摘要
研究目的：腧穴敷贴治疗变应性鼻炎因其在临床的广泛应用及确切的疗效,相关的效应及机制探讨已成为近年的研究热点,并取得了一定的研究成果。根据腧穴敷贴治疗变应性鼻炎计量学和Meta分析的结果发现,既往的研究虽然肯定了其临床疗效,但高质量的文献很少。机理研究有涉及AR发病机制中Th细胞亚群Th1/Th2的失衡关系,但并不深入,缺乏对近年免疫研究热点Th细胞另外两个亚群,Treg细胞和Th17的相关研究。目前临床使用的腧穴敷贴药物为中药散剂,存在药粉颗粒大小不一,很难混合均匀,影响贴敷效果的问题。因此,本研究拟从两个方面进行：一是采用细胞破壁,生物利用率高,能充分混匀且利于吸收的中药超微粉来进行腧穴敷贴,选取大椎、肺俞、肾俞穴治疗变应性鼻炎大鼠并观察其疗效；二是探寻超微粉治疗AR的可能机制,尤其是在既往研究的基础上深入探讨超微粉腧穴敷贴治疗对AR大鼠Th1、Th2、Treg和Th17细胞亚群的可能影响,以及细胞亚群之间的相互作用。
方法：造模与分组：全部56只SD大鼠,雄性28只,雌性28只,2月龄。分笼喂养,每笼6只。按随机数字表原则分成两组：正常对照组10只(雄5,雌5)；造模组46只(雄23,雌23)。适应性喂养3天后,造模组46只大鼠予卵清蛋白10mg+Al(OH)3粉末30mmg+生理盐水1m1制成混悬液腹腔内注射,隔日1次/d,共7次,作为基础致敏。第15天开始加强免疫,予5%OVA50u1喷鼻/(侧·日)和1%OVA喷雾吸入5min,连续7天。3周后,造模组46只大鼠中,有6只死于造模过程中(雄2只,雌4只),考虑为腹腔注射误注入大肠所致,剩余40只大鼠在末次激发后30min,1小时分别观察大鼠搔鼻、喷嚏、鼻溢等行为,并采用叠加计分法进行行为学评分,所有大鼠评分均超过5分,显示本组造模全部成功。将造模成功的AR模型大鼠再按随机数字表原则随机分为4组：模型组10只(雄6只,雌4只)；丙酸氟替卡松喷鼻剂组10只(雄5只,雌5只)；普通粉组10只(雄5只,雌5只)；超微粉组10只(雄5只,雌5只)。以上5组50只大鼠进入下一步实验。
穴位定位：大鼠大椎、肺俞、肾俞定位参考《常用实验动物穴位定位》。大椎位于第7颈椎与第1胸椎间,背部正中。肺俞位于第3胸椎下两旁肋间。肾俞位于第2腰椎下两旁。
普通粉以及超微粉的制备：普通粉、超微粉均选用符合《中华人民共和国药典》(2005版)中药饮片炮制规范,中药广州致信中药饮片有限公司生产,批号为080710。①普通粉：白芥子粉、延胡索粉、细辛粉、生甘遂粉,按照2：2：1：1的比例配置；②超微粉：将按比例配置好的普通粉由广州市第二中医院药剂科使用微粉机,振动电机功率为1.5FW,加工15分钟,粉碎成粒径范围0.1～75μm,粒度分布中心D50≈10～15μm,达300目以上的超微粉。
实验措施：①正常对照组：不采取任何处理措施；②模型组：给予生理盐水滴鼻,每日1次10天；③丙酸氟替卡松组：丙酸氟替卡松鼻喷雾剂喷鼻,每日1次,每次50gg/侧10天；④普通粉组：首先进行备皮,将按比例配置好的普通粉与新鲜姜汁调和均匀,制成约0.5cm0.5cm大小的药块,用3cm3cm次性纸胶布固定于穴位,再使用一次性透气孔胶布缠绕1-2圈以加固药贴。每次贴药4h10天；⑤超微粉组：贴敷药物改为超微粉,方法同上。
实验观察指标与检测：
①行为学观察：于末次治疗后30min、1小时观察大鼠搔鼻、喷嚏、鼻溢等行为的变化。记分方法：鼻痒：轻度搔痒1-2次为1分,剧烈搔痒抓鼻四周2分。喷嚏：1-3个为1分,4-10个为2分；11个以上为3分。清涕：流至鼻孔为1分,超出前鼻孔为2分,涕流满面为3分。采用叠加计分法。
②鼻黏膜组织HE染色及嗜酸性粒细胞计数观察：将各组大鼠于末次相关处理后,在10%水合氯醛,3ml/kg麻醉后,断头处死大鼠,迅速剥除上颌骨部皮肤,将上颌骨从颅骨中分离出来,打开鼻背,沿鼻中线切开,暴露鼻中隔和双侧鼻腔,分离鼻中隔与鼻黏膜,取双侧鼻黏膜。放入10%甲醛溶液中固定,石蜡包埋,常规切片行苏木精—伊红染色,光学显微镜下观测实验大鼠鼻黏膜组织学改变。每张病理切片随机选取5个高倍视野(1040)计数,每只大鼠鼻黏膜组织中嗜酸性粒细胞数作为一个计数。
③透射电子显微镜观察：鼻黏膜离体后必须在2分钟内,将标本置入2.5%戊二醛液中,4℃冰箱过夜。固定后的标本用磷酸缓冲液冲洗,并浸泡过夜。1%锇酸缓冲液固定1h,梯度乙醇脱水,Epon812包埋,聚合,超薄切片,醋酸铀、枸橼酸铅双重染色,用日立JEM1200-EX透射电子显微镜观察。
④免疫学指标测定：按照试剂盒说明采用酶联免疫法,检测血清中IL-4、TGF-β1、IFN-γ、IL-17的含量。
统计方法：行为学改变采用等级资料Kruskal-Wallis多个独立样本非参数检验,嗜酸性粒细胞计数及血清IFN-γ、IL-4,TGF-β1,IL-17采用单因素方差分析,多重比较采用LSD法,采用SPSS13.0统计分析,P<0.05为有统计学差异。
结果：行为学观察结果：①模型组与正常对照组在鼻痒、喷嚏、清涕等记分相比,分差显著,说明应用卵清蛋白致敏建立的AR动物模型成功；②由于非参数检验不能做多重比较,从平均秩次看三种治疗手段均有效,超微粉组从评分看治疗效果最好,丙酸氟替卡松次之,普通粉效果稍差,说明超微粉、普通粉和丙酸倍氯米松鼻喷雾剂均能明显改善变应性鼻炎的鼻痒、喷嚏、清涕等症状,使过变应性鼻炎大鼠症状得以大部或部分恢复。
嗜酸性粒细胞计数结果：模型组较正常对照组大幅升高,提示造模成功,超微粉组EOS计数较模型组及普通粉组显著下降(P<0.01),较丙酸氟替卡松组亦显著下降(P<0.05),与正常对照组相比无统计学差异(P=0.539),提示超微粉治疗大鼠变应性鼻炎局部疗效肯定；丙酸氟替卡松组及普通粉组EOS计数较模型组均明显下降(P<0.05),丙酸氟替卡松组EOS计数较普通粉组明显降低,有统计学差异(P<0.05),提示普通粉治疗变应性鼻炎效果不如鼻内激素。
HE染色图片分析表明：模型组与正常对照组比较炎症反应明显,毛细血管扩张、鼻黏膜上皮肿胀,渗透性增强,有大量嗜酸性粒细胞浸润固有层和血管周围,说明变应性鼻炎SD大鼠造模成功。药物干预的三组与模型组比较均很大程度的减轻了炎症反应,表现在上皮无明显肿胀,毛细血管扩张不明显,渗透性减少,以及无或者少量嗜酸性粒细胞浸润固有层和血管周围,说明药物干预组的治疗均达到预期的目标,可以缓解变应性鼻炎SD大鼠的鼻黏膜炎症反应。从三组图片结果看同EOS计数结果相似：丙酸氟替卡松组和超微粉组疗效优于普通粉组,但从HE染色的图片分析,丙酸氟替卡松组和超微粉组差异不大。
透射电子显微镜结果：模型组与正常对照组比较炎症反应明显,粘膜表面不完整,纤毛稀松畸形,见粗短纤毛、微绒毛,部分脱落,纤毛根部连接不完整,细胞内见肿胀线粒体及大小不等空泡,部分线粒体脊丢失,说明变应性鼻炎SD大鼠造模成功。药物干预的三组与模型组比较均有明显所改善,表现在粘膜表面基本完整,纤毛、微绒毛数量较模型组明显增多,细胞连接正常,超微粉组细胞内甚至可见正常内质网及线粒体,接近正常对照组的图片。说明药物干预组的治疗都达到预期的目标,可以缓解变应性鼻炎SD大鼠的鼻黏膜炎症反应。从三组图片结果看同EOS计数及HE染色结果相似：丙酸氟替卡松组和超微粉组疗效优于普通粉组,但与HE染色略微有差异的是超微粉组的透射电子显微镜图片可见细胞内正常内质网及线粒体,更接近于正常对照组的图片结果,疗效较丙酸氟替卡松组更好。
血清学观察结果：IFN-γ模型组较正常对照组大幅降低,提示造模成功,超微粉组IFN-γ较模型组、普通粉组及丙酸氟替卡松组显著升高(P<0.05),与正常对照组相比无统计学差异(P=0.584),丙酸氟替卡松组较模型组含量升高,有统计学差异(P<0.05),普通粉组较模型组相比含量升高,有统计学差异(P<0.05),而丙酸氟替卡松与普通粉之间无统计学差异(P=0.179),综合分析结果提示：超微粉、丙酸氟替卡松及普通粉治疗大鼠变应性鼻炎均显著升高了过敏导致的血清IFN-γ异常降低,同时超微粉效果最好,普通粉与鼻内激素效果相当。
IL-4模型组较正常对照组大幅升高,提示造模成功,超微粉组IL-4较模型组、普通粉组显著降低(P<0.05),与正常对照组及丙酸氟替卡松组相比无统计学差异(P=-0.095、0.07),丙酸氟替卡松组较模型组含量下降,有统计学差异(P<0.05),普通粉组较模型组相比含量下降,有统计学差异(P<0.05),而丙酸氟替卡松与普通粉之间无统计学差异(P=0.396),综合分析结果提示：超微粉、丙酸氟替卡松及普通粉治疗大鼠变应性鼻炎均显著降低了过敏导致的血清IL-4异常升高,同时超微粉与鼻内激素效果相当。
TGF-β1模型组较正常对照组大幅升高,提示造模成功,超微粉组TGF-β1较模型组、普通粉组及丙酸氟替卡松组显著下降(P<0.05),与正常对照组相比有统计学差异(P=0.05),丙酸氟替卡松组较模型组含量下降,有统计学差异(P<0.05),普通粉组较模型组相比含量下降,有统计学差异(P<0.05),而丙酸氟替卡松与普通粉之间无统计学差异(P=0.993),综合分析结果提示：超微粉、丙酸氟替卡松及普通粉治疗大鼠变应性鼻炎均显著降低了过敏导致的血清TGF-β1异常升高,同时超微粉效果最好,普通粉与鼻内激素效果相当。
