The Effect Of Point Application And Acupoint Injectionof Utilins On Airway Inflammation And Hyperresponsiveness In OVA Induced Asthma Rats

Objective:1.To observe the effectiveness of acupoint application combined with acupoint injection of Utlins on airway inflammation and airway hyperresponsiveness.2.Through the inflammatory cells in BALF,IL-4 and IL-5 in lung tissue,IL-4 and IgE in serum,NF-? B enter into the cells nucleus.We study this treatment methods on airway inflammation.3.Through the monitoring of ET-1in lung tissue.We study the treatment methods on airway hyperresponsiveness.Methods:Sixty-five cleaning Wistar rats(male,4 weeks,weight90-110grams)were randomly divided into 5 groups,normal group,asthma group,the point application and acupoint injection group,the Budesonide atomization group,the combined curative group.13 rats in each group.The asthma rats were sensitized on the first day and the 8th day with 1ml 10%antigen liquid(contains 100 mg ovalbumin and 100 mg AL(OH)3).And then these rats were inhaled aerosolized OVA(2%)in a not completely closed atomization chamber from the 15 th day.The rats were inhaled,30 min every time,once a day,until the rats appears to be agitated,choking cough,shortness of breath,nodding breathing,slow moving,cyanosis in tail even though abdominal cramps.These symptoms can be observed appears to early stage.At this time we test eostinophils(EOS)is high in blood,observed the infiltration of inflammatory cells in lung tissue is obvious under the microscope.With all of these,the model is successful.The normal group was given normal saline to replace OVA,method as above.Treatment begins with the model is successful.The point application and acupoint injection group:pasted the Chinesemedicine on the dazhui point,double dingchuan point,double feishu point,double shenshu point;injected Utilins on the double dingchuan point/double feishu point alternately,three times a week.The Budesonide atomization group: aerosolized Budesonide every day.The combined curative group: point application,acupoint injection,aerosolized Budesonide,three kind of methods used in combination.A total of one month(12 times).Stimulate3 times in a week to simulate a chronic asthma duration.The rats were sacrificed within 24 hours after the last inhalation.The BALF,blood and lung tissue of the rats were collected.The number of total cells,neutrophils,lymphocytes and eosinophils in the BALF were detected.Part of the lung tissues were sliced and stained with HE.,HE staining was used to observe the morphological changes.The concentration of IgE and IL-4 in serum was measured through ELISA.The concentration of IL-4?IL-5 and ET-1 in lung tissue were measured through radioimmunoassay.We measured the expression of NF-? Bp65 in lung tissue by immunohistochemistry respectively,the relative expression of NF-? Bp65 was calculated by the computer image analysis software.Results:1.Compared with the normal group,The infiltration of inflammatory cells and airway injury are more serious in asthma group.Inflammatory cell score was higher.Compared with the asthma group,Inflammatory cell score was lower in the treatment groups.Both of the difference is statistically significant(P<0.05).2.Compared with the normal group,the weight was lower,the levels of total number of cells,percentage of neutrophils/ Lymphocytes/ Eosinophilsin BALF;IgE and IL-4 in serum;IL-4/IL-5/ET-1;the IOD of NF-? B positive cells were higher in the asthma group.All of the difference is statistically significant(P<0.01).3.Compared with the asthma group,the level of total number of cells,percentage of neutrophils/ Lymphocytes/ Eosinophils;IgE and IL-4 in serum;IL-4/IL-5/ET-1;the IOD of NF-?B positive cells were lower in thetreatment groups(all P<0.05).The weight in no Statistically differences(P>0.05).4.Comparison between the treatment groups: the level of total number of cells,percentage of neutrophils/ Lymphocytes/ Eosinophils;IgE and IL-4 in serum;IL-4/IL-5/ET-1;the IOD of NF-?B positive cells and ET-1in the lung tissue in the combined groups were lower than the others treatment group(all P < 0.05).Compared with the point application and acupoint injection group,the ET-1is no Statistically differences(P>0.05).5.Correlation analysis: There was a positive correlation between the IL-4/IL-5 in the lung tissue and the IOD of positive cells(r=0.952,P < 0.01)/(r=0.931,P<0.01);The same to IL-4 and IgE in the serum(r=0.854,P<0.01);the IL-5 in the lung tissue and the percentage of EOS in BALF(r=0.904,P<0.01).Conclusion:1.By the antigen sensitization and repeated nebulization,OVA-inducded asthma model of rats was successfully established.In asthma group of pathologic results,we can find that airway wall were injured and inflammatory cells infiltrated around the airway,smooth muscle were thickness.2.In the asthma group,the changes of inflammatory cells;IL-4?IgE in serum;ET-1?IL-4?IL-5?NF-?B in lung tissue had a relationship with the happens of asthma.These indicators could evaluate the effectiveness of the treatment.3.The effect of using acupoint and acupoint-injection was poor than budesonide,the combined effect is more obvious.The mechanism may be(1)To reduce the inflammatory cytokine of Th2 by inhibiting the activation of NF-?B(2)To reduce the hyperresponsiveness of the airway by inhibiting the level of ET-1.