针刺对过敏性鼻炎大鼠免疫调节作用机制的研究
摘要
一、研究目的
过敏性鼻炎(Allergic Rhinitis, AR)也称变态反应性鼻炎、变应性鼻炎,是-种吸入变应原而导致的以鼻黏膜嗜酸性粒细胞、肥大细胞、T淋巴细胞等多种炎性细胞浸润,IgE介导的免疫细胞释放化学介质引起的Ⅰ型变态反应炎症性疾病,以鼻塞、鼻痒、打喷嚏、流清涕等为主要症状,可并发哮喘、荨麻疹等。有国外研究表明过敏性鼻炎发生、发展与社会开放程度和经济发展水平密切相关；近几年,我国经济发展很快,过敏性鼻炎的发病率也明显呈上升趋势,我国该病发病率在10%左右,对人民身心健康造成严重影响。
现代医学治疗过敏性鼻炎以免疫疗法和药物治疗为主,两种疗法均存在不足。免疫疗法,变应原较难确定而且标准难一致,起效慢,疗程长,不易坚持,难以治愈；药物治疗以抗组胺药、缩血管药、糖皮质激素、抗胆碱能药、肥大细胞稳定剂等为主,虽然改善症状较快,但毒副作用大。
祖国医学中过敏性鼻炎属于“鼻鼽”、“鼽嚏”的范畴。针刺疗法治疗过敏性鼻炎在古籍中早有记载,而且也取得较好的疗效。虽然针刺疗法治疗过敏性鼻炎取得较好疗效,但现代中医学研究大都处于临床观察或个别免疫指标检测等较浅层次阶段,对其作用机制的研究报道则较少。本论文系统研究针刺疗法治疗大鼠过敏性鼻炎的作用机制,为针刺疗法治疗过敏性鼻炎提供科学的理论依据。
二、研究方法与内容
(一)文献研究
运用文献研究方法,分别阐述祖国医学古代医家、现代医学、现代中医学对过敏性鼻炎的临床与实验研究,并在此基础上进行评述与展望。
(二)实验研究
本研究主要通过不同处理组别对过敏性鼻炎大鼠行为学、病理学、鼻腔灌洗液白三烯C4、外周血IL-4、鼻黏膜Toll样受体4和NF-κB蛋白表达等多个环节的影响,研究针刺疗法治疗过敏性鼻炎的机理,从细胞、分子水平揭示针刺疗法治疗过敏性鼻炎的机制。
1.实验动物与分组：SPF级健康SD大鼠40只,雌雄各半,体重180-220g。随机分为4组,即模型组、针刺组、西药组、空白组,每组10只；
2.实验动物造模：以卵蛋白注射及其鼻粘膜刺激法对针刺组、西药组、模型组建立变应性鼻炎动物模型；
3.干预措施：针刺组选取大椎、肺俞(双)、肾俞(双)穴进行针刺治疗,西药组以辅舒良气雾剂喷鼻治疗,模型组给予生理盐水滴鼻刺激,空白组不做处理；
4.观察不同处理组别卵蛋白所致过敏性鼻炎大鼠模型行为学及病理组织学改变；
5.观察不同处理组别卵蛋白所致过敏性鼻炎大鼠模型鼻腔灌洗液中白三烯的改变；
6.观察不同处理组别卵蛋白所致过敏性鼻炎大鼠模型大鼠外周血IL-4的改变；
7.观察不同处理组别卵蛋白所致过敏性鼻炎大鼠模型大鼠鼻黏膜TOll样受体4和NF-κB蛋白表达的改变；
8.分析各相关指标的改变意义,阐明针刺治疗过敏性鼻炎的免疫学作用机制。
三、研究结果
1.与空白组比较,模型组大鼠鼻痒、喷嚏和流清鼻涕等行为学方面积分显著增多(p<0.05),经过针刺治疗和辅舒良气雾剂喷鼻治疗后行为学积分与模型组比较有明显降低(p<0.05)
2.与空白组比较,模型组大鼠鼻粘膜嗜酸性粒细胞计数明显增高(p<0.05),经过针刺治疗和辅舒良气雾剂喷鼻治疗后鼻粘膜嗜酸性粒细胞计数与模型组比较有明显降低(p<0.05)
3.与空白组比较,模型组大鼠鼻腔灌洗液中LTC4含量明显升高(p<0.05),经过针刺治疗和辅舒良气雾剂喷鼻治疗后大鼠鼻腔灌洗液中LTC4含量与模型组比较有明显降低(p<0.05),西药组优于针刺组。
4、与空白组比较,模型组大鼠血清IL-4含量明显升高(p<0.05),经过针刺治疗和辅舒良气雾剂喷鼻治疗后大鼠血清IL-4含量与模型组比较有明显降低(p<0.05)。
5.与空白组比较,模型组大鼠鼻黏膜Toll样受体4蛋白表达平均光密度和NF-κB蛋白表达平均光密度明显升高(p<0.05),经过针刺治疗和辅舒良气雾剂喷鼻治疗后大鼠鼻黏膜Toll样受体4蛋白表达平均光密度和NF-κB蛋白表达平均光密度与模型组比较有明显降低(p<0.05)。
四、结论
1.针刺激肾俞(双)、肺俞(双)、大椎对过敏性鼻炎大鼠鼻痒、喷嚏、清涕等症状的改善有明显效果,即对过敏性鼻炎有较好的治疗效果,这进一步证实了针灸通过调理肺肾、升提阳气治疗过敏性鼻炎的正确性和合理性。
2.针刺治疗过敏性鼻炎大鼠的机理
(1)针刺治疗在一定程度上可以抑制炎症细胞,特别是嗜酸性细胞在气道内的聚集和活化,抑制嗜酸性细胞的活力,起到治疗气道炎症的作用。
(2)针刺可以降低过敏性鼻炎鼻腔灌洗液中LTC4的含量、降低血清中IL-4的含量及鼻黏膜TLR4和NF-κB蛋白表达,提示针刺对过敏性鼻炎的治疗效果是通过作用于多个环节而达到其治疗效果,特别是在对Toll-NF-κB信号通路上进行多层次调节。
Objective
