tends to cause oral microbiome dysbiosis and upregulate NF-kB signaling pathway. Thus, inhibiting oral microbiome induced NF-kB activation and improve epithelium regeneration will mitigate chemo- or radio- induced injury and improve the outcome of both radio- and chemo- therapeutic treatment. Our previous study has revealed that "Compound ulcer gel" can downregulate the pro-inflammatory cytokines and elevate the EGF level in oral mucosa. Here, we hypothesized that "Compound ulcer gel" will improve oral mucositis by inhibiting M1 macrophage polarization and pro-inflammatory cytokine transcription by alteration of oral microbiome and downregulate the activation of NF-kB pathway. In this study, we will test the effect of "Compound ulcer gel" on chemo- and radido- therapy induced OM in mouse model in the context of microbiome change and NF-kB alteration. We will also examine the impact of NF-kB on macrophage phenotype alteration and cytokine transcription. The epithelial cell and macrophage co-culture system will be used to investigate the effect of "Compound ulcer gel" on macrophage polarization and epithelial cell cell reversion mechanism in NF-kB genetic modified macrophage.