Dehydroepiandrosterone (DHEA) Affect The Level Of IFN-γ,IL-4,IgE In Serum Of The Asthmatic SD Rat Models

Background and objectives: Bronchial asthma (abbreviating asthma) is a chronic airway disease characterized by chronic inflammation.The level of immunoglobulin E(IgE) and the number of EOS increase in the body of patients with asthma.There mainly are Eosinophils(EOS),Lymphocytes,Neutrophils,Basophils and so on in the lavaged solution from the asthmatic pulmonary alveolar.The principle is complicated which is inflammatory cells getting together into the airway.Firstly,After IgE insert into the FC receptor of the mast cells, the mast cells allergized.when the mast cells retouch antigent, the mast cells will release its granular,which hold many material such as histamine ,leukotriene,cationic protein,platelet activating factor ,and so on.Then those materials result in airway inflammation and bronchi spasm.Secondly,On the contrary,The inflammatory cells also secrete cytokine to aggravate airway infammation,which include interleukin 4(IL-4), interleukin 5(IL-5), interleukin 2(IL-2), interleukin 13(IL-13), interleukin 8(IL-8) and so on.Now,it is mainly thought that IL-4 contribute to secreting the quantity of IgE .The IL-5 enlarge the function of IL-4 in secreting IgE. To think at present,the function disorder of help CD₄⁺T(TH) cells is a reason to cause IL-4 risingAccording to help CD₄⁺T cells secreting different cytokine , CD₄⁺T cells are divided into two subgroup, TH₁ and TH₂. THi cells mainly secrete cytokine such as interleukin 2(IL-2), interleukin γ (IFN-γ),tumor necrosis factor β (TNF_β) and so on,while TH₂ cells mainly secrete cytokine as follow IL-4, IL-5, IL-6 and so on.In the body of patients with asthma,the effect of TH₂ cells secreting cytokine rise,while the effect of TH₁ cells secreting cytokine decrease,so that the cytokine of IL-4 and so on increase to resultin high level of IgE. THi and TH2 cells are differentiated from THo cells.There are many factor to affect the process of differentiating,including purified protein derivative of tuberculin ,antigen-presenting cells(APC),local microenvir- onment, cortex Hormone and so on.Among cortex hormones, Dehydroepiandrosterone is a kind of hormones,which can promote TH0 cell to differentiate into THi cell and enlarge the function of THi in secreting cytokine.DHEA is the most abundant steroid material in the human blood circulation,and exist in the blood circulation by its sulfate ester.The chemical name of DHEA is 3 P -hydroxide radical androst-5-alkene-17-ketone,which is produced by the adrenal cortex through cytochrome p450 main enzyme catalyzing cholesterol to synthesize.In the fetus, DHEA is produced by the placenta through the sulfate ester enzyme of the placenta catalyzing cholesterol to synthesize.In the brain,the oligodendrocyte synthesize DHEA.During producing,the composite process is not affected by the endocrine material all through body.Dong zong qi consider that the key mechanism of the asthma disease occurring include such as the ratio of THi/ TH2 unbalance, IFN- Y reducing,a cytokine which THi cell secrete, IL-4,IL-5, IL-6 increasing ,the cytokine which TH2 cell secrete.lt is promising to find new therapy to cure of the patients with asthma by modulating the disorder of THi/ TH2 cells. It is discovered by researching in molecule level that the mechanism of specificity immune therapy is to modulate the disorder of THi/ TH2 cells about immune response.The proof of attaining normal level of THi/ TH2 is that IL-4 by far reduce ,so that IgE also reduce ,while IL-2, IFN-Y by far rise.In the body of the patients with systemic lupus erythematosus,some researchers have observed that DHEA can strengthen CD4+T cells activated by phytohemagglutinin(PHA) to secrete IL-2 and IFN- Y which is the most evident in the physiological concentration of DHEA .At present,It is not reported that how DHEA and THicells affect the asthma ,so ,to explore new method to cure of asthma ,the writer will study that DHEA enhance the function of THicells secreting cytokine.The objective of the experiment by making the asthmatic SD rat models is toinvestigate the level of IL-4, IFN- y ,IgE in serum of the asthmatic SD rat models that is treated by DHEA ago and after.And we compare the level of IL-4, IFN- Y ,IgE in serum of model rats with the level in control group .The concentration of IL-4, IFN- Y ,IgE in serum of the asthmatic SD rat models is detected with the method of Enzyme-linked immunosorbent assay(ELISA).Materials and methods rExperiment animals:19 female and male SDrats,clear grade,weight 150-250g.Drug and Agent: DHEA, which is donated by Yong zhou medical factory.Ovalbumin(OVA) ,which is the production of Sigma company in America and is bought from the biology technique limited company Beijing.The agent box of Enzyme-linked immunosorbent assay of IL-4, IFN-Y ,IgE,which is gotten from Wu han Boster Biological Technology limited company. 