| BackgroundGraves’ Disease (GD) with a higher prevalance (about1～2%) is one of the common autoimmune endocrine metabolic disease.In recent years, with the impact on people’s lives such as diet, feelings and environment of modern factors, there is a growing trend in Graves’ disease.GD is characterized by goiter and thyroid hormone overproduction, and make the patients’ physical and mental health severely affected.In TCM, this Disease has a close contact with feelings, because of Qi produce phlegm, and blood stasis caused by phlegm, the blood stasis and phlegm knot in part of the thyroid gland of the neck each other. The therapy of morden medical science with a high cost, a big risk and high recurrence ratemainly rely on operation or anti-thyroid medication. Thyroid gland goiter persist is a severe problem of treatment. Resolving phlegm to dissipate stagnation and eliminate ying is a core and fundamental of the treatment in traditional chinese medicine.It has a certain characteristics and advantages in the therapy of goiter and reduction of the recurrence rate.At present, there is an opinion that this disease is an autoimmune disease, and the key of the morbidity is the product of Thyrotropin receptor antibody (TRAb) in the case of immune stimulation of body.lt stimulates directly thyroid receptor in the thyroid cells to produce thyroid hormone, in the same time it promotes the activation of iodine and then aggravates the T3, T4levels rise. T3, T4is the sign of thyroid function, so in the centre of TRAb, the correlational studies based on the treatment mode of resolving phlegm to dissipate stagnation in TCM is the focus of our research. The subject comes from the National Natural Science Foundation Project "phlegm method of Graves’ disease in a mouse model of thyroid hormone synthesis big impact downstream events.Early in phlegm medicine centered research found that it can significantly affect the thyroid cell proliferation and apoptosis, phlegm FRTL medicine can inhibit cell proliferation and induce apoptosis in thyroid cells expressing the Bax gene, which effects of drugs and other treatment of goiter similar mechanism; through the proliferation/apoptosis, the core factor study found upstream of goiter Consumers can upregulate the expression of apoptotic stimuli factor USF2mRNA, prompting a positive regulator of cell proliferation and cyclin B1, Ki67expression down to make a negative regulator of P21, P27upregulated; ultimately mediated pro-apoptotic genes BAX, FAS expression, induction of apoptosis, thereby improving the balance between dynamics of thyroid cell proliferation/apoptosis; the study also found that cells expressing phlegm medicine can TPO, thyroglobulin TG significantly reduced.In recent years, with the emergence and development of GD animal models, the treatment of resolving phlegm to dissipate stagnation influence on the pathological mechanism caused by TRAb subjects to explore. Iodine is raw material for the synthesis of thyroid hormones, TPO is a key synthetase thyroid hormone and TG is related to the storage protein, we have an opinion that rise of hormone levels with the activation of iodine are closely related, also Iodine storage is the key to goiter. Our group has been successfully build this model, and studied on thyroid hormone synthesis related factors, we found our traditional Chinese medicine of resolving phlegm to dissipate stagnation Inhibit the expression of TPO and TG. Therefore, in order to further the exploration on the mechanism of this aspect.This study is targeted at non-marine herbs of resolving phlegm to dissipate stagnation, and the mice model of GD immunized by Ad-TSHR289/6xHis as carriers. Through determination of the TRAb T3, T4and thyroid tissue, detection of activity of thyroid peroxidase and the content of measurement of thyroglobulin to observe the thyroid tissue function form, the key synthase of thyroid hormones and Iodine storage.ObjectiveStudy the thyroid function and morphology of GD mouse model treatwith the phlegm medicine, and explore the activation and storage of iodine after this TRAb produced.MethodsThe GD animal models induced by Ad-TSHR289/6xHis adenovirus immunity6-8week old female BABL/c mice.Treat the GD animal models with large dose of traditional Chinese medicine,low-dose of traditional Chinese medicine the western drug of methimazole and detect the different group of serum thyroid hormone T3, T4and its autoantibodies TRAb levels by radioimmunoassay detection methods; simultaneous watch the thyroid tissue weight of each group and HE staining of thyroid tissue; using the guaiacol method for the determination of each group TPO activity in thyroid