IL-17模型组较正常对照组大幅升高,提示造模成功,超微粉组IL-17较模型组、普通粉组及丙酸氟替卡松组显著下降(P<0.05),与正常对照组相比无统计学差异(P=0.903),丙酸氟替卡松组较模型组含量下降,有统计学差异(P<0.05),普通粉组较模型组相比含量下降,有统计学差异(P<0.05),丙酸氟替卡松组较普通粉组IL-17升高,有统计学差异(P=0.01),综合分析结果提示：超微粉、丙酸氟替卡松及普通粉治疗大鼠变应性鼻炎均降低了过敏导致的血清IL-17异常升高,同时超微粉效果最好,鼻内激素次之,普通粉稍差。
结论：造模后AD大鼠行为学评分均超过5分,说明卵清蛋白腹腔注射是一种可靠地AR造模方法。综合EOS计数、组织HE染色、透射电镜及血清ELISA结果：超微粉、丙酸氟替卡松、普通粉组与模型组比均有显著差异,提示药物干预均能下调AR大鼠的炎症反应,有效治疗AR。尽管部分结果有差异,但总的趋势一致,显示超微粉疗效优于丙酸氟替卡松喷鼻剂及普通粉组,其可能机制：在药物及穴位刺激双重作用下,调整机体免疫状态,下调局部的炎症反应,同时Th细胞亚群在Treg细胞的核心调节作用下,使Th1/Th2失衡关系回归平衡,在促进Th17分化的同时,功能上又抑制了其促炎症反应,使整个精细而复杂的细胞因子网络在平衡与拮抗中发挥免疫功能。
OBJECTIVE:The crude herb moxibustion has been widely applied in treating allergic rhinitis in clinical practice which produces the exact effect. Its effectiveness and mechanism has become a research hotspot in recent years, and some achievements has been made. According to the metrology and Meta-analysis results of the application of crude herb moxibustion in treating allergic rhinitis, it is found that previous studies have confirmed its clinical efficacy though, literature of high quality are rare. Previous mechanism researches have touched upon the imbalance relationship in the Th1/Th2of Th cell subsets in AR pathogenesis though, they are not thorough and lack research related to two other subsets of Th cell, namely Treg cells and Th17, which are hotspots in recent immune researches. Currently, the acupoints application drugs for clinical use are tranditional Chinese medicine powder. There exist problem that powder particles in various sizes are difficult to mix evenly, undermining the sticking effect. Therefore, this study proceeds in two aspects. On one hand, it deploys the Chinese herbal submicron powder to destroy the cell wall, which is high bioavailability. The submicron powder used can be fully mixed, conducive to its absorption. The points of dazhui(DU14), feishu(BL13), shenshu(BL23) are selected to treat allergic rhinitis rats and the curative effect is observed. On the other hand, the study explores the possibility mechanism of submicron treatment of AR. Especially on the basis of previous studies, it approaches the therapy of the submicron powder acupoints application on the possible impact of AR rats with Th1, Th2, the subsets Treg and Th17cells in depth, in addition to the interaction between the cell subsets.
METHODS:Experimental animails and grouping:A total of56healthly SD rats aged of2months,28males,28females, were fed with six per cage. They were fractionated into two groups according to the random number table principles:10in control group (5male,5female);46in model group (23male,23female). After being adaptive fed for3days,46rats in made module are of intraperitoneal injection with suspension of ovalbumin10mg+Al (OH)3powder30mg+normal saline1ml every other day1/d and a total of7times, as a basis for sensitization. In the15days, we start to strengthen immunity with5%of OVA intranasally50ul/(side, d) and1%of OVA aerosol inhalation5min on consecutive7days. Three weeks later, among46rats in the model group,6died in the process of modeling (2male,4female), Which is considered to be caused by mistakenly intraperitoneal injection into the colon.30minutes after the last excitation, the remaining40rats were observed behavior of scratching nose, sneezing, rhinorrhea in an hour, and were used the superposition method of scoring for behavioral score. All rats'scores were more than five points, showing the model group was successful in all. Successful modeling of the AR model rats were randomly divided into4groups according to the principle of the random number table:10in model group (6male,4female);10in FP (Fluticasone propionate, FP) group (5male,5female);10in group of crude herb moxibustion (5male,5female);10in group of ultrafine herb moxibustion (5male and5female). Five groups of50rats mentioned were examined in the next experiments.
