Trial Study On The Relationship Between Damp-heat, Damp-cold Enviroment And Expression Of Autoantibodies Related With RA

The pathogeny and the pathological of rheumatoid arthritis have not been known well and no specific medicines for it are found,so early diagnosis and proper treatment in the initial stage of the disease are important to prevent deterioration and disfunction of joints.Recent years along with the autoantibodies of RA are found it becomes possible to diagnose RA at it's initial stage.Quantity of researches indicated that there are some relationship between autoantibodies and severity of disease.Usually more autoantibodies are found more serious the disease is,and more earlier destruction of joints appears.Instructed by professor HuYinqi, We immunized mice with mixture of Freund's complete adjuvant(FCA) and CCP,two groups of immunized mice were put into damp-heat and damp-cold environments respectively to research the effection of environment to the AA,and the effection to RA autoantibodies;The relationships among environment,autoantibodies and the expression of IL-1 and VEGF in gristle cells were also been studied in order to seek a method to make disease-syndrome combined animal model of RA. 1 Literature studiesAKA,APF,anti-Sa,anti-CCP can be detected in initial stage of RA,and all these autoantibodies are specific for RA,though they are not so sensitive to it.Studies show that they also related with destruction of joints of RA.That three or more than three antibodies are detected is aimportant marker of danger of destruction joints.IL-1 and VEGF abnormal excretion are the basic factors of important pathology of bone,cartilage erosion and synovial membrane over-growth.It can been judged which is the danger marker of joints erosion by research the relationship among IL-1, VEGF and autoantibodies expression and environment of damp-cold and damp-heat.RA disease-syndrome animal model generally made by affixing environment factor according to syndrome character to the basic model of modern medicine of RA.On the basis of this principle ,authors have produced many disease-syndrome model of RA,for example :the model of wind-damp-cold model,damp-heat model,phlegm-blood stasis model and kidney deficiency model,et al.All of these models have some features of disease and syndrome in certain degree,but they have not detected autoantibodies which are so important for early diagnosis of RA,and play a great role in the pathologic process of RA. So it is possible to produce disease-syndrome model with autoantibodies of RA to immunize mice with mixture of FCA and CCP.2 Experimental study2.1 Methods of experiment2.1.1 Group dispensation32 healthy SD mice were divided into 4 groups (normal group,model group damp-cold group and damp-heat group) in random .Each group contains 8 mice.2.2.2 Model making process and methods of experiment CDModel group:8 mice were immunized with the mixture of 0.1ml FCA and 100 ~ 150jj.g CCP which was injected into under bottom skin of both feet. After two weeks, lml blood was taken from tail of every mouse to dispart serum for detection.A month late, 4 ~ 6ml blood was taken from heart of every mouse and dispart serum for detection.?Damp-heat group: 8 mice were treated as the model group except that they were put into damp-heat environment one hour a day for nine days which was 30 ~ 33 °C of temperature and 95% ~ 100% of humidity.?Damp-cold goup: 8 mice were treated as the model group except that they were put into damp-cold environment one hour a day for nine days which was 7 ~ 10°C of temperature and 95% ~ 100% of humidity.?Normal group: No treatment was given, after two weeks ,1ml blood was taken from tail of every mouse to dispart serum for detection.A month late, 4 ~ 6ml blood was taken from heart of every mouse and dispart serum for detection.?At the end of experiment, both ankle joints were cut off for pathological examination. 2.2.3 Items observed?General detection: mice weight,thickness of feet.?Autoantibodies detection: AKA,APF,anti-CCP,RF and anti-Sa.?Pathological detection:ankle joints pathological detection andVEGFJL-1 expression in cells of cartilage of ankle joints. 2.2.4 Trial results:(J) Mice weight of damp-cold and damp-heat groups increased slowly and there was a obvious discrepancy when compared with other two groups,the weight of damp-heat group mice increased more slowly than mice of damp-cold group.Both environments aggravated the swollen of mice feet,it seems that the damp-heat group had been seriously effected by environment,but there was no distinctive discrepancy when compared with damp-cold group.(Dig and CRP: The contents of IgA,IgM,IgG and CRP in serum of model group,damp-heat group and damp-cold group were much higher than normal group.At the end of two weeks, IgA,IgM of damp-heat group were much higher than other groups, while IgG of damp-cold group was much higher than other three groups.At the end of trial there was no significant difference between damp-heat and damp-cold groups for Ig in serum.At the end of two weeks and a month, the content of CRP in serum of damp-heat group was higher than that of damp-cold group and model group,and that of damp-cold group is higher than model group.(3)RF and anti-CCP:At the end of two weeks and a month, the contents of RF and anti-CCP in serum of damp-heat group,damp-cold group were much higher than that of model group,but there was no difference between damp-cold and damp-heat groups.(D RA33: At the end of trial the level of RA33 in serum of model group ,damp-heat and damp-cold group are higher than normalagroup,but there is no significant discrepancy among these three groups.?AKA and APF: At the end of two weeks and a month, the contents of APF and AKA in serum of model group,damp-cold group and damp-heat group are much higher than that of normal group;At the end of two weeks, there were no significant difference among model group,damp-cold group and damp-heat group ;At the end of a month AKA and APF were much higher expressed in serum of damp-heat group when compared with other three groups.?IL-1 and VEGF: At the end of trial, the mean area of IL-1 and VEGF positive cartilage cells of damp-heat,damp-cold and model group was much larger than normal group,there was a significant difference between them,while there were no significant difference among these three groups of model group,damp-cold group and damp heat group.There were more IL-1 and VEGF positive cartilage cells in joints of damp-heat,damp-cold and model group than normal group,and there were more VEGF positive cartilage cells in damp-heat group than other three groups,while there were more VEGF positive cartilage cells in damp-cold group than other two groups.?Pathological results .-Normal group :The structure of bone and cartilage of joins was intact without infiltration of inflammatory cells in synovial membrane and synovial membrane over-growth;Model group:The bone and cartilage structure of joints was almost intact without distinctive synovial cells over grown;Damp-cold group:Synovial membrane of joints over grownwith infiltration of inflammatory cells and 3 ~ 4 lays of synovial cells aligned in disorder.Bone beams getting sparse with enlarged medulla cavity;Damp-heat group: Synovial membrane of joints over grown with infiltration of inflammatory cells and several lays of synovial cells aligned in disorder. Sparse bone beams cracked and laid disorderly . 2.4.5 Conclusion:?Environment factor made AA model characterized with syndrome feature of TCM in certain degree, so it can been used as a method of making disease-syndrome animal model of TCM. (D Enviroment factor aggravated immunity react ion,synovial cells over growth, osteoprosis and bone beams crack of animal model,madeIg, RF, CRP, anti-CCP,APF,AKA ,VEGF and IL-1 in positive cells expressed much stronger than models without environment factor.?There were more IgA, IgM, RF, CRP, anti-CCP, APF, AKA, RA33 in serum of animal model which in damp-heat environment, VEGF and IL-1 are expressed much stronger than that in damp-cold environment; The pathologic influence of damp-heat environment was more serous than that of damp-cold environment.?There was no significant discrepany for influence of RF, anti-CCP and VEGF, IL-1 positive cells between damp-cold and damp-heat environment.