tissue,and explain the phlegm medicine whether for effecting the activation of iodine; using radioimmunoassay detection of serum TG levels for phlegm medicine effect the storage of iodine.Results1.Thyroid function and morphology of Graves’disease animal models treated with phlegm medicine.1.1Successfully constructed Ad-TSHR289/6xHis adenovirus:its titer is3.16x1010PFU/ml.1.2Successfully build GD animal models:the incidence hyperthyroidism was70%,and the positive rate of TRAb was70%.1.3Thyroid functionThe most obvious difference was significant (P<0.05) high-dose group and western medicine group T3levels down GD mouse model with the control group Chinese high-dose group, low-dose group and western medicine group compared to the untreated group T3levels decreased44.8%, respectively,23.6%,35.3%(P <0.05), while the high-dose group and western medicine group T3levels were close to the normal group, quite effective, less effective and low-dose groupT4levels than the normal group after molding T4content of30%, after administration of normal T4levels reduced by14.4%, the model group decreased by13%, while the model treatment group T4levels were significantly reduced which traditional Chinese medicine to reduce43.9%of the high dose group Chinese medicine to reduce the low-dose group and western medicine group, only44.8%decreased21.8%. After the model in vivo administration of T4levels were lower in mice. in which the Medicines administered after T4levels consistent with the normal group, high-dose and low-dose medicine group did not significantly different, so the effect of the Medicines administered better;After molding TRAb levels higher than normal group was88.7%, after administration of each group to detect serum TRAb, wherein:the control group decreased by47.9%,27.3%in the untreated group, the model of traditional Chinese medicine fell83.9%in the high dose group, the model group decreased61.6small dose of medicine%,55.5%of western medicine group, the largest decline in Chinese high-dose group, and close to the normal group, and therefore the effect of high-dose group of Chinese medicine is more obvious1.4thyroid tissueThe Net-Weight of thyroid tissue:the weight of GD mouse model of thyroid tissue,treatwith high-dose of traditional Chinese medicine,were reduced by84.4%,which is most obvious,significant difference (P<0.05) contrast the control group. The western group were only decreased by21.8%(P<0.05), and after treatment of thyroid medication group, the Net-Weight of thyroid tissue was still higher than the normal group, after therapy of low-dose,the thyroid weight appeared to increase,and the standard deviation is larger, the changes were unstable, so high-dose of traditional chinese medicine in the treatment of goiter areas has obvious effect.The pathological morphology of thyroid tissue:macroscopic:the thyroid of model group and western medicine group were congestive and enlargement, which was the shuttle whip-like, and the thyroid treatment of phlegm medicine group were restored to light red; light microscope:the thyroid follicular enlargement and and edema of high-dose traditional chinese medicine intervention were become lighter,the proliferation of thyroid epithelial cell was significantly reduced, to improve the effect of goiter.2The downstream events of thyroid hormones synthesis after the Graves’ disease mouse model treatwith Phlegm fights method2.1TPO activityThe TPO activity of model group increased75.8%than normal group(P<0.05), traditional chinese medicine group was decreased8.85%(P<0.05),and western group decreased by40.1%, which is a big margin and may affect normal thyroid function, so the effect traditional chinese medicine group is quite excellent.2.2serum TG levels After treatmentNormal group differences before and after treatment was not significant (p>0.05), model group and western medicine group were significantly increased〠Chinese Medicine, significantly decreased low-dose group, model group and western medicine group increased, respectively:27.6%,22.6%(P<0.05), a large Chinese medicine, low-dose group decreased by34.7%,23.5%(P<0.05).Conclusions1Successfully established the mouse model of Graves’disease;2phlegm medicine significantly reduce thyroid hormone (T3, T4) and TRAb levels of Graves’disease mouse model, and reducing the net weight of thyroid tissue, significantly improved the organizational structure of thyroid enlargement;3phlegm medicine can significantly improve thyroid peroxidase (TPO) activity, and accelerating the activation of iodine; phlegm medicine can reduce the effect of TG, to improving its enlargement of thyroid tissue. |
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