补气通窍方灌胃对大鼠变应性鼻炎的治疗作用及其机制

Therapeutic effect of intragastric administration of Buqi Tongqiao formula on allergic rhinitis in rats and its mechanism

目的通过建立大鼠变应性鼻炎(AR)模型,观察补气通窍方对大鼠AR的治疗作用并分析其机制。方法将Wistar大鼠随机分为对照组、模型组、补气通窍方组及鼻炎康组,每组10只。除对照组外均使用卵清蛋白抗原佐剂混悬液建造AR模型,建模成功后,模型组给予4 mL/d纯净水灌胃,补气通窍方组给予补气通窍方57.55g/(kg·d)灌胃,鼻炎康组给予鼻炎康462.5 mg/(kg·d)灌胃,对照组常规饲养不灌胃,共灌胃7 d,每日1次。各组每次灌胃结束后观察大鼠30 min内抓鼻、喷嚏及流涕的症状发生情况并计算动物行为学积分;HE染色观察鼻黏膜组织病理学改变;Western blotting法检测脾脏组织NF-κB p65蛋白;16 S rRNA基因测序检测肠道菌群多样性及丰度。结果模型组、补气通窍方组及鼻炎康组各时点动物行为学积分均高于对照组;灌胃2 d后,补气通窍方组动物行为学积分均低于模型组;灌胃3 d后,鼻炎康组动物行为学积分均低于模型组(P均<0.05)。对照组鼻黏膜假复层纤毛柱状上皮清晰可见,固有层无水肿、充血等病理改变;模型组鼻黏膜纤毛排列杂乱,可见较多嗜酸性粒细胞等炎症细胞浸润,腺体数量增加;补气通窍方组及鼻炎康组固有层黏膜较为完整,炎症细胞浸润程度较模型组减少,无明显腺体增生,鼻黏膜病理情况好转。脾脏组织NF-κB p65蛋白表达模型组>补气通窍方组、鼻炎康组>对照组(P均<0.05)。对照组、模型组、补气通窍方组及鼻炎康组分别包含3687、1641、1222、891种菌群,各组大鼠两两比较,组间菌群结构存在差异。在菌属水平上,各组大鼠的肠道菌群特定菌属具有显著差异(P均<0.05)。在科水平上,乳杆菌科相对丰度对照组>补气通窍方组、鼻炎康组>模型组(P均<0.05)。结论补气通窍方可改善大鼠AR的鼻炎症状,修复鼻黏膜组织病理损伤,其机制可能与提高乳杆菌相对丰度调节肠道微生态,下调脾脏组织NF-κB p65蛋白表达有关。

Objective By establishing rat models of allergic rhinitis(AR),the therapeutic effect of Buqi Tongqiao formula(BQTQF)on AR rats was observed and its mechanism was analyzed.Methods Wistar rats were randomly divided into four groups:control group,model group,BQTQF group,and Biyankang group,with 10 rats in each group.Except the control group,AR models were established with the suspension of ovalbumin antigen adjuvant in the other groups.After modeling,rats in the the model group were administered intragastrically 4 mL/d pure water,rats in the BQTQF group were administered 57.55 g/(kg·d)of BQTQF intragastrically,and rats in the Biyankang group were given 462.5 mg/(kg·d)of Biyankang intragastrically,while rats in the control group were fed routinely without gavage.The symptoms of nasal scratching,sneezing and runny nose were observed within 30 min after the end of gavage and the ethological scores were calculated;the pathological changes of nasal mucosa in rats with AR were observed by HE staining;Western blotting was used to detect NF-κB p65 protein in spleen;16S rRNA gene sequencing was used to detect the diversity and abundance of intestinal flora.Results The scores of animal behavior in the model group,BQTQF group and Biyankang group were higher than those in the control group.After 2 days of intragastric administration,the scores of BQTQF group were lower than those of the model group.After 3 days of intragastric administration,the scores of Biyankang group were lower than those of the model group(all P<0.05).In the control group,there were clear nasal mucosa pseudostratified ciliated columnar epithelium,but there were no pathological changes such as edema and congestion in the lamina propria.In the model group,the nasal mucocilia were disordered,and more inflammatory cells such as eosinophils were infiltrated and the number of glands increased.The lamina propria mucosae of BQTQF group and Biyankang group were relatively intact,the degree of inflammatory cell infiltration was reduced in comparison with that of the model group;there was no obvious glandular hyperplasia,and the pathological condition of nasal mucosa was improved.Spleen NF-κB p65 protein expression was as follows:model group>BQTQF group,Biyankang group>control group(all P<0.05).The control group,model group,BQTQF group and Biyankang group contained 3687,1641,1222 and 891 species of microflora,and there were differences in microflora structure between groups.At the level of bacterial genus,there were significant differences in the specific bacterial genera of intestinal flora among all groups.At the family level,relative abundance of Lactobacillaceae was in the following order:control group>BQTQF group,Biyankang group>model group(P<0.05).Conclusion BQTQF can improve the symptoms of rhinitis and repair the pathological injury of nasal mucosa in AR rats,and its mechanism may be related to increasing the relative abundance of Lactobacillaceae,regulating intestinal microecology,and down-regulating the expression of NF-κB P65 protein in spleen.