| ObjectiveTo further study the therapeutic effects of eliminating goiter recipe and formular wenshenjianpi and formular sanjie-xiaoying by using them for intervening and curing model rats of autoimmune thyroiditis and to define its pathogenesis from the aspects of apoptosis,immunoreaction and cytokines,and to discuss the traditional Chinese medicincal pathogenesis of autoimmune thyroiditis from traditional Chinese medicincal view of phlegm-blood stasis and deficiency of spleen and kidney with the expectations of deepening the knowledge of the pathogenesis and traditional Chinese medical pathogenesis of autoimmune thyroiditis immunology and pioneering new thinking for it.Methods:90 healthy female Wistar rats fed for one week(weight:120±20g) were randomly divided into 6 groups:normal group(G1) 15;model group(G2):15;tripterygiumforrestii and thyroid tablets treatment group(G3):15;formular sanjiexiaoying group(G4):15;formular wenshenjianpi group(G5):15;combined group of formular sanjiexiaoying and formular wenshenjianpi (G6):15.G1 with common feed;G2 - G6 treatment:100mg pig' s TG was completely dissolved in 50ml double distilled water, and 10ml was thoroughly mixed with same volume isovolumetic complete freund's adjuvant into water in oil type,and gave subcutaneous injection 0.5ml as primary immunization on back, cavum abdominis,legs and neck of rats.Two weeks later,we thoroughly mixed pig' s TG 10ml with incomplete freund' s adjuvant 10ml as boosting immunization once a week combined with sodium iodide to feed the rats with water until the fifth week(1L distilled water with 0.64g sodium iodide).From the sixth week,to feed G2 with common feed and water until the tenth week;to feed G3 with 2.75g/kg.d thyroid tablets and tripterygiumforrestii tablets by gastric perfusion until the tenth week;to feed G4 using 20g/kg.d formular sanjiexiaoying by gastric perfusion until the tenth week;to feed G6 20g/kg.d formular sanjiexiaoying and formular wenshenjianpi by gastric perfusion until the tenth week.In eleventh week,the rats were killed and blood and the thyroid tissues were taken out with HE colouring and observed the structure of rat's thyroid tissues with microscope.Hepatic,kidney function and blood-routine with reagent box's show. TGAb,TpoAb,FT3,FT4,TSHwere tested by reagent box' s show,TNF-α, IL-6were tested by ELISA,thyroid cells CD40,Fas,FasL were tested by immunohistochemistry.Results:1.Blood serum TGAb and TpoAb of model group increased and had significant difference(P < 0.01) compared with the normal group.Blood serum TGAb and TpoAb of tripterygiumforrestii group reduced and had significant statistical difference(P < 0.05 ) compared with model group;Blood serum TGAb and TpoAb of formular sanjiexiaoying group and combined treatment group reduced and had significant statistical difference(P < 0.01) compared with model group;Blood serum TGAb and TpoAb of formular sanjiexiaoying group and combined treatment group reduced and had not significant statistical difference(P> 0.05) compared with tripterygiumforrestii group;Blood serum TGAb and TpoAb between the formular sanjiexiaoying group and combined treamtent group,their differences had no statistical significance(P> 0.05).Blood serum TGAb and TpoAb between the formular wenshenjianpi group and model group,their differences had no statistical significance(P> 0.05).In normal group complete endolymph was soaked backgroud's colloidwas abundant,their distributing is concentrated and flaking,among follicle blood vessel was plenty.follicular epithelium was not acidophilic changed.Folial structure of model group was kept;interstitial endolymph was soaked; follicular epithelium acidophilic changed;epithelial cells enlarged as columnar;cell volume was big;boundary was clear; caryon was big.We found the lymph tissue distributing in and around folial and formed folliculi lymphaticus having germinal center.normal group,model group,tripterygiumforrestii and thyroid tablets treatment group,formular sanjiexiaoying group, formular wenshenjianpi group,combined group of formular sanjiexiaoying and formular wenshenjianp,the soaked inters -titial endolymph was letter than model group.The degree and bound of lymph cell soaked was reduce significant.formular sanjiexiaoying group and combined group the soaked interstitial endolymph was letter than tripterygiumforrestii and thyroid tablets treatment group,formular wenshenjianpi group. And follicular epithelium acidophilic changed was more lighten than tripterygiumforrestii and thyroid tablets treatment