从TLR4-NFkB-炎症因子信号通路探讨荆花胃康胶丸及其拆方抗小鼠Hp感染性胃炎的作用机制

H.pylori is one of the major causes of gastric cancer. It is significant for the prevention and treatment of gastric cancer to eradicate H.pylori before atrophic gastritis and intestinal metaplasia. A drug which does both way in raising eradication rate and promoting inflammation subsiding will be an ideal choice for the treatment. Our former researches have found that Jinghua Weikang Capsule was equal to quadruple therapy on eradication rate of H.pylori, in vivo and vitro the drug showed bacteriostasis effects against standard and antibiotic resistance H.pylori strains. However, the former researches rarely discussed the effects on H.pylori infected gastritis of this drug. Our research will establish the mouse model of H.pylori infected gastritis, then inoculate Jinghua Weikang Capsule and its ingrediants with and without western medicine. The impacting effects on gastric mucosal inflammation, ultrastructure of gastric epithelial cells, the transcription and protein expression of important elements in TLR4-NFκB-inflammatory cytokines signaling pathway will be observed by pathological，morphological and molecular biological methods respectively, in order to certify the anti-inflammation action and reveal the posible mechanism of this drug. Our study will complement the fundamental researches on exploring the therapeutical effect and mechanism of Jinghua Weikang Capsule, thus provide evidences for the long-term prevention and treatment of gastric cancer by the integration of traditional Chinese and Western medicine.

H.pylori是胃癌发生的重要危险因素之一，在患者尚未发生萎缩性炎或肠化根除H.pylori并消除炎症对胃癌防治具有重要意义。能够提高H.pylori根除率并促进炎症消退药物将是相关治疗的理想选择。我们前期研究发现，荆花胃康胶丸与三联疗法联合对H.pylori根除率与铋剂四联疗效相当；该药在体内、体外对H.pylori标准及耐药株均有较好的抑杀作用。有关该药对H.pylori感染性炎症作用研究则尚未开展。本研究建立H.pylori感染胃炎小鼠模型，用该药及其拆方单药、中西医联合用药干预，从病理学角度观察其消除胃黏膜炎症的作用，在形态学角度观察胃黏膜上皮细胞超微结构变化，并利用分子生物学方法检测其对TLR4-NFκB-炎症因子通路中重要受体及衔接分子在转录及蛋白水平表达的影响，从而明确该药抗炎作用并揭示其中可能的作用机制，为治疗H.pylori感染性胃炎及胃癌远期中西医结合防治提供依据。

幽门螺杆菌（HP）我国感染率56%以上，已然成为公共卫生健康问题。鉴于Hp耐药率不断上升及临床抗生素用药时间持续延长和数量不断增加的背景，本课题选用临床有确切疗效的中药荆花胃康胶丸，对该药及基于TLR4-NFκB-炎症因子信号通路对荆花胃康胶丸抗H. pylori感染性炎症作用的作用机制进行研究，发现荆花胃康胶丸可以降低小鼠外周血中TNF-α和IL-1ß的水平，改善胃黏膜炎症。荆花胃康胶丸通过下调TLR4-NF-κB信号通路激活，抑制炎症。研究发现：一、C57/BL6J小鼠和昆明小鼠RUT实验阳性率分别为10/10和10/10,病理感染率均为100%，WS银染色阳性率分别为9/10和10/10。两组之间定植率没有统计学差异（P>0.05）。二、荆花胃康降低IκB和NF-κB p65蛋白的磷酸化水平，抑制NF-κB p65蛋白核转移，从而减轻炎症反应。三、荆花胃康抑制炎症表达的作用与剂量有关，50mg/kg和100mg/kg是应用于小鼠的有效剂量，中剂量和高剂量组之间没有显著差异（P>0.05），与低剂量组之间呈现显著差异（P<0.05）。以上结果发表SCI论文3篇，已发表和录用的核心期刊论文8篇，拟投送论著4篇。本研究多次参与国内学术交流，其中参与学术会议报告8次。研究期间培养已毕业博士研究生、硕士研究生各一名，在读博士、硕士研究生2名，已开展进一步深入的研究。研究不仅补充了荆花胃康在HP领域的研究空白，更为临床应用提供了高质量的循证医学证据，为进一步探索中药抗Hp药效提供了基础，培养了多名高素质人才，具有重要意义。