Location of Acupoints:The location of rats' points of dazhui(DU14), feishu(BL13), shenshu(BL23) is in accordance with The Acupoints Positioning of Commonly Used Experimental Animals. The point of dazhui is located between the7th cervical vertebra and the1st thoracic vertebra on the middle of the back. Feishu is below the3rd thoracic vertebra both sides of the intercostals space. Shenshu is located on both sides below the2nd lumbar vertebra.
Ordinary powder and submicron powder preparation:The selection of both crude herb and ultrafine herb powder was in line with the specification of Chinese herbal decoction pieces stated in The People's Republic of China Pharmacopoeia (2005edition). Chinese herbs were produced by Guangzhou Zhixin Chinese Herbal Pieces Co., Ltd. and the lot number was080710.①Crude herb powder:White mustard seed powder, corydalis tuber powder, asarum powder and raw euphorbia powder were configured in the2:2:1:1proportion.②Ultrafine herb powder:The ultrafine herb was more than300orders. That was used of micronized machine to crush the crude herbs powder in proportional allocation by the Second Traditional Chinese Medical Hospital of Guangzhou. The vibrating motor power was1.5FW in15minutes of processing. As a result, the powder was smashed into the size of0.1～75μm and particle size distribution center of D50≈10～15μm.
Experimental measures:①Normal control group:no measure was taken;②Model group:saline was given intranasally one time per day for10days;③The fluticasone propionate nasal spray group:the fluticasone propionate aerosol was used to consperge nose1times per day, each50μg/side10days;④The group of crude herb moxibustion(group A):First step was the skin preparation, prorata configured ordinary powder and fresh ginger juice were mixed about0.5cm x0.5cm size of the drugs block. Furthermore, to fix the points with the3cm3cm disposable paper tape, then disposable ventilating holes tape was used to wrap around1-2cycles in order to reinforce drug plaster. Each sticking drug was for4hours x10days;⑤The group of ultrafine herb moxibustion(group B):Application drugs was changed to submicron powder, and the method as above.
Experimental observation of targets and testing:
①Behavioral observation:After30min and1hour of the last treatment, we observed rats scratching nose, sneezing, experiencing rhinorrhea and other behavioral changes. Scoring method:Nasal itching:mild itching1-2times for1score, intense itching and scratch around the nose for2scores. Sneezing:1-3times for1score,4-10times for2scores, more than11times for3scores. Clear nasal discharge:Flowing to the nostrils for1point, beyond the anterior nostril for2points, and nasal discharge streamed all over the face for3points. We applied the method of superposition scoring.
②The nasal mucosa tissue HE staining and observation of eosinophil(EOS) count:After the last treatment, the rats in each group were anesthetized with3ml/kg10%chloral hydrate, then were decapitated to sacrifice. After that we quickly stripped the maxillary skin and separated the maxilla from the skull. Opened the nasal dorsum, and exposed the nasal septum as well as nasal cavity along nasal midline incision. And then, we separated nasal septum and nasal mucosa to take bilateral nasal mucosas which were fixed into10%formaldehyde solution and embedded in paraffin. The routine sections were in hematoxylin-eosin stain. We observed the histological changes of experimental rats'nasal mucosa under an optical microscope. Each of histologic sections randomly selected5high power fields (1040) to count and the number of eosinophils in the nasal mucosa of each rat was as a count.
③The TEM (Transmission electron microscope,TEM) observation:Within2minutes, the specimens of nasal mucosa in vitro should be placed in2.5%glutaraldehyde solution and4℃refrigerator overnight. The fixed specimens were flushed with phosphate buffer, and soaked overnight, then fixed1h with1%osmium tetroxide buffer solution and were gradient ethanol dehydration as well as Epon812embedding, polymerization, sliced ultrathin, with uranyl acetate and lead citrate double staining, finally, observed by Hitachi JEM1200-EX transmission electron microscope.
④The immunology index determination:using ELISA according to kit instructions to determine the content of IL-4、TGF-β1、IFN-γ、IL-17in serum.
Statistical analysis:The behavioral changes were processed with the Kruskal-Wallis of ranked data multiple independent samples of the nonparametric test. Eosinophil count and IFN-γ、IL-4, TGF-β1, IL-17in serum are utilized the single-factor variance analysis. The multiple comparisons were used the method of LSD and SPSS13.0statistical analysis. As result, P<0.05was considered statistically difference.
Results:
Behavioral observations:①There is significant difference between the scores of model group and the normal control group in nasal itching, sneezing and clear nasal discharge, indicating that AR animal models with ovalbumin sensitization is successfully established;②For the non-parametric tests can not do multiple comparisons, from the mean rank order method all of the three treatments are valid, from the score rating the group B get the best treatment, fluticasone propionate in turn, followed by group A less effective. It means that group B, group A, and beclomethasone dipropionate nasal spray could significantly improve or partly improve the symptoms of allergic rhinitis, including nasal itching, sneezing, clear nasal discharge, symptoms of allergic rhinitis in rats.
EOS count results:Compared with the normal control group, significant rise in the model group indicates the successful modeling. The EOS counting of submicron powder group is more significantly decreased (P<0.01) than the model group and group A. Compared with FP group there is also a significantly decrease (P<0.05); while compared with the control group, no significant difference (P=0.539) is observed, which indicates the confirmed local curative effect of group B on treating allergic rhinitis in rats; The EOS counting of FP group and normal powder group were significantly decreased than the model group(P<0.05), compared with the group A, the EOS counting of the FP group significantly decreased with statistical difference (P<0.05), indicating group A treatment of allergic rhinitis not as effective as the nasal hormone.
HE staining photographs analysis shows that:Compared with the control group, the model group has significant inflammatory response. angiotelectasis and nasal mucosal epithelium swelling, permeability increase, with a lot of eosinophilic infiltrated lamina propria and surrounding vessels, indicating allergic rhinitis SD rats modeling is successful. The three drug intervened groups compared with the model group are greatly reduced in the inflammation, showing no significant nasal mucosal epithelium swelling, no angiotelectasis and no or a small amount eosinophilic infiltrated lamina propria and surrounding vessels, indicating the treatments of all the drug intervened groups could achieve the desire objectives, which could release the nasal mucous membrane inflammation of allergic rhinitis SD rats. Results of the three groups'photographs are similar to the EOS counting results:FP group and group B were better than the normal group in group A, but HE staining photographs analysis shows no significant difference between the FP group and group B.
TEM observations:Inflammatory reaction obviously in the comparison between the model group and normal group, the mucosal surface is not complete, cilia thin and deformity, thick short cilia and microvilli could be seen, part of the cilia fall off, the cilia root incomplete connection in cells, swelling mitochondria and varying sizes vacuoles could be seen in the cells, part of the mitochondria ridge lost, indicating that allergic rhinitis SD rats modeling was successful. Three drug intervened groups achieve more significant improvement than the model group, showing in the mucosal surface is almost complete, the quantity of cilia and microvilli increased significantly, cell junction normal, normal endoplasmic reticulum and mitochondria could even be seen in cells of submicron powder group, which is close to the photograph of the normal control group. Showing that treatment of drug intervened group could reached the desired objective, and allergic rhinitis SD rats nasal mucous membrane of inflammation could be relieved. From the results we could see that the EOS counting is similar to that of HE staining:The curative effects of FP group and group B are better than that of the group A, while the slight difference of normal endoplasmic reticulum and mitochondria could be found in the transmission electron microscope images of group B, which is close to the photograph of the normal control group, better than FP group in curative effect.
