枸杞叶多糖提取工艺优化及其缓解小鼠过敏性鼻炎的研究

Optimization of extraction process of lycium barbarum leaf polysaccharide and its alleviation of allergic rhinitis in mice

为了优化枸杞叶多糖(lycium barbarum leaf polysaccharide,LBP)的提取工艺并探究LBP对小鼠过敏性鼻炎的免疫调节作用,构建卵清蛋白(ovalbumin,OVA)诱导的过敏性鼻炎模型,并给过敏性鼻炎小鼠灌胃优化工艺后得到的LBP。将30只雌性C57BL/6小鼠随机分为对照组,过敏性鼻炎模型组,LBP干预过敏性鼻炎组。从小鼠的常规状态,鼻黏膜组织化学染色以及脾脏Th1/Th2细胞比例3个方面评价LBP对小鼠过敏性鼻炎的调节作用,苏木精-伊红染色观察鼻黏膜的组织结构,过碘酸-希夫染色观察杯状细胞增生程度,甲苯胺蓝观察肥大细胞脱颗粒现象,髓过氧化物酶染色嗜酸性粒细胞的浸润程度。结果表明,枸杞叶多糖提取最佳条件为:提取时间8 h,温度50℃,料液比1∶25(g∶mL),LBP灌胃后的过敏性鼻炎小鼠常规状态改善,鼻黏膜炎症细胞的浸润程度减轻,脾脏Th1细胞比例升高,Th2细胞比例下降。

To optimize the extraction process of lycium barbarum leaf(LBP)and explore the immunomodulatory effect of LBP on allergic rhinitis in mice,an ovalbumin(OVA)induced allergic rhinitis model was constructed,and LBP was administered to mice with allergic rhinitis.Thirty female C57BL/6 mice were randomly divided into control group,allergic rhinitis model group and LBP intervention allergic rhinitis group.The regulatory effect of LBP on allergic rhinitis in mice was evaluated from three aspects:Routine status,nasal mucosal histochemical staining and Th1/Th2 cell ratio of spleen.Hematoxylin-eosin staining was used to observe the organizational structure of the nasal mucosa and periodic acid-Schiff staining was carried out to observe the degree of proliferation of goblet cells.Moreover,the nasal mucosa was stained with toluidine blue for observation of the degranulation phenomenon of mast cells and infiltration of eosinophils was observed by myeloperoxidase staining.The results showed that the optimal conditions for the extraction of LBP were as follows:The extraction was performed at 50℃for 8 hours with 1∶25 ratio of feed to liquid.After the treatment,the routine state of allergic rhinitis mice was improved after LBP gavage and the infiltration degree of inflammatory cells in nasal mucosa was reduced.Furthermore,the proportion of Th1 cells increased and Th2 cells were decreased in spleen.