基于HMGB1/TLR4/NF-κB途径探讨藿胆鼻渊丸抑制急性鼻窦炎大鼠炎症反应的研究  

Effect of Huodan Biyuan pills in inhibiting inflammatory response in rats with acute sinusitis based on the high-mobility group box 1/Toll-like receptor 4/nuclear factor-kappa B pathway

作　　者：熊静 吴婷 兰超 田道法 蔺婷[2,3] 何迎春[2,3] 周振峰[1] XIONG Jing;WU Ting;LAN Chao;TIAN Daofa;LIN Ting;HE Yingchun;ZHOU Zhenfeng(Yueyang Hospital of Traditional Chinese Medicine,Yueyang 414000,Hunan,China;Key Laboratory of Traditional Chinese Medicine for Prevention and Treatment of Eye,Ear,Nose and Throat Diseases in Hunan Province,Hunan University of Chinese Medicine,Changsha 410208,Hunan,China;Engineering and Technical Research Center of Traditional Chinese Medicine for Prevention and Treatment of Eye,Ear,Nose and Throat Diseases and Protection of Optical Function in Hunan Province,Changsha 410208,Hunan,China)

机构地区：[1]岳阳市中医医院,湖南岳阳414000 [2]湖南中医药大学中医药防治眼耳鼻咽喉疾病湖南省重点实验室,湖南长沙410208 [3]湖南中医药大学湖南省中医药防治眼耳鼻喉疾病与视功能保护工程技术研究中心,湖南长沙410208

出　　处：《湖南中医杂志》2024年第3期146-152,共7页Hunan Journal of Traditional Chinese Medicine

基　　金：湖南省自然科学基金项目(2021JJ70052);湖南省中医药科研计划项目(2021070)。

摘　　要：目的:观察藿胆鼻渊丸对急性鼻窦炎大鼠炎症的影响及其分子机制。方法:将36只SD大鼠按随机数字表法分为正常组,模型组,阳性药物组,藿胆鼻渊丸低、中、高剂量组,每组各6只。采用鼻腔填塞联合鼻腔滴注脂多糖(LPS)法建立急性鼻窦炎大鼠模型。造模完成后,阳性药物组以阿莫西林克拉维酸钾(4∶1)干混悬剂灌胃(98.4375 g/kg),低、中、高剂量组予藿胆鼻渊丸灌胃(1.26 g/kg、2.52 g/kg、5.04 g/kg),正常组和模型组予0.9%氯化钠注射液灌胃,每只SD大鼠灌胃总体积为3 ml/次,2次/d,连续干预21 d。采用苏木精-伊红染色法(HE)观察大鼠鼻窦黏膜炎症变化;酶联免疫吸附法(ELISA)测定大鼠血清中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)含量;免疫组化(IHC)和蛋白质印迹法(WB)检测IL-1β、IL-6和TNF-α蛋白表达情况;WB检测HMGB1、TOLL样受体4重组蛋白(TLR4)、核转录因子-κB p65(NF-κB p65)和磷酸化NF-κB p65(p-NF-κB p65)蛋白表达情况。结果:与正常组比较,模型组鼻窦黏膜组织中炎症细胞浸润明显,阳性药物组和藿胆鼻渊丸不同剂量组炎症细胞聚集减少。与正常组比较,模型组血清中IL-1β、IL-6、TNF-α含量及IL-1β、IL-6、TNF-α蛋白表达均明显升高(P<0.05),鼻窦黏膜组织中的HMGB1、TLR4、NF-κB p65和p-NF-κB p65蛋白表达均明显升高(P<0.05)。与模型组比较,阳性药物组、藿胆鼻渊丸不同剂量组血清中IL-1β、IL-6、TNF-α含量及IL-1β、IL-6、TNF-α蛋白表达均明显降低(P<0.05),鼻窦黏膜组织中的HMGB1、TLR4、NF-κB p65和p-NF-κB p65蛋白表达均下降(P<0.05)。与阳性药物组比较,高剂量组IL-1β、IL-6、TNF-α含量及IL-1β、IL-6、TNF-α蛋白表达均明显降低(P<0.05),HMGB1、TLR4、NF-κB p65和p-NF-κB p65蛋白表达下降(P<0.05)。结论:藿胆鼻渊丸可以有效抑制急性鼻窦炎大鼠炎症反应,其机制可能是通过下调HMGB1分子,抑制TLR4/NF-κB信号Objective:To investigate the effect of Huodan Biyuan pills on inflammation in rats with acute sinusitis and its molecular mechanism.Methods:A total of 36 Sprague-Dawley rats were divided into normal group,model group,positive drug group,and low-,middle-,high-dose Huodan Biyuan pills groups using a random number table,with 6 rats in each group.Nasal cavity occlusion combined with nasal dripping of lipopolysaccharide were performed to establish a rat model of acute sinusitis.After modeling,the rats in the positive drug group were given the dry suspension of amoxicillin and clavulanate potassium(4∶1)at a dose of 98.4375 g/kg by gavage;the rats in the low-,middle-,high-dose Huodan Biyuan pills groups were given Huodan Biyuan pills by gavage at a dose of 1.26 g/kg,2.52 g/kg,and 5.04 g/kg,respectively;the rats in the normal group and the model group were given 0.9%sodium chloride by gavage at a volume of 3 mL each time,twice a day for 21 consecutive days.HE staining was used to observe the inflammatory changes of sinus mucosa;ELISA was used to measure the serum levels of interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α),and immunohistochemistry and Western blotting were used to measure the protein expression levels of IL-1β,IL-6,TNF-α;Western blotting was used to measure the protein expression levels of high-mobility group box 1(HMGB1),Toll-like receptor 4(TLR4),nuclear factor-kappa B p65(NF-κB p65),and phosphorylated NF-κB p65(p-NF-κB p65).Results:Compared with the normal group,the model group had significant inflammatory cell infiltration in sinus mucosa,while the positive drug group and the Huodan Biyuan pills groups had a reduction in inflammatory cell infiltration.Compared with the normal group,the model group had significant increases in the serum levels and protein expression levels of IL-1β,IL-6,and TNF-α(P<0.05),as well as significant increases in the protein expression levels of HMGB1,TLR4,NF-κB p65,and p-NF-κB p65 in sinus mucosa(P<0.05).Compared with the model group

关 键 词：急性鼻窦炎 藿胆鼻渊丸 HMGB1/TLR4/NF-κB 

分 类 号：R285.5[医药卫生—中药学]