黄芩汤提取物加减滴鼻液对实验性变应性鼻炎豚鼠IgE及肥大细胞的影响
摘要
变应性鼻炎(Allergic Rhinitis, AR),又称过敏性鼻炎,属中医学“鼻鼽”范畴。以发作性的鼻痒、鼻塞、喷嚏、流清水样鼻涕为主症,是发生在鼻粘膜的变态反应性疾病。本研究分上下两篇对黄芩汤提取物加减滴鼻液治疗实验性AR进行了文献综述和实验研究,通过制作AR豚鼠模型,观察黄芩汤提取物加减滴鼻液对实验性AR的疗效及作用机理,为临床提供科学依据。
上篇运用文献研究方法：
首先从中医学关于AR称谓的历史沿革、病因病机、辨证分型论治进行了全面综述。其次,介绍AR的西医研究现况,将西医对AR的认识、病因、发病机制、症状和体征、分类与分度、治疗概况进行了简要综述。最后,详细阐述了IgE、肥大细胞及TNF-a三者与AR的关系。
下篇实验研究包括：
1.方法
选取豚鼠60只,雌雄各半,按体重随机分成正常对照组、AR模型组、中药滴鼻液低频率组、中药滴鼻液中频率组、中药滴鼻液高频率组、富马酸酮替芬滴鼻液阳性对照组,每组10只。采用卵蛋白(OVA)对豚鼠基础致敏、雾化吸入强化致敏及局部滴鼻激发的方式制作AR模型。造模型成功后,分别对各组豚鼠给予生理盐水、黄芩汤提取物加减滴鼻液、富马酸酮替芬滴鼻液进行滴鼻,给药3周后处死豚鼠,采集血液与鼻粘膜组织标本,对以下项目进行观测。
(1)对各组动物进行行为症状学观察。
(2) ELISA法测定血清IgE的浓度。
(3) ELISA去测定血清TNF-α的含量。
(4)光学显微镜观察甲苯胺蓝染色法豚鼠鼻粘膜肥大细胞的浸润程度。
2结果
(1)行为症状学观察：模型组喷嚏次数和搔鼻次数均高于正常对照组,具有显着性差异(P<0.05)；各治疗组的喷嚏次数和搔鼻次数均低于模型组,具有显着性差异(P<0.05)。
(2)血清中IgE浓度：AR模型组IgE浓度显着高于正常对照组(P<0.01)；各治疗组均明显低于AR模型组(P<0.01)；中药中频组及高频组的血清IgE浓度与西药组的血清IgE浓度值在统计学上无显著性差异(P>0.05)。
(3)TNF-a含量：AR模型组血清TNF-a含量高于正常对照组(P<0.001)；各治疗组均明显低于模型组(P<0.001)。中药中频率及高频率组的血清TNF-a含量与西药组的血清IgE浓度值在统计学上无显著性差异(P>0.05)。
(4)肥大细胞：甲苯胺蓝染色光镜下正常组豚鼠鼻粘膜组织中可见肥大细胞散在分布,主要位于小血管和腺体周围。AR组豚鼠鼻粘膜组织中可见较多的肥大细胞在小血管和腺体周围成堆或成行的聚集。各治疗组豚鼠鼻粘膜组织中肥大细胞的浸润程度明显减少。
结论：黄芩汤提取物加减滴鼻液能够降低血清IgE浓度、稳定肥大细胞膜以抑制肥大细胞脱颗粒,调节TNF-a的含量,减弱致炎介质的产生,明显改善变应性鼻炎豚鼠的鼻部变态反应症状,具肯定疗效。
Allergic Rhinitis (AR), also known as rhinallergosis, belongs to the category of "Biqiu" in traditional Chinese medicine with the main symptoms of nasal itching, nasal obstruction, sneezing, watery nasal mucus and etc al. It is an allergic disease with the lesion located at the nasal mucosa. In the two parts of this study, we implemented the literature review and experimental study on the treatment of experimental AR using the modified decoction of Hangqintang nose drops. Through the AR cavy models, we observed the efficacy and action mechanism of the modified decoction of Hangqintang nose drops in the treatment of experimental AR in order to provide the scientific basis for clinical treatment.
In the first part of this study:
we reviewed in detail the history of AR appellation, and its pathogenesis as well as the planning treatment based on differentiation of syndrome and classification of types in traditional Chinese medicine. At the same time, we introduced the current research situation of AR in Western medicine and reviewed the understanding, etiology, pathogenesis, signs & symptoms, classification & indexing, and the treatment methods of the AR in Western medicine. Finally, we expounded the relationship of IgE, TNF-a and mast cells with AR.