Serologic observations:Compared with the normal control group, the serum levels of IFN-y model group is dramatically reduced, which indicates the modeling is successful. Compared with the group A and FP group, the serum levels of IFN-y of group B significantly increases (P<0.05), while there is no statistics difference with the comparison between group B and normal control group. Compared with the model group, the content of FP group increases with statistical difference (P<0.05).Compared with the model group, the content of the group A increases with statistical difference (P<0.05), while there is no statistics difference with the comparison between FP group and normal control group (P=0.179). All above shows that:group B, FP and group A treatment of allergic rhinitis of rats significantly increase the abnormal reduce allergy caused by serum IFN-y, with the group B having the best effect and the group A and FP almost the same.
The serum levels of IL-4model group compared with the normal control group significantly increases, indicating that the modeling was successful, the submicron powder of IL-4is significantly lower than that of model group, and the group A (P <0.05). Compared with FP group and the normal control group there is no statistically significant difference (P=0.095,0.07). Compared with the model group, content of the FP group decreaseds significantly (P<0.05), group A is statistically significant lower than that of model group (P<0.05), while no significant difference (P=0.396) between the FP with group A, a comprehensive analysis result suggests that:group B, FP for allergic rhinitis treatment in rats of Carcassonne and the group A significantly reduce the abnomal rise of causeserum IL-4caused by allergy, at the same time the similar effect can be found in the group B and FP.
Compared with the control group, the model group of TGF-β1increases significantly, suggesting that the modeling was successful. Compared with model group, group A and FP group, the TGF-β1of group B decreases significantly (P <0.05); compared with normal control group there is significant difference (P=0.05); content of the FP group decreases significantly (P<0.05) compared with that of the model group, content of group A compared with that of the model group decreased significantly as well (P<0.05); while there is no statistically significance between the FP group and the group A (P=0.993). a comprehensive analysis result suggests that: group B, FP and group A treatment of rats with allergic rhinitis were significantly reduced allergy caused by serum levels of TGF-β1abnormal rise, while the group B has the best effect, and group A and FP share similar effect.
Compared with control group, IL-17model group significantly increases, suggesting that the modeling is successful. Compared with model group, the group B, group A and FP group, IL-17of submicron powder group decreases significantly (P <0.05), and there is no significant difference compared to normal control group(P=0.903). Compared with model group, content of FP group decreased significantly (P <0.05), group A compared with the model group decreased significantly (P<0.05). FP group increases statistically significant (P=0.01) compared with IL-17of group A. A comprehensive analysis result suggests that:the group B, FP and group A treatment of allergic rhinitis in rats reduces the allergic abnormal rise as a result the application of serum IL-17. Also, group B has the best effect, which is followed by intranasal hormone and ordinary powder.
CONCLUSION:After the modeling, scores of the AD rat behavioral are all more than5, indicating that intraperitoneal injection ovalbumin (OVA) is reliable AR modeling method. Multiplied the EOS counting, the HE staining, transmission electron microscopy, and serum ELISA results:there were significant differences with the comparison of group B, FP, group A and model group, suggesting that the drug intervention can reduce inflammatory response of AR rats and it is an effective treatment of AR. Although some results are different, the overall trend is consistent with the fact that group B efficacy is superior to FP nasal spray agents and ordinary powder group. Its possible mechanisms are:the stimulations of both medicine and acupuncture points regulate the immune system and reduce the local inflammation. Meanwhile, with the regulatory role of Treg cells, Th cell subsets make the Th1/Th2imbalance relationship return to balance, which promotes Th17differentiation and inhibits proinflammatory function, so that the whole elaborate and complex cells factor networks plays in the balanced and antagonistic immune function.
引文
[1]邢志敏.变应性鼻炎的诊断和治疗进展[J].中国医学文摘(耳鼻咽喉科学),2010,25(4)：197-198
[2]王成硕.环境对变应性鼻炎的影响[J].中国医学文摘(耳鼻咽喉科学),2011,26(3)：152-155
[3]刘争,高起学,崔永华.美国变应性鼻炎最新调查报告[J].中国医学文摘耳鼻咽喉科学,2007,22(5)：289-291.
[4]Vichyanond P, Sunthornchart S, Singhirannusorn V, etal. Prevalence of asthma, allergic rhinitis and eczema among university students in Bangkok[J].Respir Med,2002,(2):34-38.
[5]韩德民,张罗,黄丹,等.我国11个城市变应性鼻炎自报患病率调查[J].中华耳鼻咽喉头颈外科杂志,2006,42(5)：378-384
[6]张罗,韩德民.变应性鼻炎治疗的进展[J].中华耳鼻咽喉头颈外科杂志,2005,40(3)：230-232.
[7]邓宁,罗永海.变应性鼻炎的中西医治疗现状[J].地方病通报,2009,24(6)：87-89.
[8]Members of the workshops. ARIA in the pharmacy:management of allergic rhinitis symptoms in the pharmacy. Allergic rhinitis and its impact on asthma[J]. Allergy,2004,59:373-387.
[9]Nagai H,Teramachi H,Tuchiya T.Recent advances in the development of anti-allergic drugs[J].Allergol Int,2006,55:35-42.
[10]李胜朴,刘小河.过敏性鼻炎的中医病因病机探讨[J].中外医疗,2008,17(3)：131-132.
[11]王德鉴,王士贞.中医耳鼻喉科[M].北京：人民卫生出版社.第版,2008：152-155.
[12]刘利民,严道南.变应性鼻炎中医治疗进展[J].江西中医药,2008,39(7)：75-76.
[13]谢水祥,韩立民.变应性鼻炎的病因病机及辨证论治[J].时珍国医国药,2010,21(5)：1280-1283.
[14]李寿今.干祖望治疗过敏性鼻炎的经验[J].湖北中医杂志,1981(3)：27.
[15]李国云,俞秋荣.中医药治疗变应性鼻炎的研究进展[J].湖北中医杂志,2010,32(8)：75-77.
[16]姜峰.过敏性鼻炎与中医体质的关系[J].中华中医药杂志,2008,23(2)：140-142.
[17]王士贞.中医耳鼻咽喉科学[M].北京：中国中医药出版社,2003：124-130.
[18]刘利民.变应性鼻炎的中医治疗进展[J].中医耳鼻喉科学研究杂志,2008,7(2)：34-36.
[19]王焱,马伟民.过敏性鼻炎的辨证论治[J].现代中医药,2004,1：35-37.
[20]陆寿康.刺法灸法学[M].北京：中国中医药出版社.2003：156.
[21]张晓明,庞巧玲.穴位贴敷法的作用机理[J].中国民间疗法,2005,13(8)：18-19.
[22]谢金城,谭寿如.穴位敷贴对过敏性鼻炎大鼠血清γ-干扰素和白介素-4含量的影响[J].中国中医基础医学杂志,2007,13(8)：614-615.
[23]许丽娟.变态反应性鼻炎的某些研究进展[J].中国中西医结合杂志,2002,10(1)：50.
[24]陈劫,赖新生,唐纯志,等.穴位敷贴对过敏性鼻炎小鼠腹腔肥大细胞脱颗粒影响的实验研究[J].上海针灸杂志,2007,26(12)：42-44.
[25]刘颖,潘德高,廖志航,等.七白散、七白微粉经皮给药的药效学对比研究[J].四川中医,2002,4(12)：13-15.
[26]苏艳丽,傅正义,马培艳,等.纳米化对三七中有效成分溶出的影响[J].中草药,2007,38(02)：202-204.
[27]杨一丁,王岩,高文勇,等.中药超微粉外用制剂治疗乳腺增生病的临床观察[J].中医外治杂志,2008,19(03)：30-31.
[28]侯慎英,刘兆琳.穴位贴敷法治疗变应性鼻炎168例总结[J].湖南中医杂志,2005,21(4)：23.
