摘要
目的:探讨麻黄、细辛、附子配伍对过敏性鼻炎(AR)小鼠肺组织中2型先天淋巴细胞(ILC2s)相关因子表达的影响。方法:54只C57BL/6J小鼠按随机数字表法分为空白组、模型组、麻黄细辛附子汤组、辛附组、麻辛组、麻附组、麻黄组、附子组、细辛组,每组6只。除空白组外,其余组均采用卵清蛋白(OVA)腹腔注射及滴鼻激发制备AR模型。造模成功后,空白组及模型组给予0.2 m L·d^(-1)生理盐水灌胃,麻黄细辛附子汤组(2.31 g·kg^(-1))、辛附组(1.54 g·kg^(-1))、麻辛组(1.16 g·kg^(-1))、麻附组(1.93 g·kg^(-1))、麻黄组(0.77 g·kg^(-1))、附子组(1.16 g·kg^(-1))、细辛组(0.39 g·kg^(-1))小鼠分别灌胃相应药物,连续14 d。观察各组小鼠生存状态并采用酶联免疫吸附测定法(ELISA)检测鼻激发后血清中免疫球蛋白E(Ig E)含量;采用苏木素-伊红(HE)染色法观察小鼠鼻、肺组织的病理学形态变化;采用免疫荧光染色法(IF)检测小鼠肺组织ILC2s表达量;采用实时荧光定量聚合酶链式反应(Real-time PCR)检测小鼠肺组织转录因子GATA结合蛋白3(GATA3)、视黄酸受体相关孤儿受体-α(RORα)、DNA结合抑制因子2(ID2)m RNA表达量;采用ELISA检测血清中Ig E、白细胞介素(IL)-4、IL-5、IL-13水平。结果:与空白组比较,模型组小鼠生存状态差,鼻激发后血清Ig E显著升高(P<0.01);后鼻甲黏膜可见大量中性粒细胞浸润,并伴随明显的鼻黏膜出血及鼻腔脓性分泌物,可见脱落上皮,肺支气管壁显著增厚,血管内充血水肿明显,可见大量炎性细胞散漫浸润,肺泡结构损坏严重,局部肺实变;肺组织ILC2s表达显著升高(P<0.01);GATA3、RORαm RNA表达明显升高,ID2 m RNA表达明显降低(P<0.05,P<0.01);血清Ig E,IL-4、IL-5、IL-13含量明显升高(P<0.05,P<0.01)。与模型组比较,麻黄细辛附子汤及其各拆方组小鼠生存状态改善,鼻、肺组织炎细胞浸润显著减少,少量鼻黏膜出血,气管壁变薄,未见充血、水肿及鼻腔分泌物;肺组织ILC2s表达量显著降低(P<0.01);GATA3 m RNA表达明显降低(P<0.05),其中附子组显著降低(P<0.01),仅有麻附组、麻黄组RORαm RNA表达明显降低(P<0.05);血清Ig E明显降低(P<0.05),IL-5显著降低(P<0.01),除附子组外,其余各组IL-4显著降低(P<0.01),麻黄细辛附子汤组IL-13明显降低(P<0.05),麻附组、麻黄组、附子组、细辛组IL-13显著降低(P<0.01)。结论:麻黄、细辛、附子不同配伍均可减轻OVA诱发的AR小鼠炎症,在降低Ig E、IL-5分泌环节更具优势;麻黄与附子配伍在降低GATA3m RNA、RORαm RNA水平来抑制ILC2s的表达从而发挥抗过敏作用方面最具有优势,而其余配伍在抑制GATA3 m RNA表达环节更具有广泛性优势。
Objective:To explore the effects of compatibility of Ephedrae Herba,Asari Radix et Rhizoma,and Aconiti Lateralis Radix Praeparata on the expression of type 2 innate lymphoid cells(ILC2s)-related factors in the lung of allergic rhinitis(AR)mice.Methods:According to the random number table method,fifty-four C57BL/6J mice were randomly divided into the following groups:Blank group,model group,Mahuang Fuzi Xixintang group,Asari Radix et Rhizoma and Aconiti Lateralis Radix Praeparata group,Ephedrae Herba and Asari Radix et Rhizoma group,Ephedrae Herba and Aconiti Lateralis Radix Praeparata group,Ephedrae Herba group,Aconiti Lateralis Radix Praeparata group,and Asari Radix et Rhizoma group(6 mice in each group).Except the blank group,the other groups were subjected to intraperitoneal injection of ovalbumin(OVA)and intranasal challenge to induce AR.After the AR model was established,the mice in the blank group and the model group were given 0.2 mL·d^(-1)normal saline by gavage,while those in the Mahuang Fuzi Xixintang group(2.31 g·kg^(-1)),Asari Radix et Rhizoma and Aconiti Lateralis Radix Praeparata group(1.54 g·kg^(-1)),Ephedrae Herba and Asari Radix et Rhizoma group(1.16 g·kg^(-1)),Ephedrae Herba and Aconiti Lateralis Radix Praeparata group(1.93 g·kg^(-1)),Ephedrae Herba group(0.77 g·kg^(-1)),Aconiti Lateralis Radix Praeparata group(1.16 g·kg^(-1)),and Asari Radix et Rhizoma group(0.39 g·kg^(-1))were given corresponding medicine by gavage,with the treatment lasting for 14 consecutive days.The survival state of mice in each group was observed,and the levels of serum immunoglobulins E(IgE)after intranasal challenge were measured by enzyme-linked immunosorbent assay(ELISA).The pathological changes of nasal and lung tissues were observed by hematoxylin-eosin(HE)staining.The expression of ILC2s in lung tissue of mice was detected by immunofluorescence(IF).The mRNA expression of GATA binding protein 3(GATA3),retinoic acid