The second part of this study includes:
1. Methods
60 guinea pigs, were randomly divided by weight into normal control group, AR model group, l(?)w-frequency Chinese medicine nasal drops group, moderate-frequency Chinese drug nasal drops group, high-frequency Chinese drug nasal drops group, and ketone fumarate nasal drops positive control group with 10 guinea pigs in each group.The AR models were constructed through basic sensitization using ovalbumin (OVA), enhanced sensitization by aerosol inhalation and local nose-dripping excitation. After successful modeling, cavies in each group were given intranasal treatment over 4 weeks with normal saline, modified decoction of Scutellaria Root nose drops, and ketone fumarate nasal drops respectively. At the end of 3rd week, all the guinea pigs were killed to collect the blood and nasal mucosa samples. Meanwhile, the following items were examined.
(1) behavior-semeiological observation before killing the animals in each group.
(2) measurement of the IgE concentration in serum.
(3) measurement of the TNF-a level in serum.
(4) toluidine blue staining observation of the nasal mucosa shape under optical microscope.
2. Results
(1) Behavior-semeiological observation:after four-week-treatment, the frequency of sneezing and scratching in model group was higher than that in normal control group with significant difference (P<0.05); the frequency of sneezing and scratching in treatment group was lower than that in normal control group with significant difference (P<0.05).
(2) IgE concentration in serum:the IgE concentration in the model group was significantly higher than that in the normal control group (P<0.01); meanwhile the IgE concentration in each of the treatment groups was significantly lower than that in the model group (P<0.01); however there was no significant difference between the Chinese medicine treatment groups and the western medicine treatment group (P> 0.05).
(3) TNF-a level:the TNF-a level in the model group was higher than that in the normal control group (P<0.001); meanwhile the TNF-a level in each of the treatment groups was significantly lower than that in the model group (P<0.001); however there was no significant difference between the Chinese medicine treatment groups and the western medicine treatment group (P> 0.05).
(4) Mast cells:the mast cells in nasal mucosa tissues from the normal control group was large and round with smooth edge and complete membrane; there were evenly stained brick-red particles with even distribution and uniform size in the cells which indicated that these were normal mucosal mast cells with non-stained nucleus. There were many degranulated mast cells in nasal mucosa tissues from the model group with broken membrane, escaped particles, swelling distorted deformed shape and unevenly stained particles. In the nasal mucosa tissues from the Chinese medicine treatment groups, there were a little number of degranulated mast cells. In the nasal mucosa tissues from the Western medicine treatment group, there were a small number of degranulated mast cells.
3. Conclusion
Because the modified decoction of Scutellaria root nose drops can reduce the IgE concentration and TNF-a level in serum, stabilize the mast cell membrane to inhibit mast cell degranulation, decrease the production of inflammatory mediators, and significantly improve the nasal allergic symptoms in allergic rhinitis cavies, therefore it has positive efficacy.
上篇运用文献研究方法：
首先从中医学关于AR称谓的历史沿革、病因病机、辨证分型论治进行了全面综述。其次,介绍AR的西医研究现况,将西医对AR的认识、病因、发病机制、症状和体征、分类与分度、治疗概况进行了简要综述。最后,详细阐述了IgE、肥大细胞及TNF-a三者与AR的关系。
下篇实验研究包括：
1.方法
选取豚鼠60只,雌雄各半,按体重随机分成正常对照组、AR模型组、中药滴鼻液低频率组、中药滴鼻液中频率组、中药滴鼻液高频率组、富马酸酮替芬滴鼻液阳性对照组,每组10只。采用卵蛋白(OVA)对豚鼠基础致敏、雾化吸入强化致敏及局部滴鼻激发的方式制作AR模型。造模型成功后,分别对各组豚鼠给予生理盐水、黄芩汤提取物加减滴鼻液、富马酸酮替芬滴鼻液进行滴鼻,给药3周后处死豚鼠,采集血液与鼻粘膜组织标本,对以下项目进行观测。
(1)对各组动物进行行为症状学观察。
(2) ELISA法测定血清IgE的浓度。
(3) ELISA去测定血清TNF-α的含量。
(4)光学显微镜观察甲苯胺蓝染色法豚鼠鼻粘膜肥大细胞的浸润程度。
2结果
(1)行为症状学观察：模型组喷嚏次数和搔鼻次数均高于正常对照组,具有显着性差异(P<0.05)；各治疗组的喷嚏次数和搔鼻次数均低于模型组,具有显着性差异(P<0.05)。
(2)血清中IgE浓度：AR模型组IgE浓度显着高于正常对照组(P<0.01)；各治疗组均明显低于AR模型组(P<0.01)；中药中频组及高频组的血清IgE浓度与西药组的血清IgE浓度值在统计学上无显著性差异(P>0.05)。
(3)TNF-a含量：AR模型组血清TNF-a含量高于正常对照组(P<0.001)；各治疗组均明显低于模型组(P<0.001)。中药中频率及高频率组的血清TNF-a含量与西药组的血清IgE浓度值在统计学上无显著性差异(P>0.05)。
(4)肥大细胞：甲苯胺蓝染色光镜下正常组豚鼠鼻粘膜组织中可见肥大细胞散在分布,主要位于小血管和腺体周围。AR组豚鼠鼻粘膜组织中可见较多的肥大细胞在小血管和腺体周围成堆或成行的聚集。各治疗组豚鼠鼻粘膜组织中肥大细胞的浸润程度明显减少。
结论：黄芩汤提取物加减滴鼻液能够降低血清IgE浓度、稳定肥大细胞膜以抑制肥大细胞脱颗粒,调节TNF-a的含量,减弱致炎介质的产生,明显改善变应性鼻炎豚鼠的鼻部变态反应症状,具肯定疗效。
