白芥子贴膏穴位贴敷对过敏性鼻炎大鼠的免疫调制作用研究

Immunomodulation of Acupoint Application with Semen Sinapis Plaster on Rats with Allergic Rhinitis

[目的]探讨白芥子贴膏穴位贴敷对过敏性鼻炎大鼠的疗效以及对免疫调制的影响。[方法]将50只Wistar大鼠随机分为对照组、模型组、西药组、生白芥子组、炒白芥子组,每组10只。除对照组外,其余各组大鼠均应用卵白蛋白建立过敏性鼻炎模型,造模成功后西药组用丙酸氟替卡松喷鼻,每侧鼻孔5μL;生白芥子组和炒白芥子组分别用生白芥子贴膏和炒白芥子贴膏贴敷;模型组用0.9%氯化钠溶液滴鼻,每侧鼻孔5μL。各组均隔日治疗1次,共治疗7次。干预结束后观察各组大鼠治疗前后行为学表现,以酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)法测定大鼠血清中白介素-1β(interleukin-1β,IL-1β)、白介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和干扰素-γ(interferon-γ,IFN-γ)表达水平,并探讨几个指标之间的关系;对各组大鼠鼻黏膜组织细胞进行甲苯胺蓝法染色,镜下观察鼻黏膜组织肥大细胞形态并计数。[结果]与对照组比较,模型组大鼠过敏性鼻炎总症状评分显著增高(P<0.05);与模型组比较,各治疗组总症状评分显著降低(P<0.05);与西药组比较,生白芥子组治疗后评分显著降低(P<0.05),炒白芥子组差异无统计学意义(P>0.05)。与对照组比较,模型组血清中IL-1β、IL-6、TNF-α水平显著升高(P<0.05),IFN-γ水平显著降低(P<0.05);与模型组比较,各治疗组大鼠血清IL-1β、IL-6、TNF-α水平显著降低(P<0.05),IFN-γ水平显著升高(P<0.05);与西药组比较,治疗后生白芥子组IL-1β、IL-6、TNF-α水平显著下降,IFN-γ水平显著上升(P<0.05);炒白芥子组IL-1β、IL-6、TNF-α和IFN-γ水平差异无统计学意义(P>0.05)。Pearson相关性分析提示,大鼠血清IL-1β和IL-6、TNF-α的水平呈正相关(r=0.841、r=0.595,均P<0.05);和IFN-γ的水平呈负相关(r=-0.818,P<0.05);IL-6和TNF-α的水平呈正相关(r=0.69,P<0.05);和IFN-γ的水平呈负相关(r=-0.861,P<0.05);TNF-α和IFN-γ的水平呈负相关(r=-0.68,P<0.05)。与对照组比较,病理染色显示模型组大鼠鼻黏膜组织肥大细胞显著增多(P<0.05);与模型组比较,各治疗组鼻黏膜肥大细胞数量明显减少(P<0.05);与西药组比较,生白芥子组肥大细胞计数明显降低(P<0.05),炒白芥子组差异无统计学意义(P>0.05)。[结论]生/炒白芥子贴膏穴位贴敷均可有效治疗过敏性鼻炎,而且生白芥子贴膏效果更为显著,其机制可能与降低血清IL-1β、IL-6、TNF-α水平,升高IFN-γ水平,改善鼻黏膜炎性症状相关。

[Objective]To explore the curative effect of acupoint application of raw and fried Semen sinapis plaster on rats with allergic rhinitis(AR)and the related mechanism of immunomodulation.[Methods]Using random number table method,fifty Wistar rats were randomly divided into control group,model group,western medicine group,raw Semen sinapis group and fried Semen sinapis group,10 in each group.Except for control group,all the other groups of rats used oval bumin to build AR model.After successful modeling,rats in western medicine group were administrated with fluticasone propionate,5μL in each nostril;rats in raw Semen sinapis group and fried Semen sinapis group used raw/fried Semen sinapis plaster,and rats in model group were administrated with 0.9%sodium chloride solution,5μL in each nostril,each group was treated once every other day,for a total of 7 treatments.The behavior of each group was observed before and after treatment.The serum levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ)were determinated by enzyme linked immunosorbent assay(ELISA)and the relationship between the indexes was explored.The nasal mucosa tissue of each group was stained with toluidine blue method,and the number of mast cells in the nasal mucosa tissue was observed and counted.[Results]Compared with control group,the AR total symptom scores of model group were significantly higher(P<0.05);compared with model group,the total scores of each treatment group were significantly lower(P<0.05);compared with western medicine group,the scores of raw Semen sinapis group were significantly lower(P<0.05),but there was no significant difference in fried Semen sinapis group(P>0.05).Compared with control group,serum levels of IL-1β,IL-6,TNF-αin model group were significantly higher(P<0.05),while levels of IFN-γwere significantly lower(P<0.05);compared with model group,the serum levels of IL-1β,IL-6 and TNF-αin treatment groups were significantly lower(P<0.05),but the levels of IFN-γwere significantly higher(P<0.05);compared with western medicine group,the levels of IL-1β,IL-6,TNF-αin raw Semen sinapis group significantly decreased,and the level of IFN-γsignificantly increased(P<0.05);there was no significant difference in the levels of IL-1β,IL-6,TNF-αand IFN-γin fried Semen sinapis group(P>0.05).Pearson correlation analysis showed that serum levels of IL-1βwere positively correlated with IL-6 and TNF-α(r=0.841,r=0.595,all P<0.05),negatively correlated with IFN-γ(r=-0.818,P<0.05);the level of IL-6 was positively correlated with TNF-α(r=0.69,P<0.05),negatively correlated with IFN-γ(r=-0.861,P<0.05);and the level of TNF-αwas negatively correlated with IFN-γ(r=-0.68,P<0.05).Compared with control group,pathological staining showed that the number of mast cells in nasal mucosa tissue of model group was significantly higher(P<0.05);compared with model group,the number of mast cells in nasal mucosa tissue intreatment groups significantly reduced(P<0.05).Compared with western medicine group,the mast cell count in raw Semen sinapis group was significantly lower(P<0.05),but there was no significant difference in fried Semen sinapis group(P>0.05).[Conclusion]Acupoint application of raw/fried Semen sinapis plaster can effectively treat AR,and the effect of raw Semen sinapis plaster is more significant.The mechanism may be related to lowering the serum levels of IL-1β,IL-6,TNF-αwhile increasing the level of IFN-γ,and improving inflammatory response of nasal mucosa.