加味温胆汤调控NF-κB/NLRP3通路干预炎性反应抗大鼠糖尿病动脉粥样硬化的机制  

Jiawei Wendantang Regulates NF-κB/NLRP3 Pathway to Reduce Inflammation in Rat Model of Diabetic Atherosclerosis

作　　者：刘超[1,2] 王琳 姚凤云[1] 黄艳美 王炳志[1] 崔言坤 LIU Chao;WANG Lin;YAO Fengyu;HUANG Yanmei;WANG Bingzhi;CUI Yankun(Jiangxi University of Chinese Medicine,Nanchang 330004,China;The Affiliated Hospital of Jiangxi University of Chinese Medicine,Nanchang 330006,China)

机构地区：[1]江西中医药大学,南昌330004 [2]江西中医药大学附属医院,南昌330006

出　　处：《中国实验方剂学杂志》2024年第14期71-77,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：江西省教育厅科学技术研究项目(GJJ211253);国家自然科学基金项目(82360897);江西省自然科学基金项目(20212BAB216007);江西省中医药管理局科技计划项目(2022B898)。

摘　　要：目的:通过观察加味温胆汤对糖尿病动脉粥样硬化模型大鼠核转录因子-κB(NF-κB)/NOD样受体蛋白3(NLRP3)通路及其相关炎性因子表达的影响,探讨该方防治糖尿病动脉粥样硬化的机制。方法:将54只SPF级大鼠随机分为空白组、模型组、阿托伐他汀组、加味温胆汤高、中、低剂量组,除空白组外,其余各组均采用腹腔注射链脲佐菌素(STZ)+高糖高脂饲料喂养法建立糖尿病动脉粥样硬化大鼠模型,空白组注射等剂量的柠檬酸缓冲液并喂养普通饲料。加味温胆汤高、中、低剂量组给药剂量分别是18.2、9.1、4.55 g·kg^(-1)·d^(-1),阿托伐他汀组给药剂量为0.9 mg·kg^(-1)·d^(-1),连续给药4周,每隔6 d检测1次大鼠尾静脉血糖,经末次给药后,麻醉取材。采用酶联免疫吸附测定法(ELISA)检测血清白细胞介素-18(IL-18)、C反应蛋白(CRP)、肿瘤坏死因子-α(TNF-α)、细胞间黏附分子-1(ICAM-1)含量。蛋白免疫印迹法(Western blot)检测腹主动脉NF-κB p65、NLRP3、ICAM-1蛋白相对表达水平。实时荧光定量聚合酶链式反应(Real-time PCR)检测腹主动脉NLRP3、白细胞介素-1β(IL-1β)mRNA表达水平。苏木素-伊红(HE)染色法观察胸主动脉病理变化。结果:与空白组比较,模型组大鼠血清IL-18、CRP、TNF-α、ICAM-1及血糖含量明显升高(P<0.05,P<0.01),主动脉NF-κB p65、NLRP3、ICAM-1蛋白表达显著升高(P<0.01),NLRP3、IL-1βmRNA表达明显升高(P<0.05)。与模型组比较,加味温胆汤组能明显降低大鼠血清IL-18、CRP、TNF-α、ICAM-1及血糖的水平(P<0.05,P<0.01),显著降低腹主动脉NF-κB p65、NLRP3、ICAM-1蛋白表达水平(P<0.01),明显降低腹主动脉NLRP3、IL-1βmRNA表达水平(P<0.05,P<0.01),明显改善大鼠胸主动脉病理损伤。结论:加味温胆汤可能通过调控NF-κB/NLRP3信号通路减少炎性因子释放和黏附,调节血糖,发挥抗糖尿病动脉粥样硬化作用。Objective:To explore the mechanism of Jiawei Wendantang in preventing and treating diabetic atherosclerosis by observing the effect of this prescription on the nuclear factor-κB/NOD-like receptor protein 3(NF-κB/NLRP3)pathway and related inflammatory cytokines in rat model of diabetic atherosclerosis.Method:Fifty-four SPF-grade rats were randomized into blank,model,atorvastatin(0.9 mg·kg^(-1)·d^(-1)),and high-,medium-,low-dose(18.2,9.1,4.55 g·kg^(-1)·d^(-1),respectively)Jiawei Wendantang groups.The rats in other groups except the blank group were modeled for diabetic atherosclerosis by intraperitoneal injection of streptozotocin and feeding with a high-sugar high-fat diet,and those in the blank group were injected with an equal dose of citric acid buffer and fed with a regular diet.The drug administration lasted for 4 weeks,and the blood glucose level in the tail vein was measured every 6 days.After the last administration,the rats were anesthetized for sample collection.Enzyme-linked immunosorbent assay was employed to measure the serum levels of interleukin-18(IL-18),C-reactive protein(CRP),tumor necrosis factor-α(TNF-α),and intercellular adhesion molecule-1(ICAM-1).Western blot was employed to determine the relative protein levels of NF-κB p65,NLRP3,and ICAM-1 in the abdominal aorta.Real-time quantitative polymerase chain reaction was employed to determine the mRNA levels of NLRP3 and interleukin-1β(IL-1β)in the abdominal aorta.The pathological changes in the thoracic aorta were observed by hematoxylin-eosin staining.Result:Compared with the blank group,the model group showed elevated levels of IL-18,CRP,TNF-α,and ICAM-1 in the serum and blood glucose(P<0.05,P<0.01),up-regulated protein levels of NF-κB p65,NLRP3,and ICAM-1(P<0.01),and up-regulated mRNA levels of NLRP3 and IL-1β(P<0.05).Compared with model group,Jiawei Wendantang lowered the levels of IL-18,CRP,TNF-α,ICAM-1 and blood glucose(P<0.05,P<0.01),downregulated the protein levels of NF-κB p65,NLRP3,and ICAM-1(P<0.01),and down-regulated

关 键 词：加味温胆汤 糖尿病动脉粥样硬化 核转录因子-κB(NF-κB) NOD样受体蛋白3(NLRP3) 炎症因子 

分 类 号：R2-0[医药卫生—中医学] R22R242R285.5R587.1