加味四妙散对急性痛风性关节炎大鼠miR-223-3p与NLRP3/IL-1β信号通路的影响  

Effect of Modified Simiaosan on miR-223-3p and NLRP3/IL-1βSignaling Pathway in Rats with Acute Gouty Arthritis

作　　者：杜明瑞[1,2] 宋哲 李华燕 王鑫 崔炎 DU Mingrui;SONG Zhe;LI Huayan;WANG Xin;CUI Yan(The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China;The First Clinical Medical College,Henan University of Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学第一附属医院,郑州450000 [2]河南中医药大学第一临床医学院,郑州450000

出　　处：《中国实验方剂学杂志》2024年第14期63-70,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南省中医药传承与创新人才工程(仲景工程)中医药学科拔尖人才项目(CZ0325-15);河南省中医药科学研究专项(2018ZY2060);河南省卫生健康委员会基地专项(2021JDZY083)。

摘　　要：目的:通过研究加味四妙散对急性痛风性关节炎(AGA)大鼠模型miR-223-3p与NOD样受体蛋白3(NLRP3)/白细胞介素-1β(IL-1β)信号通路的影响,探讨加味四妙散对AGA的抗炎机制。方法:选取8周龄雄性SD大鼠72只,按随机数字表法分空白组、模型组、秋水仙碱组(0.3 mg·kg^(-1))、加味四妙散高剂量组(31.75 g·kg^(-1))、加味四妙散中剂量组(15.75 g·kg^(-1))、加味四妙散低剂量组(7.875 g·kg^(-1)),每组12只,除空白组外,其余各组采用Coderre法于大鼠右侧踝关节注射单钠尿酸盐(MSU)晶体混悬液,复制急性痛风性关节炎大鼠模型,给药组分别给予相应药液灌胃,模型组和空白组给予等体积的无菌生理盐水溶液灌胃,连续1周。测量大鼠踝关节周径并计算踝关节肿胀度,苏木素-伊红(HE)染色法检测大鼠踝关节滑膜组织病理形态的变化,采用酶联免疫吸附测定法(ELISA)检测各组大鼠血清中IL-1β、肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)水平,蛋白免疫印迹法(Western blot)检测各组大鼠滑膜组织中NLRP3、胱天蛋白酶-1(Caspase-1)、凋亡相关斑点样蛋白(ASC)的蛋白表达,实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠滑膜组织中NLRP3、Caspase-1、ASC mRNA的表达和miR-223-3p的表达。结果:与正常组比较,模型组大鼠踝关节肿胀度显著升高(P<0.01),滑膜组织结构紊乱,滑膜细胞增生明显,有大量炎症细胞浸润,大鼠血清中IL-1β、IL-6、TNF-α水平显著上升(P<0.01),NLRP3、Caspase-1、ASC的蛋白与mRNA表达增加,miR-223-3p的表达下降;与模型组比较,秋水仙碱组、加味四妙散高、中剂量组关节肿胀度明显降低(P<0.05),秋水仙碱组、加味四妙散高、中、低剂量组滑膜细胞增生及炎性细胞浸润较模型组均有不同程度减轻,其中秋水仙碱组和加味四妙散高剂量组改善最为明显,加味四妙散各剂量组和秋水仙碱组大鼠血清中IL-1β、IL-6、TNF-α水平显著下�Objective:To investigate the effect of modified Simiaosan on miR-223-3p and NOD-like receptor thermal protein domain associated protein 3(NLRP3)/interleukin-1β(IL-1β)signaling pathway in rat model with acute gouty arthritis(AGA)and explore the anti-inflammatory mechanism of modified Simiaosan on AGA.Method:A total of 728-week-old male SD rats were selected.They were divided into blank group,model group,colchicine group(0.3 mg·kg^(-1)),high-dose modified Simiaosan group(31.75 g·kg^(-1)),mediumdose modified Simiaosan group(15.75 g·kg^(-1)),and low-dose modified Simiaosan group(7.875 g·kg^(-1))according to random number table method,with 12 rats in each group.Except for the blank group,monosodium urate(MSU)crystal suspension was injected into the right ankle joint of rats by the Coderre method in other groups to replicate the rat model with AGA.The drug administration groups were given the corresponding drug solution by gavage,and the model group and the blank group were given an equal volume of sterile sodium chloride solution by gavage for one week.The circumference of the rats'ankle joint was measured,and the swelling degree of the ankle joint was calculated.Hematoxylin-eosin(HE)staining was used to detect the pathological morphological changes in the synovial tissue of the ankle joint.The levels of IL-1β,tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)in the serum of rats in each group were detected by enzyme-linked immunosorbent assay(ELISA).Western blot was used to detect the protein expressions of NLRP3,Caspase-1,and apoptosis-related spot-like protein(ASC)in synovial tissue of rats in each group,and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR)was used to detect the mRNA expressions of NLRP3,Caspase-1,and ASC and the expression of miR-223-3p in synovial tissue of rats.Result:Compared with that in the normal group,the swelling degree of the ankle joint in the model group was higher(P<0.01),and the synovial tissue structure was disordered.Synovial cells proliferate

关 键 词：加味四妙散 急性痛风性关节炎 炎症信号通路 miR-223-3p 

分 类 号：R2-0[医药卫生—中医学] R33R289R684.3