| Obiective: To observe the effect of acupoint injection of mycobacterium and acupoint application of Chinese herbs on serum Imunoglobulin E(Ig E)level,the expression of Interleukin-10(IL-10),Interleukin-17(IL-17)and Intercellular adhesion molecule-1(ICAM-1)in lung tissues in asthmatic rats.To explore the mechanism of external treatment of traditional chinese medicine(TCM),as acupoint injection,acupoint application on bronchial asthma.To provide a strong basis and reference for the treatment of bronchial asthma by mechanism of external treatment of TCM.Methods: 60 SPF grade Wistar male rats were randomly divided into control group,model group,mycobacterium acupoint injection group(acupoint injection group),acupoint application of TCM group(acupoint application group),mycobacterium accupoint injection and acupoint application of TCM group(injection and application group),each group including 12 rats.Rat model of bronchial asthma in the preparation of ovalbumin(OVA),including the stages of sensitization phase and excitation phase.In addition to the control group,the remaining rats were injected with ovalbumin and aluminum hydroxide mixture 1ml(10% of OVA and 10% aluminium hydroxide),eighth days once again.The control group by intraperitoneal injection of the same amount of saline solution.15 days begins,the rats were placed in a airtight atomization box,to 2% OVA solution ultrasoinc atomization,the control group with saline atomization.Every second day,30 min.Rats were observed behavioral change.When there is an agitated,scratching nose,cough,breathing rate to speed up the rate increase,obvious abdominal breathing,nod to breathe,even limbs collapsed and incontinence,that asthma is triggered.Atomization 7 consecutive days,then 2 times per week.The control group of normal feeding and dose not do other processing.The rest of the group were treated by chosing the cupuncture Dazhui,Dingchuan,Feishu,Pishu,Shenshu.Acupoint injection group: five acupoints daily take turns to inject of mycobacterium.Acupoint application group: stick the TCM paste 2 outside the points,6 hours before removal.Injection and application group: stick after injection,6 hours before removal.Control group acupoint selection the same as the treatment group,instead with the same amount of saline solution and an empty post.Operate once every other day,three times for a period of treatment,a total of four courses.HE staining of lung tissue to observe the airway inflammation,ELISA method to detect serum IgE,and the content of IL-10,IL-17 in lung tissue homogenate;Immune histochemical method to detect the expression of ICAM-1 lung tissue.Results:1 Behavioural performance of rats in each group:The control group rats breathe smoothly,curled up when atomization,without shortness of breath,and other special performance;When the model group is atomized,the rats had a higher frequency of breathing,mouth breathing,nodding breathing,sneezing,scratching,irritability,and abdominal muscle spasm,nose and nails at cyanosis,even slow,limp,incontinence,etc.With the increase of the number of times the symptoms gradually increased,1 died of asthma attack.After several times of stimulation,rats diet significantly reduced,the hair color withered,vertical hair,etc.The rat model has the characteristics of asthma.The above symptoms of rats were not obvious in the acupoint injection group,acupoint application of TCM group,acupoint injection and application group.Visible irritability,nodding like breathing,scratching nose,but lip nail cyanosis reduced,individual visible lying motionless,fecal incontinence.However,with the reduction of the number of times and the increase in the number of treatment,the rats symptoms gradually improved,acupoint injection plus sticking group improved significantly.2 HE changes in lung tissue:Control group: The bronchial epithelium was complete,the shape was regular,and there was no hyperplasia of the bronchial tube wall,the regular rules of alveolar morphology,no rules of alveolar wall,hyperplasia and inflammatory infiltration.Model Group:Around the trachea and lung tissue inflammatory cell infiltration,bronchial stenosis deformation,spacesvisible exfoliated epithelial cells,mucosal fold increase and edema,alveolar wall thickening and pulmonary septum,congestion,alveolar cavity decreased or even disappeared.Mycobacterium acupoint injection group: Compared with the model group,the inflammatory cell infiltration was reduced,the bronchial morphology was slightly changed,the mucosa was slightly thickened,and the pulmonary alveolar wall and pulmonary septal thickening were alleviated.The alveolar in irregular form.Acupoint application of traditional chinese medicine group: The morphology of the bronchioles was more regular,there were no obvious epithelial cells shedding,inflammatory cell infiltration in the wall of the tube,thickening of the mucosa,hyperplasia of alveolar wall and lung,and irregular morphology of the alveoli.Acupoint injection combined with application group: The bronchioles had regular shape,there was a small amount of inflammatory cells in the periphery of the tube.The wall mild hyperplasia,the mucosa and alveolar cavity are almost normol.Individual alveolar wall thickening and inflammatory cells have increased.3 Determination of IgE content:The serum IgE of the model group was significantly higher than that of the control group,there was significant difference(P<0.01);The content of IgE in the three treatment groups was lower than that in the model group,the difference was statistically significant(P<0.01);The content of Ig E in the acupoint injection and application group was lower than that in the acupoint injection group and low in the acupoint application group(P<0.05);There was no significant difference in IgE content between acupoint injection group and acupoint application group(P>0.05).4 Determination of IL-10 content:The expression of IL-10 in lung tissue of model group was significantly lower than that of control group(P<0.01);The content of IL-10 in the three treatment groups was higher than that in the model group,the difference was statistically significant(P < 0.01);The content of IL-10 in the acupoint injection combined with application group was higher than that in the acupoint injection group and the acupoint application group(P<0.05);There was no significant difference in IL-10 content between acupoint injection group and acupoint application group(P>0.05).5 Determination of IL-17 content:The expression of IL-17 in lung tissue of model group was significantly higher than that of control group(P<0.01);Compared with the model group,the content of IL-17 in the three treatment group was decreased,and the difference was statistically significant(P<0.01);The content of IL-17 in the acupoint injection combined with application group was lower than that in the acupoint injection group and the acupoint application group(P<0.05).There was no significant difference in the content of IL-17 in the acupoint injection group and the acupoint application group(P>0.05).6 The expression of ICAM-1 was observed by immunohistochemistry:The control group showed a few positive cells;In the model group,there are more expression of brown granules in bronchi,bronchial secretions and interstitial lung,alveolar cavity;Around the acupoint injection group and the acupoint application group of bronchus,pulmonary interstitial alveolar,brown particles less than the model group;Around the acupoint injection application group,bronchus,pulmonary interstitial and alveolar,brown positive cells expression is not obvious.Comparison of average optical density in each group: the expression of ICAM-1 in lung tissue of model group was significantly higher than that of control group(P<0.01);Three the expression of ICAM-1 in the treatment group was lower than that in the model group,the difference was statistically significant(P<0.01);The ICAM-1 expression of in the acupoint injection combined with application group was lower than that in the acupoint injection group and the acupoint application group(P<0.05).There was no significant difference in ICAM-1 expression between acupoint injection group and acupoint application group(P>0.05).Conclusions:1 According to the symptoms of the model group and the detection indicators,HE staining showed that the rat model of bronchial asthma was successful.2 The content of IgE increased,the content of IL-10 decreased,the content of IL-17 increased,and the expression of ICAM-1 increased in lung tissue of asthmatic rats,The changes of asthma symptoms and lung pathological changes in rats were consistent.These results suggest that IL-10,IL-17,IgE,ICAM-1 are involved in the pathogenesis of asthma.3 Acupoint injection and acupoint application may be by reducing the level of IgE in asthmatic rats,enhance the expression of IL-10 in lung tissues of asthmatic rats,the decreased expression of IL-17,reduce the expression of ICAM-1,to reduce airway inflammation,regulating immune balance,comprehensive application of the two methods compared with the single application effect is better. |
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