大黄黄连泻心汤抑制NLRP3炎症小体活性对急性胃溃疡的保护作用及机制研究

Protective effect and mechanism of Dahuang Huanglian Xiexin Decoction on acute gastric ulcer by inhibiting NLRP3 inflammatome activity

[目的]研究不同浸渍条件下大黄黄连泻心汤抑制NOD样受体热蛋白结构域相关蛋白3(NLRP3)炎症小体活性对急性胃溃疡小鼠的保护作用,探讨其作用机制并筛选最佳浸渍条件。[方法]对小鼠进行无水乙醇灌胃以建立急性胃溃疡模型,并给予不同浸提条件下的大黄黄连泻心汤进行实验,同时选用MCC950抑制剂作为阳性药。对各组小鼠进行胃组织苏木精-伊红(HE)染色,观察胃组织病理变化;统计溃疡指数(UI);用酸碱度(pH)试纸测试胃液pH值;检测胃液胃蛋白酶活性(PA);采用酶联免疫吸附(ELISA)法检测血清中白细胞介素1β(IL-1β)和白介素18(IL-18)的水平;采用蛋白免疫印迹法(Western blot)检测药物组小鼠胃组织中NLRP3、半胱氨酸蛋白酶-1(caspase-1)的蛋白表达。[结果]与空白组相比,模型组小鼠胃组织出现明显的溃疡病变,各项指标异常。与模型组相比,药物组可降低乙醇诱导下急性胃溃疡模型小鼠的溃疡指数(P<0.05或P<0.01),抑制胃酸的分泌(P<0.05),抑制胃蛋白酶活性(P<0.01);各药物组中促炎细胞因子IL-1β、IL-18的表达明显降低(P<0.01);通过各药物组灰度值/对照模型组蛋白灰度值的比较,表明药物组能够降低NLRP3、Caspase-1蛋白表达(P<0.01或P<0.05)。最后,对不同浸渍条件下大黄黄连泻心汤的药效进行综合评分,结果表明E(85℃、15 min)组评分最高。[结论]大黄黄连泻心汤对无水乙醇灌胃诱导的急性胃溃疡小鼠有保护作用,其机制可能与抑制NLRP3炎症小体活性有关。通过各药物组抑制NLRP3炎症小体的表达差异,得出85℃、15 min的最佳浸渍条件。

[Objective]Study on the protective effect of Dahuang Huanglian Xiexin Decoction on acute gastric ulcer mice by inhibiting NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome under different immersion conditions and explore its mechanism of action and screen the optimal impregnation conditions.[Methods]Acute gastric ulcer model was established in mice by gavage with anhydrous ethanol,and Dahuang Huanglian Xiexin Decoction was administered under different extraction conditions for the experiment,while MCC950 inhibitor was selected as the positive drug.Hematoxylin-eosin(HE)staining of gastric tissue was performed on each group of mice to observe pathological changes in gastric tissue;Statistical ulcer index(UI);Use a pH test paper to test the potential of hydrogen(pH)value of gastric juice;Detection of gastric juice pepsin activity(PA);Detection of the level of interleukin-1β(IL-1β)and interleukin-18(IL-18)in serum using enzyme-linked immunosorbent assay(ELISA);Western blot was used to detect the protein expression of NLRP3 and Caspase-1 in the gastric tissue of drug treated mice.[Results]Compared with the blank group,the model group mice showed significant ulcerative lesions in the gastric tissue and abnormal indicators.Compared with the model group,the drug group can reduce the ulcer index of ethanol induced acute gastric ulcer model mice(P<0.05 or P<0.01),inhibit gastric acid secretion(P<0.05)and pepsin activity(P<0.01).By comparing the grayscale values of various drug groups with the grayscale values of control model proteins,it was found that the drug group was able to reduce the expression of NLRP3 and Caspase-1 proteins(P<0.05 or P<0.01).Finally,a comprehensive evaluation was conducted on the efficacy of Dahuang Huanglian Xiexin Decoction under different soaking conditions,and the results showed that the E(85℃,15 min)group had the highest score.[Conclusion]Dahuang Huanglian Xiexin Decoction has a protective effect on acute gastric ulcer mice induced by anhydrous ethanol gavage,and its mechanism may be related to the inhibition of NLRP3 inflammasome activity.By inhibiting the expression differences of NLRP3 inflammasomes among different drug groups,the optimal impregnation conditions of 85℃and 15 minutes were obtained.