银翘散对鼻黏膜屏障功能低下模型小鼠的影响

Improvement Effect of Yinqiao Powder on the Model Mice with Low Nasal Mucosal Barrier Function

目的研究银翘散对模型小鼠鼻黏膜屏障功能低下的改善作用。方法实验分为正常对照组及模型A,B组,模型A,B组小鼠分别予脂多糖(LPS,2 mg/kg)单次或连续多次滴鼻,正常对照组小鼠予等体积磷酸盐缓冲液滴鼻;于LPS单次滴鼻后第1,3,5,7天,以及连续滴鼻(每天1次;1,3,5,7次)后24 h时,采用Western blot法检测小鼠鼻黏膜中免疫球蛋白A(IgA)和紧密连接蛋白(Occludin)的表达水平;于第7次滴鼻前1 h以异硫氰酸荧光素(FITC)标记的葡聚糖滴鼻,采用酶标仪测定小鼠血清中葡聚糖水平。实验分为正常对照组(等体积蒸馏水)、模型C组(等体积蒸馏水)及银翘散低、高剂量组(生药24.18,72.54 g/kg),均于LPS滴鼻后6 h灌胃相应药物(0.2 mL/10 g)或蒸馏水,每天1次,连续7 d,检测小鼠鼻黏膜中IgA和Occludin蛋白表达水平及血清葡聚糖水平。结果LPS单次滴鼻,与正常对照组比较,模型A组小鼠鼻黏膜组织中IgA和Occludin蛋白表达水平仅分别在滴鼻后第1天和第3天时显著降低(P<0.05);LPS多次滴鼻,与正常对照组比较,模型B组小鼠连续滴鼻1,3次时的鼻黏膜组织中IgA蛋白表达水平显著降低,连续滴鼻3,5,7次时的Occludin蛋白表达水平显著降低及血清葡聚糖水平显著升高(P<0.05)。与正常对照组比较,模型C组小鼠鼻黏膜组织中Occludin蛋白表达水平显著降低,血清葡聚糖水平显著升高。与模型C组比较,银翘散低、高剂量组小鼠鼻黏膜组织中Occludin蛋白表达水平显著升高,血清葡聚糖水平显著降低;银翘散高剂量组小鼠鼻黏膜组织中IgA蛋白表达水平显著升高(P<0.05)。结论银翘散可改善模型小鼠鼻黏膜免疫屏障功能,机制为促进鼻黏膜Occludin蛋白表达,降低鼻黏膜上皮屏障的通透性,从而提高鼻黏膜机械屏障的功能。还可通过促进IgA的表达,提高鼻黏膜免屏障功能。

Objective To study the improvement effect of Yinqiao Powder on the model mice with low nasal mucosal barrier function.Methods The experiment was divided into the normal control group and the model groups A and B.The mice in the model groups A and B were given single and continuous multiple nasal drops of lipopolysaccharide(LPS,2 mg/kg)respectively,and the mice in the normal control group were given an equal volume of phosphate buffer nasal drops.The expression levels of immunoglobulin(IgA)and Occludin protein in the nasal mucosa of mice were detected by the Western blot on the first,third,fifth and seventh days after administration of single nasal drip of LPS and at the 24 h after continuous nasal drip of LPS(once a day,1,5,7 times).Fluorescein isothiocyanate(FITC)-dextran was used for nasal drip at 1 h before the seventh nasal drip,and the serum dextran level in mice was determined by the enzyme-linked immunosorbent assay(ELISA).The experiment was divided into the normal control group(equal volume of distilled water),model group C(equal volume of distilled water)and Yinqiao Powder low-and high-dose groups(24.18,72.54 g crude drug/kg),with eight mice in each group.The corresponding drugs(0.2 mL/10 g)or equal volume of distilled water were administered by gavage at 6 h after nasal drip of LPS,once a day for 7 d.The expression levels of IgA and Occludin protein in nasal mucosa and the serum dextran level were detected.Results Compared with those in the normal control group,the expression levels of IgA and Occludin protein in the nasal mucosa of mice in the model group A were significantly lower only on the first and third days after single nasal drip of LPS(P<0.05).Compared with those in the normal control group,the expression level of IgA protein in the nasal mucosa of mice was significantly lower after continuous nasal drip of LPS for 1 and 3 times in the model group B,the expression level of Occludin protein was significantly lower and the level of serum dextran was significantly higher after continuous nasal drip of LPS for 3,5,7 times in the model group B(P<0.05).Compared with those in the normal control group,the expression level of Occludin protein in the nasal mucosa was significantly lower,while the level of serum dextran was significantly higher in model group C.Compared with that in the model group C,the expression level of Occludin protein in the nasal mucosa of mice in Yinqiao Powder low-and high-dose groups was significantly higher,the level of serum dextran in Yinqiao Powder low-and high-dose groups was significantly lower,while the expression level of IgA protein in the nasal mucosa of mice in high-dose Yinqiao Powder group was significantly higher(P<0.05).Conclusion Yinqiao Powder can improve the function of immune barrier of nasal mucosal in model mice,its mechanism relates to promoting the expression level of Occludin protein in nasal mucosa,reducing the permeability of nasal mucosal epithelial barrier,so as to improve the mechanical barrier function of nasal mucosa.It can also improve the immune barrier function of nasal mucosa by promoting the expression of IgA.