香砂六君子汤通过抑制NLRP3炎症小体介导的细胞焦亡预防乙醇诱导的大鼠胃溃疡  

Xiang-Sha-Liujunzi decoction prevents ethanol-induced gastric ulcer in rats by inhibiting NLRP3 inflammasome-mediated pyroptosis

作　　者：张帆 周璇 卓洪民 陈妍 陈永红 宋厚盼 ZHANG Fan;ZHOU Xuan;ZHUO Hongmin;CHEN Yan;Chen Yonghong;SONG Houpan(College of Traditional Chinese Medicine,Hunan Provincial Key Laboratory of Traditional Chinese Medicine Diagnostics,Institute of Traditional Chinese Medicine Diagnostics,Hunan University of Chinese Medicine,Changsha 410208,China;Medical School,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学中医学院,中医诊断学湖南省重点实验室,中医诊断研究所,湖南长沙410208 [2]湖南中医药大学医学院,湖南长沙410208

出　　处：《中国病理生理杂志》2025年第3期534-544,共11页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.82374429);中华中医药学会青年人才托举工程项目(No.2024-QNRC2-B25);湖南省自然科学基金资助项目(No.2023JJ30460);湖南省卫生健康高层次人才项目(No.20240304116)。

摘　　要：目的:探讨香砂六君子汤(Xiang-Sha-Liujunzi decoction,XSLJZD)是否通过抑制核苷酸结合寡聚化结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)炎症小体激活介导的细胞焦亡预防乙醇诱导的大鼠胃溃疡(gastric ulcer,GU)。方法:将大鼠随机分为5组,即正常对照组、模型组、阳性对照(0.33 mg/kg雷贝拉唑)组、低剂量(3.40 g/kg)XSLJZD组和高剂量(6.80 g/kg)XSLJZD组,每组12只。造模前,各组大鼠以灌胃方式分别给予相应药物,连续给药3 d,除正常对照组外,其他组采用无水乙醇法灌胃制备GU模型。检测大鼠溃疡指数和溃疡抑制率;苏木精-伊红染色观察各组动物胃组织病理学变化;过碘酸-希夫染色检测胃组织黏液含量;扫描电镜和透射电镜观察大鼠胃组织超微结构;酶联免疫吸附法检测白细胞介素4(interleukin-4,IL-4)、IL-10、IL-1β、IL-18、前列腺素E2(prostaglandin E2,PGE2)和一氧化氮(nitric oxide,NO)含量;可见光分光度法检测丙二醛(malondialdehyde,MDA)、谷胱甘肽(glutathione,GSH)和超氧化物歧化酶(superoxide dismutase,SOD)水平;免疫荧光染色检测NLRP3、含caspase募集结构域的凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a caspase recruitment domain,ASC)、caspase-1、消皮素D(gasdermin D,GSDMD)、磷酸化核因子κB(phosphorylated nuclear factor-κB,p-NF-κB)和磷酸化NF-κB抑制因子β(phosphorylated NF-κB inhibitorβ,p-IκBβ)蛋白水平。结果:与模型组相比,XSLJZD能明显减轻GU大鼠胃组织损伤,降低溃疡指数,提升溃疡抑制率,显著提高胃组织SOD、GSH、IL-4、IL-10、PGE2和NO水平,显著减少MDA、IL-1β和IL-18含量,显著降低胃组织NLRP3、ASC、caspase-1、GSDMD、p-NF-κB和p-IκBβ蛋白水平。结论:XSLJZD对乙醇诱导的大鼠GU具有较好的预防作用,其机制可能与抑制NLRP3炎症小体激活,减轻炎症反应,遏制胃黏膜上皮细胞焦亡,提高机体抗氧化水平�AIM:This study aimed to investigate whether Xiang-Sha-Liujunzi decoction(XSLJZD)prevents ethanol-induced gastric ulcer(GU)in rats by inhibiting nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome activation-mediated pyroptosis.METHODS:A total of 60 rats were randomly divided into 5 groups:normal control group,model group,positive control(0.33 mg/kg rabeprazole)group,low-dose(3.40 g/kg)XSLJZD group and high-dose(6.80 g/kg)XSLJZD group,with 12 rats per group.Prior to the induction of GU,all rats received their respective treatments via intragastric gavage for 3 d.The GU model was established in all groups except the normal control group using the anhydrous ethanol method.The ulcer index and ulcer inhibition rate were measured.Hematoxylin-eosin staining was performed to observe pathological changes of gastric tissues,and periodic acid-Schiff staining was used to evaluate the mucus content.Scanning electron microscopy and transmission electron microscopy were employed to inves-tigate ultrastructural changes of gastric tissues.Enzyme-linked immunosorbent assay was conducted to measure the levels of interleukin-4(IL-4),IL-10,IL-1β,IL-18,prostaglandin E2(PGE2)and nitric oxide(NO).Additionally,visible spectrophotometric assay was used to determine the levels of malondialdehyde(MDA),glutathione(GSH)and superoxide dismutase(SOD).The protein levels of NLRP3,apoptosis-associated speck-like protein containing a caspase recruitment domain(ASC),caspase-1,gasdermin D(GSDMD),phosphorylated nuclear factor-κB(p-NF-κB)and phosphorylated NF-κB inhibitorβ(p-IκBβ)were assessed using immunofluorescence staining.RESULTS:Compared with model group,XSLJZD significantly alleviated gastric tissue injury,reduced the ulcer index,and enhanced the ulcer inhibition rate in GU rats.Treatment with XSLJZD markedly increased the levels of SOD,GSH,IL-4,IL-10,PGE2 and NO in gas-tric tissue,but significantly decreased the levels of MDA,IL-1βand IL-18.At the molecular level,XSLJZD significantly down-regulated the p

关 键 词：香砂六君子汤 胃溃疡 NLRP3炎症小体 炎症 细胞焦亡 

分 类 号：R285.5[医药卫生—中药学] R573.1[医药卫生—中医学] R363