摘要
目的:探究黄连解毒汤对变应性鼻炎大鼠黏膜杯状细胞及Toll样受体4(TLR4)/核因子κB(NF-κB)信号通路的影响。方法:卵白蛋白制备变应性鼻炎大鼠模型,模型制作成功后分为:模型组,黄连解毒汤低、中、高剂量组,以及阳性对照组。黄连解毒汤低、中、高剂量组分别按5、10和20 g/kg生药量灌胃,阳性对照组给予丙酸氟替卡松鼻喷雾剂(每侧50μg),对照组和模型组灌胃和喷雾等体积生理盐水,每天1次,连续10 d。造模后及治疗后行为学观察评分,治疗后收集并称量鼻腔分泌物重量;PAS染色观察黏膜杯状细胞情况;HE染色观察鼻黏膜组织形态学变化;ELISA检测鼻黏膜组织中白细胞介素(IL)-4、IL-5水平;RT-qPCR检测黏膜组织中鼠钙激活氯通道3(mCLCA3)和黏蛋白5AC(MUC5AC)的mRNA水平;Western blot实验检测黏膜组织中TLR4和NF-κB p65蛋白的水平。结果:造模后,与对照组相比,模型组,黄连解毒汤低、中、高剂量组,以及阳性对照组行为学评分升高(P<0.05)。模型组鼻黏膜组织嗜酸性粒细胞、中性粒细胞和淋巴细胞浸润严重,炎症浸润明显,出现明显的小血管扩张和间质水肿现象;随着黄连解毒汤剂量的升高,淋巴浸润明显减少,但嗜酸性粒细胞和中性粒细胞浸润仍然存在,炎症浸润缓解。与对照组相比,模型组行为学评分、鼻腔分泌物量、黏膜组织中杯状细胞相对比例、黏膜组织中IL-4和IL-5水平、mCLCA3和MUC5AC的mRNA水平及TLR4和NF-κB p65的蛋白水平显著升高(P<0.05);与模型组相比,黄连解毒汤低、中、高剂量组及阳性对照组行为学评分、鼻腔分泌物量、黏膜组织中杯状细胞相对比例、黏膜组织中IL-4和IL-5水平、mCLCA3和MUC5AC的mRNA水平及TLR4的蛋白水平显著降低(P<0.05),黄连解毒汤中、高剂量组及阳性对照组黏膜组织中NF-κB p65的蛋白水平显著降低(P<0.05)。结论:黄连解毒汤降低变应性鼻炎大鼠鼻黏膜组织杯状细胞的相对比例,可能是通过抑制TLR4/NF-κB信号通路实现的。
AIM:To investigate the effect of Huanglian-jiedu decoction(HLJD)on goblet cells and Toll-like receptor 4(TLR4)/nuclear factor(NF)-κB signaling pathway in allergic rhinitis rats.METHODS:The rat model of allergic rhinitis was made by ovalbumin.The model rats were divided into model group,low-,medium-and high-dose HLJD groups,and positive control drug group.The rats in low-,medium-and high-dose HLJD groups were given different doses(5,10 and 20 g/kg,respectively)of crude drug by intragastric administration,the rats in positive control group was given fluticasone propionate nasal spray(50μg per side),and the rats in control group and model group were given normal saline,once per day for 10 days.The behaviors were observed and scored after modeling and treatment,the weight of nasal secretion was measured after the treatment.The goblet cells in nasal mucosa were observed by periodic acid-Schiff(PAS)staining.The morphological changes of nasal mucosa were observed by hematoxylin-eosin(HE)staining.The interleukin(IL)-4 and IL-5 levels in nasal mucosa were measured by ELISA.The mRNA levels of mouse calcium-activated chloride channel 3(mCLCA3)and mucin 5AC(MUC5AC)were detected by RT-qPCR.The protein expression of TLR4 and NF-κB p65 was determined by Western blot.RESULTS:After modeling,compared with control group,the behavioral scores in model group,low-,medium-and high-dose HLJD groups and positive control group were increased(P<0.05).The eosinophils,neutrophils and lymphocytes in nasal mucosa were seriously infiltrated,inflammatory infiltration was obvious,and obvious small vessel dilatation and interstitial edema in model group were observed.With the increase in the dosage of HLJD,the lymphatic infiltration was obviously relieved,but the eosinophil and neutrophil infiltration still existed,and the inflammatory infiltration was relieved.Compared with control group,the behavioral score,nasal secretion,the relative proportion of goblet cells in the mucosa,the IL-4 and IL-5 levels,the mRNA levels of mCLCA3 and MUC5AC,and the protein expression of TLR4 and NF-κB p65 in the mucosa of model group were increased(P<0.05).Compared with model group,the behavioral score,nasal secretion,the relative proportion of goblet cells in the mucosa,the IL-4 and IL-5 levels,the mRNA levels of mCLCA3 and MUC5AC,and the protein expression of TLR4 in the mucosa of low-,mediumand high-dose HLJD groups and positive control group were increased(P<0.05),and the protein expression of NF-κB p65 in the mucosa of medium-and high-dose HLJD groups and positive control group was decreased(P<0.05).CON⁃CLUSION:Huanglian-jiedu decoction reduces the relative proportion of goblet cells in the nasal mucosa of rats with allergic rhinitis,which may be achieved by inhibiting TLR4/NF-κB signaling pathway.
作者
刘书芹
李桂华
赵娜
刘新明
张洁瑛
王金贵
LIU Shu-qin;LI Gui-hua;ZHAO Na;LIU Xin-ming;ZHANG Jie-ying;WANG Jin-gui(Department of Massage,The First Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300193,China)
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2021年第1期140-145,共6页
Chinese Journal of Pathophysiology
基金
天津中医药大学第一附属医院“拓新工程”基金科研课题项目。