[29]毕冬桂.天灸疗法对变态反应性疾病治疗的临床疗效观察.中国临床医药研究杂志,2005,(144)：1591-1592.
[30]朱志康.也谈经络研究[J].上海针灸杂志,2001,20(1)：37-38.
[31]王雪苔.中国针灸大全[M].郑州：河南科学技术出版社.第一版,1992：6.
[32]黄泳,王升旭.针灸经络穴位速记手册[M].广州：广东科技出版社.第二版,2010：130-224.
[33]吴晖,庄金梅,李华,等.穴位贴敷治疗变应性鼻炎239例[J].福建中医学院学报,2005,15(5)：41-42.
[34]吕艳莹,李晓丽.穴位敷贴法配合穴位注射法治疗鼻鼽[J].内蒙古中医药,2009,28(19)：92-93.
[35]蔡恒,陈晓征.综合疗法治疗常年性变应性鼻炎40例[J].江西中医药,2006,37(7)：29-30.
[36]文志,徐绍勤,彭斌.穴位贴敷防治变应性鼻炎118例总结[J].湖南中医杂志,2007,23(2)：32-33.
[37]杜元灏,石学敏.中华针灸临床诊疗规范[S].南京：江苏科学技术出版社,2007：43.
[38]田道法.中西医结合耳鼻咽喉科学[M].北京：中国中医药出版社,2008：98.
[39]陈灏珠.实用内科学[M].北京：人民卫生出版社,第十二版,2005：2496.
[40]王雪峰,王加亮,董丹,等.小儿肺炎临床疗效观察类文献质量分析[J].中国实用儿科杂志,2007,22(4)：309-312.
[41]米建平,余焯粲.穴位贴敷法与激素吸入疗法治疗变应性鼻炎临床疗效对比分析[J].辽宁中医药大学学报,2011,13(1)：59-60.
[42]娄述,洪海国.针灸治疗过敏性鼻炎临床进展和思考[J].针灸临床杂志,2000,16(8)：57-59.
[43]中华医学会耳鼻咽喉科学分会,中华耳鼻咽喉科杂志编辑委员会.变应性鼻炎诊断标准及疗效评定标准[J].中华耳鼻咽喉科杂志,1998,33(3)：139.
[44]国家中医药管理局.中医病证诊断疗效标准[S].南京：南京大学出版社,1995：107.
[45]米建平,余焯粲.天灸治疗过敏性鼻炎临床观察[J].上海针灸杂志,2010,29(12)：773-775.
[46]刘素文,吴宏斌,张有忠.鹅蝥藿香粒天灸防治小儿变应性鼻炎93例临床观察[J].中医儿科杂志,2009,5(2)：44-45.
[47]孔亚明.天灸法治疗过敏性鼻炎的I临床观察[J].湖北中医杂志,2010,32(9)：49-50.
[48]张景琛.三伏天天灸疗法治疗小儿过敏性鼻炎的临床研究[D].广州：广州中医药大学2007届硕士学位论文,2007.
[49]林钺智.中药穴位贴敷治疗过敏性鼻炎临床疗效评估[D].南京：南京中医药大学硕士学位论文,2010.
[50]章如新,江德胜,周水淼,等.P物质能神经阻滞剂治疗变应性鼻炎的实验研究[J].中华耳鼻咽喉科杂志,1994,29(5)：282-285.
[51]Tanka K, OkamotoY. Anasal allergymodel developed in the guinea pig by intranasal application of- 2,4- toluene diisocyanate[J]. Int Archs Allergy Appl Immun,1988,85(4):392-397.
[52]张丽,杜经纬,王丽洪,等.豚鼠和新西兰兔变应性鼻炎模型建立及比较[J].重庆医科大学学报,2010,35(3)：335-338.
[53]赵宇,C.Andrew van Hasselt,吴港生,等.卵白蛋白经鼻致敏建立变应性鼻炎动物模型[J].中华耳鼻咽喉头颈外科杂志,2005,40(3)：176-180.
[54]杨瑞嘉,黄志纯.变应性鼻炎动物模型的建立及评价[J].国际耳鼻咽喉头颈外科杂志,2007,31(2)：87-89.
[55]邓可斌,王炼红.鼻鼽合剂对变应性鼻炎模型大鼠免疫系统的影响[J].湖北中医杂志,2011,33(6)：5-6.
[56]索利敏,赵长青,安云芳,等.变应性鼻炎大鼠鼻黏膜Bcl-2蛋白表达与细胞凋亡 的研究[J].中国药物与临床,2009,9(6)：479-481.
[57]李玉晓,陈冬,何晓光,等.大鼠变应性鼻炎模型中T- bet/GATA- 3基因表达的研究[J].昆明医学院学报,2011,32(4)：10-15.
[58]徐孝平,徐剑钦WHBE兔对卵白蛋白诱发变应性鼻炎的敏感性研究论[J].临床和实验医学杂志,2011,10(12)：881-885.
[59]张丽.甲苯-2,4二异氰酸酷经鼻致敏建立羊变应性鼻炎动物模型[D].重庆：重庆医科大学,2009.
[60]Tiniakov RL, Tiniakov OP, Mcleod RL, et al. Canine model of nasal congestion and allergic rhinitis [J]. J Appl Physiol,2003,94(5):1821-1828.
[61]冯纬纭,谢芳.变应性鼻炎动物模型造模综述[J].广西中医学院学报,2009,12(2)：77-79.
[62]Hunter DD, Satterfield BE, Huang J, et al. Toluene dissocyanate enhances substance P in sensory neurons innervating the nasal mucosa[J]. Am J Respi Crit Care Med,2000,161(2 Ptl):543-549.
[63]罗秋萍.硝酸铈对豚鼠变应性鼻炎鼻黏膜的影响[J].江西医药,2010,45(6)：530-531.
[64]席斌,田道法.变应性鼻炎肺气虚证与血清IL-4和IFN-r相关性的实验研究[J].中国中西医结合耳鼻咽喉科杂志,2007,15(3)：661-665.
[65]吉晓滨,邓家德,王磊,等.变应性算炎红细胞免疫功能的实验观察[J].中国耳鼻咽喉头颈外科,2006,13(11)：788-790.
[66]田理,贾德蓉,刘敬,等.力克敏喷剂治疗过敏性鼻炎实验研究[J].中医耳鼻喉科学研究杂志,2007,6(3)：28-30.
[67]慈军,王海英,沈国勤,等.尼多考米钠对实验性变应性鼻炎的疗效及对鼻孰膜P物质的影响[J].中国中西医结合耳鼻喉科杂志,2007,15(4)：248-251.
[68]陶峰,张凤兵,黄志纯,等.大鼠变应性鼻炎鼻私膜中脑源性神经营养因子mRNA的表达[J].临床耳鼻喉头颈外科杂志,2010,24(5)：224-226.
[69]许梅莉,郭永清.胸腺基质淋巴细胞生成素在变应性鼻炎模型小鼠鼻私膜中的表达[J].临床耳鼻喉头颈外科杂志,2009,23(17)：794-799.
[70]林静,魏永祥,王向东,等.变应性鼻炎伴嗅觉障碍小鼠嗅黏膜的观察[J].中国耳鼻咽喉头颈外科,2008,15(8)：465-468.
[71]胡艳菲,胡长庚,吴月琴,等.卵白蛋白诱发大鼠过敏性鼻炎模型不同方法的比较[J].吉林中医药,2011,31(10)：1023-1025.
[72]张枢,王宇,张宇,等.艾叶挥发油治疗大鼠变应性鼻炎的实验研究[J].中国免疫学杂志,2011,27(9)：787-789.
[73]李玉晓,陈冬,李添应,等.大鼠变应性鼻炎模型中T-bet和GATA-3的表达[J].临床耳鼻咽喉头颈外科杂志,2011,25(17)：791-795.