Allergic rhinitis(AR) is allergenically inflammatory disease of type I induced by allergen, which is by The method of IgE and is characteristic by The esophocyte, mast cells, T lymphocyte. The symptom is nasal itch, sneeze and snivel with The complication of asthma. The abroad study revealed the occur of allergenic nasatitis is closely related to economic development. Recent years, the mobidity have inceased significantly to 10% with serious damage to the health of PeoPle.
AR is treated by modern medicine with immunal Therapy or durg, which have a lot of shortcoming. For immunal Therapy, The allergenic antigen is difficult to b3:determined, The effect is slow to occur, The course is too long to finish for the patients, The disease is difficult cured. The drug treating Allergenic nasatitis consist of durg anti-histamine, cortisol, atropine, with rapid effect and more side effect.
AR is calle "biqiu", "qiuti" in Traditional Chinese Medicine. Acupuncture treats the AR with long time and good results. But the immunal mechanism is undermined. In our study, the immunal mechanism of Acupuncture treating the allergenic nasatitis in investigated, which Provide scientific (?)rmation for therapy. Methods
1. Published Articles are Studied
The Published artieles are sudtied to declare the clinical and experimental investigation and provide the prospection.
2. Experimental Study
(?) our study, the effects of Acupuncture on The AR are investigation in (?)vation, pathology, TLC4 in NLF, IL-4 in blood, TLR4 and NF-κB in Nose mucosal.
2.140 SD rats, SPF and 180-220g weight are randomly divided into four groups:normal control group, model group, acupuncture treatment group and Fluticasone Propionate Nasal Spray(FPNS) treatment group.
2.2 Rat model of allergic rhinitis was prepared by injecting egg album and stimulating nasal mucosa.
2.3 Normal control group was not treated. acupuncture treatment group was treated by acupuncture on DU 14(Dazhui), BL 23(Shenshu) and BL 13(Feishu). FPNS group was treated by FPNS, model group was treated by physiological saline.
