Study On Pharmacodynamics And Mechanism Of Action For Asthma Rats From Maxinganshi Decoction In Different Ratio

As a famous prescription of Zhang Zhongjing's book Treatise on Cold-induced Febrile Diseases in Han dynasty,Ma Xing Gan Shi Decoction is used to clear heat and purge the lung,relieving cough and asthma.In this decoction,The property of Ephedra is acrid and disperse,which is used to disperse the lung and relieve asthma;Gypsum consumes more than doubled ephedra,so the acrid and warm property of ephedra weakened,and the entire efficacy turned into cold-pungent diaphoresis;almond is uesd to adjust lung-Qi,relieve asthma when cooperated with Ephedra;Liquorice is to modulate other herbs,since the compatibility of this decoction is ideal and easy to put it into clinical practice,it is widely used in cough,asthma and other symptoms by later physicians,especially applicable to evil heat in the lung. The prescription ratio between ephedra and gypsum and the role of gypsum and ephedra has always been a hot argument in physicians of past dynasties,different ratio of ephedra to gypsum(1:2-1:5)causing diffent efficacy in clearing lung heat and relieving exterior syndrome.The purpose of this study was to survey the pharmacodynamics and the mechanism of action in asthma animals by using different ratio of Ma Gan Shi Xing decoction,exploring the efficacy machanism in diverse proportion of ephedra and gypsum.bronchial asthma,a common respiratory tract disease as well as a chronic airway inflammatory result from eosinophilic granulocyte,mastocyte,T lymphocyte and other inflammatory cells involved in it's onset,severely threat public health in the worldwide.The morbidity of asthma is 2%in our country,but this rate will be up to 3%in the child.There are at least 15 million asthma patients in china according to the speculation.nearly 250 million people are suffering from asthma around the world.The statistic incidence rates varied by country,about 1 percent to 5 percent,of which children and women occupy an important proportion.With the industrialization process and the worsening environmental pollution,the data afforded by many countries indicates an upward trend of the incidence and the probability of asthma complications,but most cases falls into protracted and causes great financial burden as the poor clinical control and cure rate.asthma has become a serious social problems arousing the world attention.Therefore,the pathogenesis as well as the prevention and treatment research has been a hot research spot around the countries,the main purpose of asthma treatment is to inhibit airway inflammation and control symptoms at present.Through nearly 10 years of cytokines research progress in the pathogenesis of asthma,we found that a variety of cytokines involved in the asthma process of immune modulation (including the interleukin).The most important immune disorders is Th1/Th2 imbalance,Th2 cells and it's cytokines played a key role in induced asthma with typical pathophysiology characteristics.The role of T-lymphocytes in the bronchial asthma arouses increasing attention.in 1989,Mosmann first proposed Th2 type of hyperfunction and Th1 cell's hypofunction in the airway inflammation,Th2 type cells produce cytokines(IL-4,IL-5,IL-13)increase,while the cytokines(IL-2 and IFN-γ) produced by Th1 type cells declined,then result in the imbalance of Th1/Th2, that means the enhancement function of a subgroup,and weakened function of the other subsets,this phenomenon is known as"transfer cloning",which was also known as"Th1 and Th2 drift." There is a antagonistic relationship between Th1/Th2 cytokines.Nowadays the Th1 and Th2 cells are discriminated and determinated mainly through differentiating the IFN-γand IL-4 to reflect their level and function in the human body.Further research showed that,IL-4 plays a key role in causing asthma eosinophilic airway Inflammation since IL-4 can increase the quantity of immunoglobulin E(IgE),induce mastocytes divisived,and influence the growth,activation and chemotaxis of eosinophilic granulocytes.Then a lot of relevant studies have shown that a high Th2 type cytokines can be detected in the asthma pstients' peripheral blood,BALF (bronchoalveolar lavage fluid),and even the exhaled gas.gene knockout mouse model also prove that IL-4 and IL-13 can act on the airway,leading to it's hyperactivity. These cytokines research not only afford a high-specific target in the asthma treatment,but also provide the hope to control and reverse asthma-related inflammation.With the establishment of airway inflammation theory, anti-inflammatory treatment has been widely applied in clinical practice and acquired good therapeutic effect.Inhaled steroid has obvious anti-inflammatory effect,but can't reverse the natural course of asthma fundamentally,can't correct the potential immune disorder,furthermore, long-term useing may cause systemic side effects,while Traditional Chinese medicine has attracted more and more scholars' interest in a wide range,Ma Xing Gan Shi Decoction has been verified as an effective decoction for asthma treatment by the doctors of the past.This study aims at examining different ratio Maxing Gan shi decoction on asthma-like rats Th1/Th2 cytokines secretion of intervention,so as to penetrate the treatment of asthma in Chinese medicine into the mechanism of gene /priotein regulation and control,providing a scientific connotation for the therapeutic effectiveness of asthma in Chinese medicine.一,Pharmacodynamic Study for Ma xinganshi decoction in different ratioThis study showed that the ratio of different Maxinganshi decoction not only has an obvious effect on antipyretic,anti-inflammatory,relieving cough, but also has the significant inhibition for pneumococci,Staphylococcus,as well as influenza virus,parainfluenza virus infection,this result conform to Zhang Zhonghai's research using of rabbit fever model,the experimental research provide a scientific basis for the clinic application of maxinganshi decoction in different types of Exogenous fever,chest distress,coughing and other symptoms,the 1:5 high dose group's antipyretic,anti-inflammatory effect was better than 1:3 high dose group in the first and second hour,and The Ephedra (2)high-dose group showed the best therapeutic effect for the two nitro-fluorobenzene(DNFB)induced delayed hypersensitivity(DTH)in rats. For the impact on ammonia induced cough,there was no significant difference in different ratio group,so as for the antibacterial,antiviral activity of different ratio between the high-dose group and low dose group.