[74]罗灿娇,莫木琼,钟觉民,等.变应性鼻炎大鼠模型的建立方法[J].实用医技杂志,2008,15(29)：4020-4022.
[75]刘志刚,余洪猛,文三立,等.鹅不食草挥发油治疗过敏性鼻炎作用机理的研究[J].中国中药杂志,2005,30(4)：292-294.
[76]余洪猛,文三立,刘志刚,等.豚草花粉变应原致过敏性鼻炎豚鼠模型的建立[[J].上海实验动物科学,2001,21(4)：212-214.
[77]余洪猛,文三立,刘志刚,等.蛔虫变应原致过敏性鼻炎豚鼠模型的建立[J].上海实验动物科学,2000,20(4)：217-219.
[78]张志明,孙树岩,季文樾,等.真菌性抗原致变应性鼻炎豚鼠模型建立[J].中国耳鼻咽喉头颈外科,2005,12(5)：313-315.
[79]张鑫悦,卢良,刘利兵,等.蟑螂致变应性鼻炎豚鼠模型的建立[J].第四军医大学学报,2009,30(3)：279-281.
[80]国华,朱清仙,刘志刚,等.标准化粉尘螨疫苗鼻腔免疫治疗的疗效和机制研究[J].中华微生物学和免疫学杂志,2008,28(3)：273-274.
[81]林佳静,何晓华,胡雨华,等.三伏天穴位敷贴疗法对变应性鼻炎患者血清IgE、IL-4、IFN-γ的影响[J].宁夏医科大学学报,2011,33(11)：1013-1014.
[82]Romagnani S. Regulation of the T cell response [J]. Clin Exp Allergy,2006,36: 1357-1366.
[83]Castellino F, Germain R N. Cooperation between CD4+ and CD8+ T cells:when, where, and how [J]. Annu Rev Immunol,2006,24 (1):519-540.
[84]Park H, Li Z, Yang X O, et al. A distinct lineage of CD4+ T cells regulates tissue inflammation by producing interleukin 17[J]. Nat Immunol,2005,6 (11): 1133-1141.
[85]Harrington L E, Hatton R D, Mangan P R, et al. Interleukin 17-producing CD4+ effector T cells develop via a lineage distinct from the T helper type 1 and 2 lineages[J]. Nat Immunol,2005,6 (11):1123-1132.
[86]周光炎.免疫学原理[M].上海：上海科学技术出版社.2007：87
[87]Alves CR,Benevolo-De-Andrade, Alves JL,et al.Th1 and Th2 immunological profile induced by cysteine proteinase in murine leishmaniasis [J]. Parasite Immunol,2004,26(3):127-135.
[88]Bayry J, Sibe ril S, Triebel F, et al. Rescuing CD4+ CD25+ regulatory T-cell functions in rheumatoid arthritis by cytokine-targeted monoclonal antibody therapy [J]. Drug Discov Today,2007,12(13-14):548-552.
[89]Wahl S, Marten C, Mergenhagen S. Inflammatory and immunomodulatory roles TGF Beta 1 [J]. Immunol Today,1989,10:258-261.
[90]Gamble J, Vadas M. Enothelial adhesiveness for blood neutrophil is inhibited by transforming growth factor beta 1 [J]. Science,1998,242:97-99.
[91]MeCaffrey T, Baoheng D, Fu C, et al. The expression of TGF beta receptors in human atherosclerosis Evideneefoe acquired resistance to apoptosis due to receptor imbalance [J]. JMol Cell Cardiol,1999,31:1627-1642.
[92]Feiberg M, Jain M,Werner F, et al. Transforming growth factor beta 1 inhibits cytokine Gleizes PE, Rifk in DB. Activation of latent TGF-beta A required mechnism for vascular integrity [J]. Pathol Biol,1999,47(4):322-329.
[93]Miller R A. The aging immune system:primer and prospectus [J]. Science,1996, 273 (52717):70-74.
[94]Ruran M O,Jun C,Yin H et al. T cell chemokine receptor expression in aging [J]. Immunnology,2003,170(2):895-904.
[95]Muraille E,Leo O. Revisiting the Th1/Th2 paragigm [J]. Scand J Immunol,1998,47(1):129.
[96]Weiss S T. Eat dirt-the hygiene hypothesis and allergic disease [J]. New England J Medicine,2002,347(12):930.
[97]Romagnani S. Short analytical review:Thl and Th2 in human disease[J]. Clin Immunol Immunopathol,1996,80(3pt1):225-235.
[98]Prud homme GJ, Piceifillo CA. The inhibitory efects Of transforming growth factor-β1 (TGF-β1) in autoimmune diseases [J]. J Autoimmun,2000,14:23-42.
[99]Kim Y K,Myint A M,Lee B H et al. Th1,Th2 and Th3 cytokine alteration in schizophrenia [J]. Progress in Neuro-Psychopharmacology & Biological Psychiatry,2004,28 (1):1129-1134.
[100]Barbara G,Marzia C,Anna C et al.Th1,Th2 cytokines in the pathogenesis of bullous pemphigoid [J]. J Dermatological Science,2002,30 (2):116-128.
[101]Garder EM,Murasko DM. Age-releted changes in type1 and type2 cytokine production in humans[J]. Biogerontology,2002,3(5):271-289.
[102]Harrington L E, Mangan P R, Weaver C T. Expanding the effector CD4 T-cell repertoire:the Th17 lineage[J]. Curr Opin Immunol,2006,18(3):349-356.
[103]李佳,陈小平.CD4+效应T细胞的新成员-Th17细胞[J].中国免疫学杂志,2008,24(8)：763-766.
[104]Bettelli E, Korn T, Kuchroo VK. Th17:the third member of the effector Tcell trilogy[J]. Curr Opin Immunol,2007,19:1-6.
[105]葛伟,雷革非.Th17细胞的起源及其分化发育研究进展[J].现代免疫学,2011,31(6)：521-524.
[106]吴学忠.Th17细胞与TGF-β1相关性的研究进展[J].细胞与分子免疫学杂志,2008,24(1)：92-93.
[107]Annunziato F, Cosmi L, Santarlasci V, et al. Phenotypic and functional features of human Th17 cells[J]. J Exp Med,2007,204(8):1849-1861.
[108]Evans HG, Suddason T, Jackson I, et al. Optimal induction of T helper 17 cells in humans requires T cell receptor ligation in the context of Toll-like receptor activated monocytes[J]. Proc Natl Acad Sci USA,2007,104(43):17034-17039.
[109]Quinn DB, Mangan PR, Harrington LE, et al. Transforming growth factor-beta induces development of the Th17 lineage[J].Nature,2006,441(7090):231-234.
[110]Mc Innes IB, L iew FY.Cytokine networks-toward new therapies for rheumatoid arthritis[J].Nat Clin Pract Rheumatol,2005,1:31-39.
[111]卢汉桂,彭华,陈德华,等.转录因子RORC2和白细胞介素17的表达与变应性鼻炎症状关系的研究[J].中华耳鼻咽喉头颈外科杂志,2011,46(2)：144-147.
[112]唐碧霞.IL-17与自身免疫性疾病关系的研究进展[J].基础医学与临床,2008,28(1)：94-96.
[113]李鸿佳,王淑娟,董亮.Th17与慢性气道炎症性疾病[J].国际内科杂志,2007,34(10)：610-612.
[114]邢直直,杜文军,陈士俊,等.Th效应细胞家族的新成员-Th17[J].中国免疫杂志,2007,23：1048-1053.
[115]Ciprandi G, De Amici M, Murdaca G, et al. Serum interleukin-17 levels are related to clinical severity in allergic rhinitis[J]. Allergy,2009,64:1375-1378.