2.4 The behaviors of rats in all groups were observed. The pathological changes of nasal mucosa were tested and the contents of TLC4 in NLF, IL-4 in blood, TLR4 and NF-κB in Nose mucosal were determined by radio-immunologic analysis.
Results
1. Compared with normal control group, the behaviors'scores of model group were significantly higher (p<0.05). After treatment, the behaviors' scores of acupuncture treatment group and FPNS group were significantly lower than those of model group (p<0.05)
2. Compared with normal control group, the eosinophilic granulocyte(EOS) count of model group is significantly higher (p<0.05). After treatment, the EOS counts of acupuncture treatment group and FPNS group were significantly lower than those of model group (p<0.05)
3. Compared with normal control group, the serum content of TLC4 in NLF and IL-4 in blood of model group is significantly higher (p<0.05). After treatment, the serum content of TLC4 in NLF and IL-4 in blood of acupuncture treatment group and FPNS group were significantly lower than those of model group (p<0.05)
4. Compared with normal control group, the expression of TLR4 and NF-κB of model group is significantly higher (p<0.05). After treatment, the expression of TLR4 and NF-κB of acupuncture treatment group and FPNS group were significantly lower than those of model group (p<0.05)
Conclusion
1. Acupuncture treatment can effectively improve the symptoms of rats with allergic rhinitis. It means the method is effective for treating allergic rhinitis.
2. The possible action mechanisms of Acupuncture treatment in treating allergic rhinitis may be as the follows::①It can significantly decrease the EOS count of nasal mucosa of rats with allergic rhinitis.②It can effectively decrease the serum content of TLC4 in NLF and IL-4 in blood of rats with allergic rhinitis.③It can effectively decrease the expression of TLR4 and NF-κB of rats with allergic rhinitis.
过敏性鼻炎(Allergic Rhinitis, AR)也称变态反应性鼻炎、变应性鼻炎,是-种吸入变应原而导致的以鼻黏膜嗜酸性粒细胞、肥大细胞、T淋巴细胞等多种炎性细胞浸润,IgE介导的免疫细胞释放化学介质引起的Ⅰ型变态反应炎症性疾病,以鼻塞、鼻痒、打喷嚏、流清涕等为主要症状,可并发哮喘、荨麻疹等。有国外研究表明过敏性鼻炎发生、发展与社会开放程度和经济发展水平密切相关；近几年,我国经济发展很快,过敏性鼻炎的发病率也明显呈上升趋势,我国该病发病率在10%左右,对人民身心健康造成严重影响。
