玉屏风散对变应性鼻炎模型小鼠肥大细胞成熟及活化的影响

Effect of Yupingfeng Powder on the Maturation and Activation of Mast Cells in Mice with Allergic Rhinitis

目的研究玉屏风散对变应性鼻炎(AR)模型小鼠肥大细胞成熟及活化的影响。方法30只雄性BALB/c近交系小鼠按随机数字表法分为正常对照组、AR模型组和玉屏风散组,每组10只。AR模型组及玉屏风散组通过卵清蛋白(OVA)联合Al(OH)_(3)腹腔注射及OVA滴鼻激发鼻黏膜法建立AR模型,玉屏风散组于激发前1天开始用玉屏风散水煎剂灌胃(25g·kg^(-1)·d^(-1))治疗,连续8天,正常对照组和AR模型组则用等量生理盐水灌胃。采用鼻部症状评估小鼠AR症状;酶联免疫吸附试验(ELISA)检测血浆组胺和白介素-13(IL-13)水平;甲苯胺蓝染色检测鼻黏膜肥大细胞浸润;免疫组织化学染色检测鼻黏膜类胰蛋白酶的表达;流式细胞仪检测骨髓细胞中表面标志物特异性受体(CD117)、FcεRI表达。结果与AR模型组比较,玉屏风散组小鼠的挠鼻、喷嚏及流清涕评分和鼻部症状总评分显著降低[(1.40±0.70)分比(2.60±0.52)分,(1.60±0.70)分比(2.80±0.42)分,(1.50±0.71)分比(2.50±0.53)分,(4.50±1.51)分比(7.90±1.20)分,P均<0.05];血浆组胺和IL-13水平显著降低[(6.58±0.83)ng/mL比(12.58±1.16)ng/mL,(46.27±6.17)pg/mL比(63.38±5.02)pg/mL,P均<0.05];鼻黏膜组织每高倍镜下肥大细胞数量及类胰蛋白酶表达水平显著降低[(12.33±4.50)个比(20.67±8.33)个,(0.23±0.08)比(0.35±0.17),P均<0.05];骨髓细胞CD117和Fc着RI双阳性细胞比例显著降低[(4.95±0.81)%比(8.83±0.92)%,P<0.05]。结论玉屏风散可抑制肥大细胞分化、成熟,降低其脱颗粒及炎性因子分泌水平,从而减轻AR模型小鼠的鼻部症状。

Objective To determine the effect of Yupingfeng powder on the maturation and activation of mast cells in a mouse model of allergic rhinitis(AR).Methods Thirty male BALB/c inbred mice were divided into control,AR model,and AR model plus Yupingfeng powder treatment groups according to the random number table(n=10 mice per group).The AR model was produced by using ovalbumin(OVA)nasal drip plus Al(OH)_(3) intraperitoneal injection.Yupingfeng powder was administrated in the Yupingfeng group of mice one day before the model was introduced through oral gavage(25g·kg^(-1)·d^(-1))for eight consecutive days.Both control groups of mice(real and AR model controls)were given the same amount of normal saline.Nasal symptom score was used to evaluate AR symptoms in mice,and the ELISA was used to assess plasma levels of histamine and interleukin 13(IL-13).The toluidine blue staining was used to detect nasal mucosal mast cell infiltration,immunohistochemistry was used to detect the expression of nasal mucosal tryptase,and flow cytometry was applied to detect the expression of surface markers CD117 and FcεRI in bone marrow cells.Results Compared with the AR model group,scores of the nasal scratching,sneezing,and runny nose as well as the total score of nasal symptoms of mice were significantly reduced in the Yupingfeng powder group of mice(1.40±0.70 vs.2.60±0.52,1.60±0.70 vs.2.80±0.42,1.50±0.71 vs.2.50±0.53,and 4.50±1.51 vs.7.90±1.20,respectively;P<0.05).The plasma levels of histamine and IL-13 were also significantly reduced(6.58±0.83 vs.12.58±1.16 and 46.27±6.17 vs.63.38±5.02pg/mL,respectively;P<0.05),while the mast cell number and level of tryptase expression in nasal mucosa tissues were significantly downregulated in treatment mice(12.33±4.50 vs.20.67±8.33 and 0.23±0.08 vs.0.35±0.17,respectively;P<0.05).Moreover,the ratio of CD117 and FcεRI double-positive cells in the bone marrow cells was significantly reduced in treatment mice(4.95±0.81 vs.8.83±0.92%,P<0.05).Conclusion The Yupingfeng power was able to inhibit mast cell differentiation and maturation as well as reduce mast cell degranulation and inflammatory cytokine secretion;thereby,reducing the nasal symptoms in mice with allergic rhinitis.