receptor-related orphan receptor-α(RORα),and inhibitor of DNA binding 2(ID2)in the lung tissue of mice was detected by quantitative real-time polymerase chain reaction(real-time PCR).The levels of IgE,interleukin(IL)-4,IL-5,and IL-13 in serum were detected by ELISA.Results:Compared with the blank group,the model group had poor survival state of mice and significantly increased serum IgE level after intranasal challenge(p<0.01).Additionally,the mice in the model group showed a large amount of neutrophil infiltration in the mucosa of the posterior turbinate,obvious nasal mucosal bleeding and purulent secretion,shed epithelium,thickened bronchial wall,obvious intravascular hyperemia and edema,diffusion and infiltration of a large number of inflammatory cells,seriously damaged alveolar structure,and local lung consolidation.The model group also exhibited significantly increased expression of ILC2s in the lung tissue(P<0.01),increased mRNA expression of GATA3 and RORα,decreased mRNA expression of ID2(P<0.05,P<0.01),and increased levels of serum IgE,IL-4,IL-5,and IL-13(P<0.05,P<0.01).Compared with the model group,the Mahuang Fuzi Xixintang group and the other medicine treatment groups showed improved survival state of mice,significantly reduced inflammatory cell infiltration in the nasal and lung tissues,a small amount of nasal mucosal bleeding,trachea wall thinning,and no hyperemia,edema,and nasal secretions.Furthermore,the expression of ILC2s in lung tissue was significantly decreased(P<0.01).The mRNA expression level of GATA3 was decreased(P<0.05),especially in the Aconiti Lateralis Radix Praeparata group(P<0.01).The expression mRNA levels of RORαwere decreased only in the Ephedrae Herba and Aconiti Lateralis Radix Praeparata group and the Ephedrae Herba group(P<0.05).The levels of serum IgE were decreased(P<0.05),and IL-5 levels were significantly decreased(P<0.01).IL-4 levels were significantly decreased in the groups except the Aconiti Lateralis Radix Praeparata group(P<0.01),and the level of IL-13 in the Mahuang Fuzi Xixintang group was decreased(P<0.05).The levels of IL-13 in were significantly decreased in the Ephedrae Herba and Aconiti Lateralis Radix Praeparata group,Ephedrae Herba group,Aconiti Lateralis Radix Praeparata group,and Asari Radix et Rhizoma group(P<0.01).Conclusion:Different compatibility of Ephedrae Herba,Asari Radix et Rhizoma,and Aconiti Lateralis Radix Praeparata can reduce the inflammation of OVA-induced AR mice and has more advantages in reducing the secretion of IgE and IL-5.The compatibility of Ephedrae Herba and Aconiti Lateralis Radix Praeparata has the most advantage in reducing the mRNA expression of GATA3 and RORαto inhibit the expression of ILC2s and thus exert the anti-allergic effect,while the other compatibility has the extensive advantage in inhibiting the mRNA expression of GATA3.
作者
张艺
陶晓华
刘敏
ZHANG Yi;TAO Xiaohua;LIU Min(Beijing University of Chinese Medicine,Beijing 100029,China)
出处
《中国实验方剂学杂志》
北大核心
2025年第11期51-59,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金面上项目(81774375)。
关键词
麻黄
细辛
附子
配伍
过敏性鼻炎
2型先天淋巴细胞(ILC2s)
小鼠
Ephedrae Herba
Asari Radix et Rhizoma
Aconiti Lateralis Radix Praeparata
compatibility
allergic rhinitis
type 2 innate lymphoid cells(ILC2s)
mice