现代医学治疗过敏性鼻炎以免疫疗法和药物治疗为主,两种疗法均存在不足。免疫疗法,变应原较难确定而且标准难一致,起效慢,疗程长,不易坚持,难以治愈；药物治疗以抗组胺药、缩血管药、糖皮质激素、抗胆碱能药、肥大细胞稳定剂等为主,虽然改善症状较快,但毒副作用大。
祖国医学中过敏性鼻炎属于“鼻鼽”、“鼽嚏”的范畴。针刺疗法治疗过敏性鼻炎在古籍中早有记载,而且也取得较好的疗效。虽然针刺疗法治疗过敏性鼻炎取得较好疗效,但现代中医学研究大都处于临床观察或个别免疫指标检测等较浅层次阶段,对其作用机制的研究报道则较少。本论文系统研究针刺疗法治疗大鼠过敏性鼻炎的作用机制,为针刺疗法治疗过敏性鼻炎提供科学的理论依据。
二、研究方法与内容
(一)文献研究
运用文献研究方法,分别阐述祖国医学古代医家、现代医学、现代中医学对过敏性鼻炎的临床与实验研究,并在此基础上进行评述与展望。
(二)实验研究
本研究主要通过不同处理组别对过敏性鼻炎大鼠行为学、病理学、鼻腔灌洗液白三烯C4、外周血IL-4、鼻黏膜Toll样受体4和NF-κB蛋白表达等多个环节的影响,研究针刺疗法治疗过敏性鼻炎的机理,从细胞、分子水平揭示针刺疗法治疗过敏性鼻炎的机制。
1.实验动物与分组：SPF级健康SD大鼠40只,雌雄各半,体重180-220g。随机分为4组,即模型组、针刺组、西药组、空白组,每组10只；
2.实验动物造模：以卵蛋白注射及其鼻粘膜刺激法对针刺组、西药组、模型组建立变应性鼻炎动物模型；
3.干预措施：针刺组选取大椎、肺俞(双)、肾俞(双)穴进行针刺治疗,西药组以辅舒良气雾剂喷鼻治疗,模型组给予生理盐水滴鼻刺激,空白组不做处理；
4.观察不同处理组别卵蛋白所致过敏性鼻炎大鼠模型行为学及病理组织学改变；
5.观察不同处理组别卵蛋白所致过敏性鼻炎大鼠模型鼻腔灌洗液中白三烯的改变；
6.观察不同处理组别卵蛋白所致过敏性鼻炎大鼠模型大鼠外周血IL-4的改变；
7.观察不同处理组别卵蛋白所致过敏性鼻炎大鼠模型大鼠鼻黏膜TOll样受体4和NF-κB蛋白表达的改变；
8.分析各相关指标的改变意义,阐明针刺治疗过敏性鼻炎的免疫学作用机制。
三、研究结果
1.与空白组比较,模型组大鼠鼻痒、喷嚏和流清鼻涕等行为学方面积分显著增多(p<0.05),经过针刺治疗和辅舒良气雾剂喷鼻治疗后行为学积分与模型组比较有明显降低(p<0.05)
2.与空白组比较,模型组大鼠鼻粘膜嗜酸性粒细胞计数明显增高(p<0.05),经过针刺治疗和辅舒良气雾剂喷鼻治疗后鼻粘膜嗜酸性粒细胞计数与模型组比较有明显降低(p<0.05)
3.与空白组比较,模型组大鼠鼻腔灌洗液中LTC4含量明显升高(p<0.05),经过针刺治疗和辅舒良气雾剂喷鼻治疗后大鼠鼻腔灌洗液中LTC4含量与模型组比较有明显降低(p<0.05),西药组优于针刺组。
4、与空白组比较,模型组大鼠血清IL-4含量明显升高(p<0.05),经过针刺治疗和辅舒良气雾剂喷鼻治疗后大鼠血清IL-4含量与模型组比较有明显降低(p<0.05)。
5.与空白组比较,模型组大鼠鼻黏膜Toll样受体4蛋白表达平均光密度和NF-κB蛋白表达平均光密度明显升高(p<0.05),经过针刺治疗和辅舒良气雾剂喷鼻治疗后大鼠鼻黏膜Toll样受体4蛋白表达平均光密度和NF-κB蛋白表达平均光密度与模型组比较有明显降低(p<0.05)。
四、结论
1.针刺激肾俞(双)、肺俞(双)、大椎对过敏性鼻炎大鼠鼻痒、喷嚏、清涕等症状的改善有明显效果,即对过敏性鼻炎有较好的治疗效果,这进一步证实了针灸通过调理肺肾、升提阳气治疗过敏性鼻炎的正确性和合理性。
2.针刺治疗过敏性鼻炎大鼠的机理
(1)针刺治疗在一定程度上可以抑制炎症细胞,特别是嗜酸性细胞在气道内的聚集和活化,抑制嗜酸性细胞的活力,起到治疗气道炎症的作用。
(2)针刺可以降低过敏性鼻炎鼻腔灌洗液中LTC4的含量、降低血清中IL-4的含量及鼻黏膜TLR4和NF-κB蛋白表达,提示针刺对过敏性鼻炎的治疗效果是通过作用于多个环节而达到其治疗效果,特别是在对Toll-NF-κB信号通路上进行多层次调节。
Objective
Allergic rhinitis(AR) is allergenically inflammatory disease of type I induced by allergen, which is by The method of IgE and is characteristic by The esophocyte, mast cells, T lymphocyte. The symptom is nasal itch, sneeze and snivel with The complication of asthma. The abroad study revealed the occur of allergenic nasatitis is closely related to economic development. Recent years, the mobidity have inceased significantly to 10% with serious damage to the health of PeoPle.
