苗药头花蓼通过调控ZAC抑制CLIC4/NLRP3炎症小体活化介导胃上皮保护的分子机制研究

Helicobacter pylori-associated gastritis (HAG) plays a key role in the progressions of gastric cancer, always as the focus of onset studies. Inhibiting the activation of NLRP3 inflammasomes can evidently improve the outcomes of HAG, also depending on CLICs-mediated Cl- outflow. We have previously found that Polygonum capitatum up-regulated the expression of ZAC but down-regulated that of CLIC4, there by blocking the activation of NLRP3 inflammasomes. In combination with up-to-date literatures, we hypothesize that the inhibitory effects of ZAC on CLIC4-mediated activation of NLRP3 inflammasomes play a crucial role in Polygonum capitatum induced gastric epithelial repair. To validate this hypothesis, the contents of this project are as follows. We will prove the correlations between ZAC and CLIC4 by using HAG clinical samples, macrophage models from different sources and mouse models; evaluate the effects of ZAC expression inhibition or overexpression on CLIC4 expression as well as the assembling and activation of NLRP3 inflammasomes; and clarify the regulatory relationships between the two factors by ChIP and luciferase reporter assays. In short, the aim of this project is to provide abundant experimental evidence for the treatment of HAG by elucidating the molecular mechanisms by which Polygonum capitatum repairs the gastric epithelium based on the inhibitory effects of ZAC on CLIC4-mediated activation of NLRP3 inflammasomes.

幽门螺杆菌相关性胃炎（HAG）是胃癌演变过程中的关键环节，一直是胃癌发病研究的重点。研究发现抑制NLRP3炎症小体活化明显改善HAG炎症结局，且依赖于CLICs介导的氯离子外流。课题组前期发现苗药头花蓼上调ZAC表达，降低CLIC4表达，阻滞NLRP3炎症小体活化。结合最新的研究进展，我们提出“ZAC抑制CLIC4介导NLRP3炎症小体活化是头花蓼修复胃上皮细胞分子机制的关键环节”的科学假说。围绕该项目假说，我们将以HAG临床样本、不同来源HAG的巨噬细胞模型和HAG小鼠模型为研究对象，确认ZAC与CLIC4的相关性；通过抑制或过表达ZAC，观察其对CLIC4表达和NLRP3炎症小体组装以及活化的影响；并借助ChIP和荧光素酶报告基因等方法明确两者之间的调控关系。从ZAC抑制CLIC4介导NLRP3炎症小体活化角度阐明头花蓼对胃粘膜损伤修复的分子机制，为HAG的治疗提供丰富的实验依据。

幽门螺杆菌相关性胃炎（HAG）是胃癌演变过程中的关键环节，一直是胃癌发病研究的重点。课题组前期发现苗药头花蓼上调ZAC表达，降低CLIC4表达，阻滞NLRP3炎症小体活化，明显改善HAG炎症结局。结合最新的研究进展，我们提出“ZAC抑制CLIC4介导NLRP3炎症小体活化是头花蓼修复胃上皮细胞分子机制的关键环节”的科学假说。围绕该项目假说，我们将以HAG临床样本、不同来源HAG的巨噬细胞模型和HAG小鼠模型为研究对象，确认ZAC与CLIC4的相关性；通过抑制或过表达ZAC，观察其对CLIC4表达和NLRP3炎症小体组装以及活化的影响；并借助荧光素酶报告基因等方法明确两者之间的调控关系。本课题实验结果表明H.pylori感染胃黏膜上皮细胞或巨噬细胞后，可抑制ZAC表达，CLIC4表达升高并易位到细胞膜上，促使氯离子外流，进一步活化细胞中NLRP3炎症小体，造成胃黏膜的损伤。进一步证实了NLRP3炎症小体的异常活化是HAG的重要启动因素或激活因子，是HAG炎症反应的核心，ZAC和CLIC4是治疗HAG的潜在靶点。此外，我们用头花蓼对HAG的细胞模型和动物模型进行治疗，发现头花蓼能显著改善炎性损伤，促进ZAC的表达上调，降低CLIC4、NLRP3、IL-18和IL-1β表达，抑制NLRP3炎症小体的活化。本课题的研究不仅有助于解释CLICs的功能机制，阐明NLRP3炎症小体激活的相关调控机制，而且从分子机制上证实头花蓼能有效抑制HAG的炎症反应，表明其可作为治疗HAG的候选药物。