文献类型：期刊文献

中文题名：基于NLRP3/ASC/Caspase-1途径探讨清热散浊饮对痛风性关节炎合并高尿酸血症模型大鼠的作用机制

英文题名：Qingre Sanzhuo Decoction Treats Gouty Arthritis Combined with Hyperuricaemia in Rats via NLRP3/ASC/Caspase-1 Pathway

作者：李浩林[1];柏倩[1];程伟刚[1];李伟青[2];杨娟娟[1];贺佩鑫[1];杨会军[2];王海东[2]

第一作者：李浩林

机构：[1]甘肃中医药大学,兰州730000;[2]甘肃省中医院,兰州730050

第一机构：甘肃中医药大学

年份：2025

卷号：31

期号：3

起止页码：49

中文期刊名：中国实验方剂学杂志

外文期刊名：Chinese Journal of Experimental Traditional Medical Formulae

收录：;北大核心:【北大核心2023】；

基金：国家中医药管理局高水平重点学科建设项目(国中医药人教函[2022]226号);甘肃省中医药项目(GZKP-2022-17);王海东甘肃省名中医传承工作室建设项目(甘卫中医函[2022]50号)。

语种：中文

中文关键词：清热散浊饮;NOD样受体蛋白3(NLRP3)/凋亡相关斑点样蛋白(ASC)/胱天蛋白酶-1(Caspase-1)通路;痛风性关节炎;高尿酸血症

外文关键词：Qingre Sanzhuo decoction;NOD-like receptor protein 3(NLRP3)/apoptosis-associated speck-like protein containing CARD(ASC)/cysteinyl aspartate-specific protease 1(Caspase-1)pathway;gouty arthritis;hyperuricaemia

摘要：目的:探讨清热散浊饮治疗痛风性关节炎(GA)合并高尿酸血症(HUA)的效果及相关作用机制。方法:将60只雄性SD大鼠随机分为正常组、模型组、秋水仙碱组(0.5 mg·kg^(-1))、清热散浊饮低、中、高剂量组(17、34、68 g·kg^(-1)),每组10只,除正常组外,其余大鼠均使用改良后GA合并HUA的造模方法;同时给药组每日下午灌胃相应药物,正常组和模型组灌胃等体积无菌生理盐水。末次给药2 h后测定大鼠血清尿酸(SUA)浓度,计算大鼠造模后0.5、12、24、48 h踝关节肿胀程度并对关节炎症评分,苏木素-伊红(HE)染色观察大鼠踝关节病理组织形态,酶联免疫吸附测定法(ELISA)检测大鼠血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、C反应蛋白(CRP)、白细胞介素-18(IL-18)含量,实时荧光定量聚合酶链式反应(Real-time PCR)检测各组大鼠踝关节滑膜组织中NOD样受体蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)、胱天蛋白酶-1(Caspase-1)、焦孔素D(GSDMD)、核转录因子-κB(NF-κB)mRNA表达水平,蛋白免疫印迹法(Western blot)检测大鼠滑膜组织中NLRP3、ASC、Caspase-1蛋白表达,免疫组化检测关节滑膜组织中GSDMD、NF-κB蛋白表达。结果:与正常组比较,模型组大鼠SUA浓度明显升高(P<0.05),踝关节肿胀程度及关节炎症评分升高(P<0.05),滑膜内血管数量增多,滑膜囊内见炎细胞灶,血清TNF-α、IL-1β、CRP、IL-18含量明显升高(P<0.05),踝关节滑膜组织中NLRP3、ASC、Caspase-1、GSDMD、NF-κB mRNA及蛋白表达水平明显升高(P<0.05);与模型组比较,清热散浊饮中、高剂量组大鼠SUA浓度明显降低(P<0.05),踝关节肿胀程度减轻(P<0.05),关节炎症评分降低(P<0.05),血清TNF-α、IL-1β、CRP、IL-18含量降低(P<0.05),踝关节滑膜组织中NLRP3、ASC、Caspase-1、GSDMD、NF-κB mRNA水平及蛋白表达水平降低(P<0.05),但在改善踝关节病理组织形态方面,仅有高剂量组大鼠踝关节滑膜形态良好,关节软骨未见明显病变。结论:清热散浊饮可能通过下调NLRP3/ASC/Caspase-1通路活性,抑制TNF-α、IL-1β、CRP、IL-18等炎症因子表达,从而发挥防治GA合并HUA的作用。
Objective:To investigate the effect and mechanism of Qingre Sanzhuo decoction in treating gouty arthritis(GA)combined with hyperuricaemia(HUA).Methods:Sixty male SD rats were randomized into normal,model,colchicine(0.5 mg·kg^(-1)),and low-,medium-,and high-dose(17,34,68 g·kg^(-1),respectively)Qingre Sanzhuo decoction groups(n=10).The rats in other groups except the normal group were treated with the modified method for the modeling of GA combined with HUA.The drug intervention groups were administrated with corresponding drugs by gavage in the afternoon every day and the normal group and the model group were administrated with an equal volume of sterile normal saline by gavage.The level of uric acid(SUA)in the serum was measured 2 h after the last administration.The degree of ankle joint swelling was calculated 0.5,12,24,48 h after modeling,and joint inflammation was scored.The pathological changes of ankle joints were observed by hematoxylin-eosin staining,and the serum levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),C reactive protein(CRP),and interleukin-18(IL-18)were measured by enzyme-linked immunosorbent assay.Real-time PCR was performed to determine the mRNA levels of NOD-like receptor protein 3(NLRP3),apoptosis-associated speck-like protein containing CARD(ASC),cysteinyl aspartate-specific protease-1(Caspase-1),gasdermin D(GSDMD),and nuclear factor-kappa B(NF-κB)in the synovial tissue of ankle joints.Western blot was employed to determine the protein levels of NLRP3,ASC,and Caspase-1 in ankle joints.The immunohistochemical method was used to detect the expression of GSDMD and NF-κB in the synovial tissue of ankle joints.Results:Compared with the normal group,the model group showed increased SUA in the serum(P<0.05),ankle joint swelling and joint inflammation(P<0.05),increased number of blood vessels in the synovium,inflammatory cell foci in the synovial bursa,elevated serum levels of TNF-α,IL-1β,CRP,and IL-18(P<0.05),and up-regulated mRNA and protein levels of NLRP3,ASC,Caspase-1,GSDMD,and NF-κB in the synovial tissue of ankle joints(P<0.05).Compared with the model group,the medium-and high-dose Qingre Sanzhuo decoction groups showed reduced SUA in the serum(P<0.05),alleviated ankle joint swelling and joint inflammation(P<0.05),lowered serum levels of TNF-α,IL-1β,CRP,and IL-18(P<0.05),and downregulated mRNA and protein levels of NLRP3,ASC,Caspase-1,GSDMD,and NF-κB in the synovial tissue of ankle joints(P<0.05).However,in terms of ameliorating the pathological changes of ankle joints,only the high-dose Qingre Sanzhuo decoction group showed normal morphology of the synovial membrane of ankle joints and no obvious lesion in the articular cartilage.Conclusion:Qingre Sanzhuo decoction may play a role in preventing and controlling GA combined with HUA by down-regulating the activity of NLRP3/ASC/Caspase-1 pathway and inhibiting the expression of inflammatory cytokines,such as TNF-α,IL-1β,CRP,and IL-18.