[116]Ated to clinical severity in allergic rhinitis[J].Allergy,2009,(64):1375-1378.
[117]Giorgio Ciprandi, Gilberto Filaci, etal.Peripheral Th-17 cells in allergic rhinitis: New evidence[J].International Immunopharmacology,2010,(10):226-229.
[118]Bousquet J,Cauwenberge P,Khaltaev N. Allergic rhinitis and its impact on asthma [J]. J Allergy Clin Immunol,2001,108(5):147-334.
[119]Umetsu DT,Dekruyff RH. Th1 and Th2 CD4+ cells in humanallergic diseases [J]. J Allergy Clin Immunol,1997,100 (1):1-6.
[120]刑同京,章廉.Th类细胞极化群体的基础与临床[M].北京：军事医学科学院出版社,2002,108-183.
[121]韩德民.变应性鼻炎[M].北京：人民卫生出版社.2007：263-265.
[122]李天春.变应性鼻炎的治疗研究进展[J].临床医药实践,2010,19(2)：230-232.
[123]李华斌,夏文彤ARIA指南2010版对抗组胺药物评价和使用建议[J].中国医学文摘耳鼻咽喉科学,2011,26(5)：247-249.
[124]赵宇.皮质类固醇在鼻炎治疗中的评价[J].中国医学文摘耳鼻咽喉科学,2010,25(4)：194-196.
[125]Mygind N. Local effect of intranasal beclomethasone dipropionateaerosol in hay fever[J]. Br Med J,1973,4:464-466.
[126]Bousquet J, Van Cauwenberge P, Khaltaev N. Allergic rhinitis and its impact on asthma[J]. J Allergy Clin Immunol,2001,108:147-334.
[127]Bousquet J,Khaltaev N,Cruz AA,et al. Allergic Rhinitis and its Impact on Asthma(ARIA)2008 update (in collaboration with the World Health Organization,GA(2)LEN and AllerGen. Allergy,2008,63:8-160.
[128]中华耳鼻咽喉头颈外科杂志编委会,中华医学会耳鼻咽喉科学分会.变应性鼻炎的诊治原则和推荐方案(2004年,兰州).中华耳鼻咽喉头颈外科志,2005,40：166-167.
[129]中华耳鼻咽喉头颈外科杂志编委会鼻科组,中华医学会耳鼻咽喉科学分会鼻科学组.变应性鼻炎的诊治原则和推荐方案(2009年,武夷山).中华耳鼻咽喉头颈外杂志,2009,44：977-978.
[130]Waddell AN, Patel SK, Toma AG, et al. Intranasal steroid sprays in the treatment of rhinitis:is one better than another? [J]. Laryngol Otol,2003,117:843-845.
[131]Scadding GK. Intranasal steroid sprays in the treatment of rhinitis is one better than another? [J].Laryngol Otol,2004,118:395-396.
[132]Van Adelsberg J, Philip G, Pedinoff J, et al. Montelukast improves symptoms of seasonal allergic rhinitis over a 4-week treatment period[J].Allergy,2003, 58:1268-1276.
[133]李志鹏,王丹凤.克敏通窍止嚏汤对变应性鼻炎大鼠IgE、IL-4和IL-5含量的影响[J].浙江中西医结合杂志,2010,20(4)：212-214.
[134]周林,郑鸣Toll样受体4和NF-2κB p50在大鼠变应性鼻炎中的表[J].四川解剖学杂志,2010,18(1)：3-5.
[135]赵义造,黄益灯,黄加云,等.电针对变应性鼻炎大鼠模型IL-4及IFN-y表达的影响[J].浙江中医杂志,2010,45(12)：916-918.
[136]刘志严.小青龙汤对变应性鼻炎大鼠IL-4和IFN-y含量的影响[J].辽宁中医药大学学报,2011,13(10)：249-251.
[137]罗一鸣,黄志纯,孙宝宾.抗神经生长因子抗体对变应性鼻炎大鼠鼻黏膜炎症的影响[J].东南大学学报,2011,30(3)：495-499.
[138]谢冰斌,刘月辉,朱新华,等.丙酸氟替卡松对转化生长因子β1蛋白及其mRNA在大鼠变应性鼻炎鼻钻膜中表达的影响[J].中国耳鼻咽喉头颈外科,2011,18(7)：382-384.
[139]刘建国,刘月辉,张海珍,等.丙酸氟替卡松对实验性变应性鼻炎鼻黏膜血红素氧合酶-1的影响[J].临床耳鼻咽喉头颈外科杂志,2012,26(2)：78-81.
[140]谢冰斌,刘月辉,刘建国,等.丙酸氟替卡松对变应性鼻炎鼻黏膜重塑动态影响的实验研究[J].临床耳鼻咽喉头颈外科杂志,2011,25(5)：232-235.
[141]冯宝民,余正江,李帆.白芥子化学成分的研究[J].大连大学学报,2004,26(4)：43-45.
[142]刘兴隆,贾波,黄秀深,等.细辛药理研究概况[J].江苏中医药,2005,26(7)：59-61.
[143]程建明,井山林.延胡索中延胡索乙素不同提取方法的比较研究[J].时珍国医国药,2006,17(7)：1236-1237.
[144]王玉波,李颖玉,王红兵,等.甘遂的化学成分[J].中国天然药物,2007,5(3)：182-185.
[145]孙静,陶欣,朱镇华.穴位贴敷法治疗变应性鼻炎临床观察[J].湖南中医药大学学报,2007,27(3)：63-64.
[146]吴晖,庄金梅,李华,等.穴位贴敷治疗变应性鼻炎239例[J].福建中医学院学报,2005,15(5)：41-42.
[147]侯慎英,刘兆琳.穴位贴敷法治疗变应性鼻炎168例总结[J].湖南中医杂志,2005,21(4)：23.
[148]程倩,陈协云.穴位贴敷法治疗变应性鼻炎的研究进展[J].湖南中医药大学学报,2007,27(1)：75-79.
[149]徐珊宁,陈苏,徐芸.穴位敷贴治疗过敏性鼻炎30例[J].福建中医药,2005,36(3)：37.
[150]杨连威,赵晓燕,李婷,等.中药超微粉碎后对其性能的影响研究[J].世界科学技术-中医药现代化,2008,10(6)：77-80.
[151]李德成,刘庆燕.超微粉碎技术在中药制剂中广泛应用的优越性[J].世界中医药,2011,6(5)：454-455.
[152]刘大伟,武文斌.浅谈中药超微粉碎技术的现状与发展[J].黑龙江医药,2010,2(5)：763-765.
[153]方邦江,陈浩,郭全,等.中药超微粉的优势及应用前景[J].中国中医药现代远程教育,2010,18(8)：208-209.
[154]欧阳荣,胡铁骊,凌可.当归超微细粉、浓缩颗粒与饮片水提取液主成分含量的对比研究[J].中国中医药信息杂志,2004,11(12)：1071-1072.
[155]李虹,张文高.西洋参微粉对老年冠心病病人免疫功能的影响[J].中西医结合 心脑血管病杂志,2005,3(10)：847-849.
[156]徐英杰,尹羽薇,董清平.中药超微粉贴敷干预对兔骨性关节炎关节软骨中MMP-1、TIMP-1表达影响的实验研究[J].中医药信息,2011,28(4)：134-136.
[157]刘迎,史万玉,钟秀会.微粉中药促兔散对小鼠迟发型变态反应的影响[J].中兽医医药杂志,2009,4(4)：13-15.
[158]史万玉,包永占,林德贵,等.观察中药黄藿散微粉对免疫抑制小鼠非特异性免疫功能的影响[J].中国兽医杂志,2007,43(12)：45-46.
[159]陈开文,谭涌.中药超微粉碎应用研究概况[J].中国药业,2006,15(2)：75-77.