AR is treated by modern medicine with immunal Therapy or durg, which have a lot of shortcoming. For immunal Therapy, The allergenic antigen is difficult to b3:determined, The effect is slow to occur, The course is too long to finish for the patients, The disease is difficult cured. The drug treating Allergenic nasatitis consist of durg anti-histamine, cortisol, atropine, with rapid effect and more side effect.
AR is calle "biqiu", "qiuti" in Traditional Chinese Medicine. Acupuncture treats the AR with long time and good results. But the immunal mechanism is undermined. In our study, the immunal mechanism of Acupuncture treating the allergenic nasatitis in investigated, which Provide scientific (?)rmation for therapy. Methods
1. Published Articles are Studied
The Published artieles are sudtied to declare the clinical and experimental investigation and provide the prospection.
2. Experimental Study
(?) our study, the effects of Acupuncture on The AR are investigation in (?)vation, pathology, TLC4 in NLF, IL-4 in blood, TLR4 and NF-κB in Nose mucosal.
2.140 SD rats, SPF and 180-220g weight are randomly divided into four groups:normal control group, model group, acupuncture treatment group and Fluticasone Propionate Nasal Spray(FPNS) treatment group.
2.2 Rat model of allergic rhinitis was prepared by injecting egg album and stimulating nasal mucosa.
2.3 Normal control group was not treated. acupuncture treatment group was treated by acupuncture on DU 14(Dazhui), BL 23(Shenshu) and BL 13(Feishu). FPNS group was treated by FPNS, model group was treated by physiological saline.
2.4 The behaviors of rats in all groups were observed. The pathological changes of nasal mucosa were tested and the contents of TLC4 in NLF, IL-4 in blood, TLR4 and NF-κB in Nose mucosal were determined by radio-immunologic analysis.
Results
1. Compared with normal control group, the behaviors'scores of model group were significantly higher (p<0.05). After treatment, the behaviors' scores of acupuncture treatment group and FPNS group were significantly lower than those of model group (p<0.05)
2. Compared with normal control group, the eosinophilic granulocyte(EOS) count of model group is significantly higher (p<0.05). After treatment, the EOS counts of acupuncture treatment group and FPNS group were significantly lower than those of model group (p<0.05)
3. Compared with normal control group, the serum content of TLC4 in NLF and IL-4 in blood of model group is significantly higher (p<0.05). After treatment, the serum content of TLC4 in NLF and IL-4 in blood of acupuncture treatment group and FPNS group were significantly lower than those of model group (p<0.05)
4. Compared with normal control group, the expression of TLR4 and NF-κB of model group is significantly higher (p<0.05). After treatment, the expression of TLR4 and NF-κB of acupuncture treatment group and FPNS group were significantly lower than those of model group (p<0.05)
Conclusion
1. Acupuncture treatment can effectively improve the symptoms of rats with allergic rhinitis. It means the method is effective for treating allergic rhinitis.
2. The possible action mechanisms of Acupuncture treatment in treating allergic rhinitis may be as the follows::①It can significantly decrease the EOS count of nasal mucosa of rats with allergic rhinitis.②It can effectively decrease the serum content of TLC4 in NLF and IL-4 in blood of rats with allergic rhinitis.③It can effectively decrease the expression of TLR4 and NF-κB of rats with allergic rhinitis.
引文
[1]王德鉴主编.中医耳鼻咽喉口腔科学[M].北京.人民卫生出版社,1994,第1版347357.
[2]王士贞主编.中医耳鼻咽喉口腔科学[M].北京,中国中医药出版社,2003,第1版,124-127.
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