Protection And Mechanism Research Of Yiqiyangyin Decoction To Leucocyte Of Experimental GD Mice

ObjectiveThe research is based on traditional chinese medicine to lunch a protection and mechanism research of Yiqiyangyin decoction to leucocyte of experimental GD mice.we used gene gun to establish mice model and then detected peripheral white blood cell count of mice,thyroid function(FTI),pathological change of thyroid histology,apoptosis-related genes,IL-13, PPAR-γand other indexes.we gave a in-depth exploration of protection and mechanism research of Yiqiyangyin decoction to leucocyte of experimental GD mice,made a preliminary revelation of it,provide pharmacological basis,and accomplish its research; we can improve curative effect of Yiqiyangyin decoction to GD and its complications,reduce the toxic side effects of western anti-thyroid drugs to further improve Chinese medicine characteristics and advantages;meanwhile,we compared the efficacy of different doses of the prescription, probe into its prescription-making rationality, lay the theoretical foundation and experimental basis for further drug discovery.MethodsGD animal model established by female Balb/c mice immunized with gene gun were random into five groups:1 model group;2 C-H group;3 C-M group;4 C-L group;5 methimazole group;6 normal control group. All the group were given intervening therapy with high,middle,low dose of Yiqiyangyin decoction or western medicine, RIA was used to detect TH;immunohistochemistry to nuclear transcription factor PPAR-γ;RT-PCR to apoptosis-related genes expression of IL-13;FCM to marrow cell circle and expression of CD45 antigen.Results1 Experimental result of mice peripheral white blood cell countC-H group, C-M group, C-L group, model group and normal control group had no significant difference, P>0.05;but compared with western medicine, the difference is significant,P<0.05;there had significant difference between western medicine group and normal group with P<0.01;there has significant difference between model group and normal group with P< 0.05.2 Experiment result of GD mice THThe mice serum FT3 and FT4 of model group increased obviously, with a significant difference compared with normalgroup (P<0.01),especially FT4,which was shown that our model-making were successful.There had significant difference between C-M group and C-H group, P<0.05;There had very significant difference between C-H group with C-L group, P<0.01;which manifested that C-H group had a more effective result than C-M group and C-L group in improving thyroid function. C-H group, methimazole group and normal group had no significant difference,P>0.05;3 Experiment result of mice marrow cell CD45 molecule expression.CD45 molecule expression was showed by the track numbers CD45+ accrossed. The CD45+ cell of normalgroup has the highest through rate,with 99.59%:the through rate of CD45+cell of C-H group(98.57 %),C-M group (98.39%),C-L group(98.39%)had significant difference with model and normal group,P<0.01;methimazole group(94.22%)had significant difference with C-H group, C-M group and normal group,P<0.01.model group (91.26%) had significant difference with C-H group, C-M group,C-L group,P<0.01;but it had no significant difference with normal and methimazole group, P>0.05.They all showed that after the treatment of Yiqiyangyin chinese medicine,the CD45 molecule expression of mice marrow cell increased,which also mean the track number CD45+ cell passed increased.4 Experiment result of the expression of mice marrow cell apoptosis-related gene-bax,bcl-2mRNABaxmRNA expression ability of model group had modest reduction,which has no significant difference with blank group, P>0.05;the tapazole group increased apparently, which has significant difference with normal group,P<0.01;there have no significant difference among C-H group, C-M group,model and normal group,P>0.05; there have significant difference among C-L group,tapazole,modeland normal group, P<0.01.Bcl-2mRNA expression ability of model group had modest increase,which has no significant difference with normal group, P>0.05;the tapazole group decreased markedly,which has significant difference with normal group,P<0.01;there have no significant difference among C-H group,C-M group, model and normal group,P>0.05;C-L group, tapazole group have significant difference with model group, P<0.05;C-L group has significant difference with normal group, P<0.05;tapazole group has significant difference with normal group,P<0.01.5 Experiment result of mice IL-13mRNAIL-13mRNA expression ability of model group had modest decrease, which has no significant difference with normal group,P>0.05;the tapazole group decreased markedly,which has significant difference with normal group,P<0.01:there has no significant difference in the IL-13mRNA expression between chinese medicine treatment group and normal group,P>0.05. After the treatment of Yiqiyangyin chinese medicine,the expression of mice IL-13mRNA has made a down-regulation, which mean C-H group express the lowest,with gradually increased.these all show that the Yiqiyangyin chinese medicine made a promoter action in the ripe,activization,mutiplication of immune cell and immune regulation.6 Experiment result of mice PPAR-γThe PPAR-γimmunoreactivity cells were sediment whose nucleolus dyed claybank or had claybank particle.mice PPAR-γexpression level of model group was less,and after the treatment of Yiqiyangyin chinese medicine,its expression level turn to up-regulation. the mice PPAR-γexpression of C-H group increased markedly(++++),and C-M group (+++);gradually down-regulation.Conclusion1.GD animal model established by female Balb/c mice immunized with gene gun expressed GD animals main characteristic,and were feasible and applied in pharmic and pathological research2.Chinese medicine Yiqiyangyin decoction can improve TH level and cure GD patients.C-H and C-M are equivalent in treating GD patients with tapazole,however,C-L is much less.3. Not only can Chinese medicine Yiqiyangyin decoction treat GD patients,but also not lead to the reduction of eripheral white blood cell count, which avoid the virulent and side effect of western medicine. Its effects is superior than tapazole.C-H and C-M is better than C-L.4.The protection of TCM Yiqiyangyin decoction to GD white cell reflect in it can regulate Graves mices marrow cell apoptosis level,restrain marrow cell apoptosis and make it get to right level.The point of restraining marrow cell apoptosis,rising white cell,and improving immune system may be the mechanism of action for Yiqiyangyin decoction to protect white cell,and Bcl-2,bax gen may be its target.