[160]张素萍,胡能.超微粉碎技术在中药生产中应用探讨[J].贵州化工,2008,33(3)：29-32.
[161]朱莉,隆泉,郑保忠.超微粉碎技术及其在中药加工中的应用[J].云南大学学报(自然科学版),2004,26(增刊)：130.
[162]靳子明,贺沙沙.中药超微粉碎技术的优势及存在的问题[J].甘肃中医学院学报,2011,28(5)：77-79.
[163]孙文格.中医药应用超微粉碎技术的研究进展[J].中国药业,2009,18(20)：80-81.
[164]李先端,程立平,仝燕,等.祛除白附子麻辣刺激性新技术-超微粉碎[J].中国实验方剂学杂志,2008,14(9)：26-29.
[165]胡连生,李凡成.中医耳鼻咽喉科学[M].北京：中国中医药出版社.2004：186-187.
[166]朱玉琴.过敏性鼻炎当责之肺脾肾[J].河南中医,2000,20(6)：16.
[167]黄桂锋.培土生金法治疗脾气虚变应性鼻炎大鼠的实验研究[D].成都：成都中医药大学,2010：31-34.
[168]徐轩.略谈肾阳虚与变态反应性鼻炎[J].南京中医学院学报,1988,4：35.
[169]陈劫,杨挺,赖新生,等.穴位敷贴对过敏性鼻炎小鼠血细胞分类和血清SIgE的影响[J].广州中医药大学学报,2008,25(7)：304-306.
[170]Tada T, Okumura K L. Regulation of homocytotropic antibody Formation in the rat III.Effect of thymectomy and splenectomy[J]. Immunol,1971,106:1019.
[171]Lee I H. Cytokine network in the pathogenesis of bronch ial asthmaimplication for therapy [J].JR Coll Physicians Lond,1998,32:56.
[172]谢金城,谭寿如.穴位敷贴对过敏性鼻炎大鼠血清γ-干扰素和白介素-4含量的影响[J].中国中医基础医学杂志,2007,13(8)：614-615.
[173]许丽娟.变态反应性鼻炎的某些研究进展[J].中国中西医结合杂志,2002,10(1)：50.
[174]陈劼,赖新生,唐纯志,等.穴位敷贴对过敏性鼻炎小鼠腹腔肥大细胞脱颗粒影响的实验研究[J].上海针灸杂志,2007,26(12)：42-44.
[175]Furuno T, Hirashima N, Onizawa S, et al. Nuckear shuttling of mitogen-activated protein (MAp) kinase extracellular signal-regulated kinase (ERK) was dynamically controlled byMAp/ERK kinase after antigen stimulation in RBL-2H3 cells [J]. Immunol,2001,166:4416-4421.
[176]赖新生,贾钰华.针灸对I型变态反应疾病血浆前列腺素含量的影响[J].针灸学报,1992,(1)：4-6.
[177]顾之燕,顾瑞金.鼻分泌物介质研究进展[J].中华耳鼻咽喉科杂志1989,24(2)：112.
[178]张乃元.针刺对过敏性鼻炎大鼠免疫调节作用机制的研究[D].广州：广州中医药大学,2011.
[179]金弘.针刺五脏俞治疗围绝经期综合征临床观察[J].中国针灸,2007,27(8)：572-574.
[180]臧江红,王丽华,郭裕.三伏天穴位敷贴治疗过敏性鼻炎疗效观察[J].中医学报,2012,27(2)：245-246.
[181]许凤丽,周艳霞,李建超,等.穴位贴敷治疗发作期过敏性鼻炎的临床报道[J].针灸临床杂志,2011,27(7)：46-47.
[182]陈兴华,文庆华,徐涵斌.三伏天穴位贴敷治疗过敏性鼻炎116例临床观察[J].江苏中医药,2011,43(1)：64-65.
[183]夏寅,朱丽相.变应性鼻炎嗜酸性粒细胞研究进展[J].国外医学耳鼻喉分册,1996,20(2)：72-75.
[184]黄桂锋,卢健敏,熊大经,等.中药复方对变态反应性鼻炎大鼠鼻黏膜组织Eotaxin-mRNA表达的影响[J].长春中医药大学学报,2011,27(6)：912-914.
[185]蔡文琴.现代实用细胞与分子生物学实验技术[M].北京,人民军医出版社.2003：223.
[186]杨春平,罗英,刘月辉Th17/Treg细胞与变应性鼻炎的研究进展[J].重庆医学,2011,40(26)：2685-2687.
[187]席斌,田道法.变应性鼻炎肺气虚证与血清IL-4和IFN-y相关性的实验研究[J].中国中西医结合耳鼻咽喉科杂志,2007,15(3)：166-170.
[188]刘志严.小青龙汤对变应性鼻炎大鼠IL-4和IFN-y含量的影响[J].辽宁中医药大学学报,2011,10(10)：249-251.
[189]赵义造,黄益灯,黄加云,等.电针对变应性鼻炎大鼠模型IL-4及IFN-y表达的影响[J].浙江中医杂志,2010,45(12)：916-918.
[190]Veldhoen M, H ocking RJ, Flavel 1 RA, et al. Signals medated by transforming growth fact or beta initiate autoimmune encephalom yelit is, but chronic inflammat-ion is needed to sustain disease[J]. Nat Immunol,2006,7(11) 1151-1156.
[191]张蓝方Th17/Treg平衡调节与疾病的关系研究进展[J].济宁医学院学报,2011,34(1)：68-70.
[192]沈暘,洪苏玲,胡国华,等.鼻息肉患者外周血Th17/Treg细胞比率的失衡及其临床意义[J].细胞与分子免疫学杂志,2011,27(12)：1139-1141.
[193]张成,洪苏玲,胡国华.变应性鼻炎患者Treg/Th17细胞相关转录因子及细胞因子的失衡表达[J].临床耳鼻咽喉头颈外科杂志,2012,26(5)：209-211.
[194]Blobe GC, Schiemann WP, Lodish HF. Role of transforming growth factor beta in human disease[J]. N Engl J Med,2000,342:1350-1358.
[195]Kenyon NJ, Ward RW, McGrew G, et al. TGF-betal causes airway fibrosis and increased collagen I and III mRNA in mice[J].Thorax,2003,58:772-777.
[196]江满杰,李泽卿,吴晋蓉,等.抗原长期刺激致变应性鼻炎豚鼠模型鼻黏膜重塑的研究[J].临床耳鼻咽喉头颈外科杂志,2012,26(5)：218-222.
[197]Nakamura T, Sakata R, Ueno T, et al. Inhibition of transforming growth factor beta prevents progression of liver fibrosis and enhances hepatocyte regeneration in dimethylnitrosamine-treated rats[J]. Hepatology,2000,32:247-255.
[198]Miyamoto M, Prausc O, Sjostrand M, et al. Endogenous IL-17 as a mediator of neutrophil recruitment caused by endotoxin exposure in mouse airways [J]. J Immunol,2003,170(9):4665-4672.
[199]Zou GM, Tam YK. Cytokines in the generation and maturation of dendritic cells:recent advances[J].European Cytokine Net,2002,13(2):186-199.
[200]Ciprandi G, De Amici M, Murdaca G, et al. Serum interleukin-17 levels are related to clinical severity in allergic rhinitis[J]. Allergy,2009,64(9):1375-1378.
[201]Cheung PF, Wong CK, Lam C.W. Molecular mechanisms of cytokine and chemokine release from eosinophils activated by IL-17A, IL-17F, and IL-23: implication for Th17 lymphocytesmediated allergic inflammation [J]. Immunol, 2008,180:5625-5635.
[202]李忠仁.腧穴配伍研究概况[J].中国针灸,2010,30(5)：437-439.