1> 2-propanedioL Ethanol absolute ^ aqua proinjection ,which agent is analysis pure agent in market and is bought from the market.The each of 30 female and male SD rats is injected with lml 10% OVA solution containing OVA 10mg,aluminium hydroxide, 100mg,and aqua proinjection into their abdominal cavity,on 1st and 14th day.Total times is two.From 21th day beginning,30 asthmatic model rats injected with 10% OVA solution are separated into several bulk to inhale 1 % OVA solution through ultrasound atomization machine in the manmade organic glass box that is close(30x25x20cm) so as to agitate asthma.Each time of the atomization last 15-30 minutes,a time a day,total 5 days.Then 30 the asthmatic SD rat models are randomly divided into two groups.15 models for interfering group,which is injected with DHEA,5mg DHEA into their abdominal cavity each rat every time . DHEA is dissolved into solution by 5:4:1 of U 2-propanedioh Ethanol absolute > aqua proinjection. lml the solution contain 5mg DHEA.Each of 15model rats is injected with lml the solution into their abdominal cavity,on 28th and 35th day.Another group for control group,15model rats,which is injected with lml the solution of 5:4:1 of 1> 2-propanediol> Ethanol absolute^ aqua proinjection into their abdominal cavity,each rat every time ,on 28th and 35th day.Total injecting time is two.The blood of 15 interfering group model rats are collected from their tails on 27th and 42th day,eachrat 1.5ml blood. 15 model rats for control group are collected their blood from their tails on27th and 42th day ,each rat 1.5ml blood.Then the 20ul blood is gotten out from each 1.5ml blood to count the number of EOS,the other blood specimen is separated to get the serum .Preserve 150ul serum specimen into -70 °C refrigerator to wait testing.According accurately to the method and step of the instruction of ELISA,we detect the serum specimen.At last,we add stop solution to each detected hole within 30 minutes.The OD number of the detected serum is gotten with the Enzyme-linked immunosorbent assay Metertech.The concentration of the material to test is gotten with the Enzyme-linked immunosorbent assay Metertech,too.Every data is shown in mean and standard( x ±s).The comparison of each group of data is to use K-W rank test or two-sample t-test for independentsamples.P value less than 0.05 indicate statistical significance.Results:The concentration of IFN- Yof the serum (84.41±31.37pg/ml)in the model rats of the interfering group after injecting DHEA is by far more high than control group (57.87+8.92pg/ml), H=10.981, P<0.004.The concentration of IFN- Y of the serum in interfering group after injecting DHEA is more high than the interfering group before injecting DHEA (65.99±12.23pg/ml),too. H=4.477,P<0.034.While the concentration of IFN- Y of the serum in the interfering group before injecting DHEA is like the control group , H=2.2.687, P>0.101.1. The concentration of IL-4 (2.47±3.39pg/ml)in serum in interfering group after injecting DHEA is by far lower than control group (7.70±22.36pg/ml). H=12.265, P<0.0001.The concentration of IL-4 in serum in interfering group after injecting DHEA is by far lower than the interfering group before injecting DHEA (10.59±4.84pg/ml) too. H=18.835, P<0.0001.While the concentration of IL-4 in serum in control group is like the interfering group before injecting DHEA. H=1.603, P>0.205.2. The concentration of IgE in the serum (1.2667±3.34pg/ml)in interfering group after injecting DHEA is by far lower than control group (29.378±28.49pg/ml). t=-3.794,P<0.001.The concentration of IgE in serum in interfering group after injecting DHEA is lower than the interfering group before injecting DHEA (23.293±21.54pg/ml) too. t=-3.914,P<0.001.While the concentration of IgE in the serum in the interfering group before injecting DHEA is same like control group . t=0.66,P>0.515.Conclusion:1. DHEA increase by far the concentration of IFN- Y in serum of the asthmatic SD rat models.2. DHEA decrease by far the concentration of IL-4 in serum of the asthmatic SD rat models.3. DHEA decrease by far the concentration of IgE in serum of the asthmatic SD rat models.4. DHEA affect the immunity of the body in the asthmatic SD model rats so that the disorder of THiATH2 in the asthmatic SD models rats get normal.