group,formular wenshenjianpi group.We foundn' t the lymph tissue distributing in and around folial and formed folliculi lymphaticus having germinal center.2.Lymphocyte soakage degree of model group obviously increased and had significant statistical difference(P< 0.05 ) comparing with normal group.Lymphocyte soakage degree of tripterygiumforrestii group,formular wenshenjianpi group decreased and had significant statistical difference(P< 0.05 ) compared with model group.Lymphocyte soakage degree of formular sanjiexiaoying group and combined treatment group decreased and had significant statistical difference(P< 0.01) compared with model group.Lymphocyte soakage degree of formular sanjiexiaoying group and combined treatment group reduced and had significant statistical difference(P<0.05) compared with tripterygiumforrestii group.The differences between the lymphocyte soakage degree of formular wenshenjianpi group and tripterygiumforrestii group had no statistical significance(P > 0.05 ).3.Blood serum TSH content of normal group,model group,tripterygiumforrestii group,formular sanjiexiaoying group, formular wenshenjianpi group,combined group were no significant statistical diference(P> 0.05 ).Blood serum FT3, FT4 content of normal group,model group,tripterygiumforrestii treatment group,formular sanjiexiaoying were no significant statistical diference(P > 0.05 ).Blood serum FT3,FT4 content of formular wenshenjianpi group and combined group obviously decreased and had significant statistical statistical difference(P < 0.05) compared with model group.4.Before the two weeks of madding mode,the model group compaired with normal,the growth rate of rats' weight, Appetite and excreation had no statistical significance.From the third week,the growth rate of rats' weight,appetite and excreation of the model group's rat had decreased compaired normal group and had significant difference(P < 0.05 ),their activity amount were less.After intervened by medicine,compared with model group, formular wenshenjianpi group,formular sanjiexiaoying group and combined treatment group,the growth rate of rats' weight of formular wenshenjianpi group was obviously increased,rats' appetite and excreation also increased and their activity amount were frequent comparing with other groups.We found the rats appetite of tripterygiumforrestii group decreased and their activity amount were less might relate with the side effect of their gastrointestinal system.The rats' tempreture of model group obviously reduced and had significant difference(P < 0.01) compared with normal group.There were no statistical differences(P > 0.05 ) of the rats' tempreture between tripterygiumforrestii group,formular sanjiexiaoying group and model group.The animal heat of formular wenshenjianpi group and combined treatment group increased comparing with other groups,their differences had statistical significance(P < 0.05 ).Also we found the rats' activity and mental state of formular wenshenjianpi group, formular sanjiexiaoying group and combined treatment group better than model group.5.The hepatic function between model group and normal group had no statistical differences(P> 0.05).AST,BUN,Cr,WBC, RBC of tripterygiumforrestii group had significant difference (P< 0.05) compared with normal group.AST,BUN,Cr,WBC,RBC of formular wenshenjianpi group,formular sanjiexiaoying group and combined cure group had significant difference(P < 0.05) compared with tripterygiumforrestii group.Among f formular wenshenjianpi group,formular sanjiexiaoying group and combined cure group,AST,BUN,Cr,WBC,RBC had no statistical differences (P>0.05).6.Fas/FasL immune positive substance of normal group presented light brown-red color,the expression ranges were very small.Fas/FasL immune positive of model group presented brown-red color,high concentration and the expression ranges were extended,compared with normal group and OD value had significant differences(P<0.01)The color of tripterygiumforrestii group,formular sanjiexiaoying group,formular wenshenjianpi group and combined treatment group obviously faded and the expression ranges of them became smaller.The color of formular sanjiexiaoying group and combined treatment group faded the most observably compared with tripterygiumforrestii group and OD value had significant differences(P <0.05).OD value between formular wenshenjianpi group and tripterygiumforrestii group had no statistical differences(P > 0.05).OD value between formular sanjiexiaoying group and combined treatment group had no significant statistical