Allergic Rhinitis (AR), also known as rhinallergosis, belongs to the category of "Biqiu" in traditional Chinese medicine with the main symptoms of nasal itching, nasal obstruction, sneezing, watery nasal mucus and etc al. It is an allergic disease with the lesion located at the nasal mucosa. In the two parts of this study, we implemented the literature review and experimental study on the treatment of experimental AR using the modified decoction of Hangqintang nose drops. Through the AR cavy models, we observed the efficacy and action mechanism of the modified decoction of Hangqintang nose drops in the treatment of experimental AR in order to provide the scientific basis for clinical treatment.
In the first part of this study:
we reviewed in detail the history of AR appellation, and its pathogenesis as well as the planning treatment based on differentiation of syndrome and classification of types in traditional Chinese medicine. At the same time, we introduced the current research situation of AR in Western medicine and reviewed the understanding, etiology, pathogenesis, signs & symptoms, classification & indexing, and the treatment methods of the AR in Western medicine. Finally, we expounded the relationship of IgE, TNF-a and mast cells with AR.
The second part of this study includes:
1. Methods
60 guinea pigs, were randomly divided by weight into normal control group, AR model group, l(?)w-frequency Chinese medicine nasal drops group, moderate-frequency Chinese drug nasal drops group, high-frequency Chinese drug nasal drops group, and ketone fumarate nasal drops positive control group with 10 guinea pigs in each group.The AR models were constructed through basic sensitization using ovalbumin (OVA), enhanced sensitization by aerosol inhalation and local nose-dripping excitation. After successful modeling, cavies in each group were given intranasal treatment over 4 weeks with normal saline, modified decoction of Scutellaria Root nose drops, and ketone fumarate nasal drops respectively. At the end of 3rd week, all the guinea pigs were killed to collect the blood and nasal mucosa samples. Meanwhile, the following items were examined.
(1) behavior-semeiological observation before killing the animals in each group.
(2) measurement of the IgE concentration in serum.
(3) measurement of the TNF-a level in serum.
(4) toluidine blue staining observation of the nasal mucosa shape under optical microscope.
2. Results
(1) Behavior-semeiological observation:after four-week-treatment, the frequency of sneezing and scratching in model group was higher than that in normal control group with significant difference (P<0.05); the frequency of sneezing and scratching in treatment group was lower than that in normal control group with significant difference (P<0.05).
(2) IgE concentration in serum:the IgE concentration in the model group was significantly higher than that in the normal control group (P<0.01); meanwhile the IgE concentration in each of the treatment groups was significantly lower than that in the model group (P<0.01); however there was no significant difference between the Chinese medicine treatment groups and the western medicine treatment group (P> 0.05).
(3) TNF-a level:the TNF-a level in the model group was higher than that in the normal control group (P<0.001); meanwhile the TNF-a level in each of the treatment groups was significantly lower than that in the model group (P<0.001); however there was no significant difference between the Chinese medicine treatment groups and the western medicine treatment group (P> 0.05).
(4) Mast cells:the mast cells in nasal mucosa tissues from the normal control group was large and round with smooth edge and complete membrane; there were evenly stained brick-red particles with even distribution and uniform size in the cells which indicated that these were normal mucosal mast cells with non-stained nucleus. There were many degranulated mast cells in nasal mucosa tissues from the model group with broken membrane, escaped particles, swelling distorted deformed shape and unevenly stained particles. In the nasal mucosa tissues from the Chinese medicine treatment groups, there were a little number of degranulated mast cells. In the nasal mucosa tissues from the Western medicine treatment group, there were a small number of degranulated mast cells.
3. Conclusion
Because the modified decoction of Scutellaria root nose drops can reduce the IgE concentration and TNF-a level in serum, stabilize the mast cell membrane to inhibit mast cell degranulation, decrease the production of inflammatory mediators, and significantly improve the nasal allergic symptoms in allergic rhinitis cavies, therefore it has positive efficacy.
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