二,mechanism of action for Maxinganshi decoction in different ratio1,effect on asthma rats high airway inflammation between different ratio decoctionThe result showed that there was significant difference between asthma model group BALF and control group(P＜0.05)for eosinophil EOS,monocytes and lymphocytes in the WBC count,indicating the successful model of asthma. There was an obvious increase in the number of EOS(P＜0.01)in asthma model group,while the EOS was significantly reduced between dexamethasone group, different Ma xinshigan decoction groups and asthma model group,The difference was significant(P＜0.05,P＜0.01),The dose-effect relationship exist in Maxing Gan Shi decoction for reduced number of EOS in asthma rats,especially for Ephedra(2)high-dose group,the therapeutic effect was similar to dexamethasone.As far as the pathological morphology changes in lung tissue at 400 times light microscope was concerned,an increase in bronchia duplicature,thicken alveolar septum and airway smooth muscle,the wall and alveolar interstitial was infiltrated by A large number of eosinophil, stenotic lumen could be observed in the asthma model group.While the inflammatory reaction was alleviated and the structure of mucous membrane was integral in the Hormone therapy group and different ratio Maxinshigan decoction group,the result indicated that this decoction has anti-inflammatory effect and certain dose-effect relationship,This may be related to the dosage of gypsum,the greater the amount of gypsum,the more obvious anti-inflammatory effect.2,The Impact on asthma rats Airway hyperactivity in different ratio Maxinshigan decoctionThis study was carried out by the method of egg albumin allergic so as to copy asthma rats model,testing local pathological changes in lung tissue and the infiltration condition of inflammatory cells in peripheral blood,and measure the sensitized animals airway hyperresponsiveness by the lung perfusion experiment,detect The airway hyperreactivity in vitro organ by spiral thread of trachead experiment,the result showed that the treated group has a decreased lung overflow when compared with the asthma model group (P＜0.01),while the difference was insignificant compared with control group(P＞0.05).The tracheal smooth muscle's contraction response to acetylcholine decreased significantly in different ratio Maxing Gan Shidecoction and dexamethasone group when compared with asthma model group,and the therapeutic effect in high dose decoction group was better than low dose group.There was no significant difference between Ephedra(1,2)high-dose group and control group(P＞0.05).this result indicated that this decoction could relax bronchial tension,thus reducing airway hyperresponsiveness,furthermore dose-effect relationship exists in different ratio group,the result was conformed to related clinical reports.Maxin ganshi decoction has repeatedly used ever since ancient times,mainly because its obvious effect in clearing heat and purging the lung,relieving cough and asthma.Clinical practice has proved that only by using several times or even 10 times the amount of gypsum to ephedra,can really experience it's ideal therapeutic effect.3,The effect of different ratio decoction on Th1/Th2 cytokines secretion①The effect on BALF,IL-4,IFN-γlevels and IL-4/IFN-γin the peripheral bloodThe result showed that compared with the asthma model,the BALF,total number of cells and EOS in the blood reduced in different ratio MaXin ShiGan group,BALF and IL-4 levels in the peripheral blood droped(P＜0.05,P＜0.01),And the effect was similar to dexamethasone group(P＞0.05), IL-4/IFN-γratio has correspondingly reduced,while IFN-γlevels increased,this means the decoction could inhibit Th2 cells subsets Preponderant response and regulate immune balance,the therapeutic effect was better in the high dose group than the low dose group,and there was insignificant difference between Ephedra(1,2)high-dose group and control group(P＞0.05)②effect on IFN-γmRNA and IL-4 mRNA in the lung tissueRT-PCR results showed the IFN-γmRNA expression level in Maxing Gan Shi decoction group was much higher than dexamethasone group(P＜0.05),the mechanism mainly through the adjustment of Th1/Th2,so that Th1/Th2 drift to Th1 and inhibit Th2 cytokines,then achieve the purpose of the treatment of asthma,the IL-4 mRNA expression level declined dramaticaly in the Dexamethasone group compared with the normal group and asthma group(P＜0.05,P＜0.01),while the IFN-γmRNA expression level showed insignificant difference compared with the normal group(P＞0.05),the difference was significant when coppared with maxinganshi group(P＜0.01),this means dexamethasone could inhibit IL-4 mRNA expression but not increase the IFN-γmRNA expression level simultaneously.Therefore,the conclusion could be drawed that maxinganshi decoction mainly increase the IFN-γmRNA expression and decrease IL-4 mRNA to correct the imbalance between Th1/Th2,while dexamethasone mainly restrain IL-4 mRNA expression but not increase IFN-γmRNA,In terms of immunoregulation,The former was a high-level bidirectional adjustment,and the latter was a low-level single directional one.③The effect on IL-4 and IFN-γprotein level in the lung tissueThere was an obvious low IL-4 protein level,high IFN-γprotein level,Lower eosinophilic granulocyte infiltration in the maxinganshi group compared with the model group.Therefore,We thought Maxin Shi-Gan decoction may affect the growth,activation and chemotactic process Of eosinophilic granulocyte.The mechanism may be mainly through the reduction of IL-4 protein leveland increase IFN-γprotein level.This decoction had more overall advantage since it doesn't weaken the body's immunity compared with the dexamethasone group.Conclutions:Different ratio MaxingGanshi decoction exerted a positive influence on the IL-4,IFN-γprotein levels and an obvious inhibited effect for the eosinophilic granulocyte infiltration in asthma rats.this maybe a good corrective action for the Th1/Th2 balance,thus have an impact upon curing asthma...