difference(P>0.05).7.CD40 immune positive substance of normal group presented light brown-red color,the expression spectum was very small. CD40 immune positive substance of model group presented brown-red color,high concentration and the expression spectum was extended,compared with tripterygiumforrestii group and OD value had significant differences(P < 0.01).The color of tripterygiumforrestii group,formular sanjiexiaoying group,for-mularwenshenjianpi group and combined treatment group obviously faded and the expression spectum of them became smaller.The color of formular sanjiexiaoying group and combined treatment group faded the most observably compared with tripterygium-forrestii group and OD value had significant differences(P <0.05).OD value between formular wenshenjianpi group and tripterygiumforrestii group had no statistical differences(P > 0.05).OD value between formular sanjiexiaoying group and combined treatment group had no significant statistical difference(P> 0.05).8.We found blood serum TNF-αcontent of normal group was very small,blood serum TNF-αcontent of model group were more increased than normal group and had significant statistical differences(P < 0.01).After treatment,blood serum TNF-α,IL-6 content of formular sanjiexiaoying group,combined treatment group,tripterygiumforrestii group,formular wenshenjianpi,were more decreased than model group and had significant statistical differences(P< 0.01);among which, blood serum TNF-αcontent of formular sanjiexiaoying group and combined group were more decreased than tripterygiumforrestii group and formular wenshenjianpi and had significant statistical differences(P < 0.01);blood serum TNF-αcontent of formular sanjiexiaoying group,combined treatment group had no significant statistical difference(P> 0.05).9.We found blood serum IL-6 content of normal group was very small.blood serum IL-6 content of model group were more increased than normal group and had significant statistical differences(P < 0.01).After treatment,blood serum IL-6 content of formular sanjiexiaoying group,combined treatment group,tripterygiumforrestii group,formular wenshenjianpi, were more decreased than model group and had significant statistical differences(P<0.01);among which,blood serum IL-6 content of formular sanjiexiaoying group and combined group were more decreased than tripterygiumforrestii group and formular wenshenjianpi and had significant statistical differences(P < 0.01);blood serum IL-6 content of formular sanjiexiaoying group,combined treatment group had no significant statistical difference(P> 0.05).Conclusions:1.formular sanjiexiaoying and formular wenshenjianpi can reduce the formation of autoantibody TGAb and TPOAb of autoimmune thyroiditis,and adjust immune response.2.Dissipating and eliminating goiter recipe and formular wenshenjianpi can reduce lymphocyte soakage and lighten its' inflammatory response.At the same time,formular sanjiexiaoying has an obviously efficacy for shortening swelled thyroid which is better than formular wenshenjianpi.Formular wenshenjianpi can increase the appetite and the excretion.The basal body temperature and basal metabolism of rats were improved.Comparing with tripterygiumforrestii,formular wenshenjianpi can avoidrats' gastrointestinal reaction and improve the clinical symptoms of HT effectively.It has an obviously symptom of function decrease for improving rats autoimmune thyroiditis.The efficacy is better than formular sanjiexiaoying and tripterygiumforrestii.4.formular sanjiexiaoying and formular wenshenjianpi can protect rats' hepatorenal function and hematological system which is better than tripterygiumforrestii.5.formular sanjiexiaoying and formular wenshenjianpi both can restrain TNF-α,IL-6 over secreting,it indicates that dissipating& eliminating goiter recipe is better than triptery -giumforrestii.6 The over expression of T cell commom stimulated molecule CD40 as the second activated signal was relevant ofmechanism of autoimmune thyroiditis.Autoimmune thyroiditis can be arisen through Th2 route.Formular sanjiexiaoying and formular wenshenjianpi can effectively restrain the espression of CD40, decrease T cell over activation and reverse the thyroiditis.7.The apoptosis mechanism of thyroid epithelial cells is one of reasons that cause the pathogenesis of autoimmune thyroiditis.formular sanjiexiaoying and formular wenshenjianpi can effectively restrain the apoptosis thyroid epithelial cells,decrease over activation and reverse the thyroiditis.
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