Comparison Of Inflammation Regulation Between Different Frequency Of Acupuncture Treatments In Mice Model Of Airway Hperresponsiveness

BackgroundAirway inflammation and airway hyperresponsiveness(AHR) are hallmarks and distinguishing features of bronchial asthma. In the process of researching the pathophysiology of the inflammatory response and the development of AHR, ethical and practical reasons frequently necessitate modeling asppropriate animal models.In most models of hyperresponsiveness,animals receive sensi-tization and challenge. The two challenge methods involve intranasal adminis-tration and nebulization. However, comparison between both of them are seldom investigated.As an incurable disease,most of patients in asthma take br-onchodilators and even glucocorticoids. But because of their side and untoward effects shown in public, the population of receiving acupuncture treatment with fewer side effects and lower price is ever-increasing, A large number of clinical research and animal experiments have proved that through regul-ating physical immune function acupuncture attenuate airway inflammatory responses.However, it is still a puzzle whether frequency of Acupuncture Treatments affects the effectiveness of inflammation regulation in mouse model of airway hperresponsiveness.Experiment 1 Comparison of the effects of different challenge methods on a mice model of hyperresponsivenessObjectiveTo investigate the effects of different challenge methods, through com-paring symptoms after 24h from last challenge.enhanced pause (Penh) after an inhaled stimulus(methacholine, Mch), structure of lung and the total number of alveolar lavage fluid leukocyte and then choose superior challenge method between them for experiment2.Methods26 female,6 to 8 weeks old, specific pathogen free Balb/c mice were divided into 3 groups including control group, intranasal challenge group and aerosol challenge group with 8 to 10 mice each group. Mouse in control group received intraperitoneal injection of 0.2ml/mouse sterile saline in day4,11,18. At the same days, the others were sensitized with a intraperitoneal injection of 50ug/mice chicken egg ovalbumin(OVA) emulsified in 2mg aluminum hydroxide adjuvant. On day 25-27, mice in aerosol challenge group were put into a plastic box with aerosolizedl% OVA for 30min once a day. At the same time,mice in intranasal challenge group received intranasal administration with 50ul1%OVA after anesthesia. To investigate the effects of different challenge methods, through comparing symptoms after 24h from last challenge, enhanced pause (Penh) after an inhaled stimulus (methacholine, Mch), structure of lung and the total number of alveolar lavage fluid leukocyte.Results(1)Symptom after challenge:in the process of nebulization, mice scratched nose more frequently and abdominal breathing occurred. After intranasal administration, tails and lips empurple.dyspnea occurred and 2 mice died in intranasal administration group. (2)Penh measurement:compared to control group, airway hyperreactivity of the others as evaluated increased signi-ficantly. But in low concentration of Mch, airway hyperreactivity had no statistical differences between aerosol challenge group and control group. (3) Total number of alveolar lavage fluid leukocyte when compared with control group, both of the other two groups as evaluated were larger with statistical differences.respectively. The same differences occurred in the latter two groups.Conclusionmost of test results showed that intranasal challenge group was superior to aerosol challenge group, but at the beginning of modeling with intranasal challenge method researchers were suggested to increase sample sizes appropriately in consideration of death rate.Experiment 2Comparison of Inflammation Regulation between Different Frequency of Acupuncture Treatments in Mice Model of Airway Hperresponsiveness.ObjectiveTo investigate whether the effects of different frequency of acupuncture treatments in mice model of airway hperresponsiveness were different and which was better.Methods38 female,6 to 8 weeks old, specific pathogen free Balb/c mice were divided into 4 groups including control group, model group,3 times acupuncture group and 7 times acupuncture group with 8 to 10 mice each group. Mouse in the latter three groups were sensitized and challenged as the same method as the intr-anasal group in experiment 1 above. The control group were injected and inhaled the same volume of saline instead. From day13, limbs of every mice were tied to a plank for 20 min/time once every 2 days by bookbinding cords. Such bind was repeated 6 times. During the period,7 times acupuncture group acupunctured Dazhui and Zusanli points for 7 times. But 3 time acupuncture group just performed the former 3 acupunctures. By comparing enhanced pause (Penh) after an inhaled stimulus (methacholine,Mch), structure of lung and the total number of alveolar lavage fluid leukocyte, the concentration of IFN-y.. IL-4 in BALF and IgE in plasma measured by sandwich ELISA.Results(1)Penh measurement compared to control group, airway hyperreactivity of the others as evaluated increased significantly. But in low concentration of Mch, airway hyperreactivity had no statistical differences between two acup-uncture groups. (2)Histologic studies showed that acupuncture dramatically inhibited lung tissue eosinophil infiltration and airway mucus production, especially in 7 times acupuncture group. (3) Total number of alveolar lavage fluid leukocyte when compared with control group, the other three groups as evaluated were larger with statistical differences, respectively. But statis-tical differences did not exist when the two acupuncture groups were compared to each other. (4) Compared to the control groups the concentration of IL-4 in BALF and IgE in plasma measured by sandwich ELISA in other groups there were statistical differences respectively, but statistical differences only existed between control group and model group when they compared concentration of INF-y in BALF to each other.ConclusionAs a whole considering the effect of reducing inflammatory response 7 times acupuncture group worked better than 3 times acupuncture group although both of them relieved inflammatory response effectively.