Mechanisms Of The Treatment Of Kangyan Xiaoying Recipe On Autoimmune Thyroiditis

Autoimmune Thyroiditis (AT), also known as chronic lymphocytic thyroiditis. It may be caused by genetic, endocrine, iodine, physics and chemistry factors, so that occurred in patients with immune function disorders.Result in a series of autoantibodies against thyroid tissue, leading to thyroid showed diffuse lymphocytic infiltration, fibrosis, interstitial atrophy and acinar cells in eosinophilic change. Hashimoto's statistics of 1619 cases of this disease, found in addition to clinical and pathological manifestations of goiter, debilitation was its primary symptoms. Pathology essence of primary disease marked asthenia in origin and asthenia in superficiality. Therefore, this temporary solution to Kangyan-based disease and to reduce inflammatory cell invasion, the main root of the problem with brooding gallae and suppression of the autoimmune response and antibody expression.Traditional Chinese medicine Tripterygium, ephedra had immunosuppressive action. They may different degrees of inhibit lymphocyte to response to antigens, thereby reducing autoimmune damage. Peach kernel with the cinnamon, had promoting blood flow and reinforce Yangqi effect in deficiency of vital energy and blood stasis in early disease. Clinical application of its own thyroiditis was found in patients with a certain therapeutic effect. Preliminary experiments were also found in a variety of Tripterygium ingredients and ephedra polysaccharide can reduce the expression of thyroid antibodies, can inhibit the hyperactivity of the immune response.On this basis, we have according to the principle of compatibility to compose Kangyan Xiaoying Recipe, observe effect in autoimmune thyroiditis antibodies and hormones expression, level of lymphocytes and its subsets and apoptosis-inducing factor in thyroid tissue. At the same time explore the mechanism of drug action, to provide a basis for the compatibility of drugs. Throughout the study in four partsThe first part Study multi-point injection of antigen plus iodine-induced experimental autoimmune thyroiditis (EAT) model to foundDivided 18～22g mice randomly into normal group and model group. Model group, each mouse was injected subcutaneously with pig thyroglobulin (PTg) complete Freund's adjuvant antigen were 0.2ml, for 2 weeks, once a week.3rd week of each mice were injected subcutaneously PTg incomplete Freund's adjuvant antigen 0.2ml,3 consecutive weeks, once a week. Iodine per day for drinking water. The general state of experimental mice was observed, changes in body weight and food intake was recorded. End of the experiment to take thyroid tissue in the rat, were observed under optical microscope histopathological changes. Specimens from serum samples by RIA detection of thyroid antibodies and hormone levels. Results:From the second week, the model group reduced food consumption, experiments from the fourth week, the model group animals, weight loss, began to emerge, inactivities, exophthalmos, hair erect. Experiment at the end, the model mice, body weight and food intake in serious decline. Pathological examination showed that model group thyroid to take on different degrees of acinar expansion, acinar colloid thin. Some acinar destruction, alveolar and interstitial lymphocyte infiltrated clearly, serious cases formed lymphoid follicles, it is thus clear that more eosinophil infiltration. Radioimmunoassay test results, the model group serum levels of hormones and antibodies increased. These results suggest that multi-point injection of antigen plus iodine can lead to serum TGAb and TMAb auto antibody high-titer expression: Thyroid follicular epithelial cells in eosinophilic change, necrosis and a large number of lymphocyte infiltration. Lymphoid follicles can be seen and was accompanied by the formation of germinal centers, also caused similar to hyperthyreosis symptoms such as weight loss, inactivities,exophthalmos, hair erection, etc.. Similar to the clinical AT very. Be used as an effective EAT model for evaluation of the role of drug-related.The second part Study Kangyan Xiaoying Recipe effects of hormone and antibody levels on the EAT mice.The first section used multi-point injection of antigen plus periodate to build autoimmune thyroid method, the same time took lOmg/kg body weight of drug-treated by intragastric administration, to observe effects of the Kangyan Xiaoying Recipe to thyroid tissue in mice. Results:The Kangyan Xiaoying Recipe repaired most of the thyroid tissue, reduced lymphocyte infiltration. Second section used radioimmunoassay to detect effects of Kangyan Xiaoying Recipe in the EAT serum thyroid hormone and antibody. The results showed that:model group, T3, T4, FT3, FT4, TGA, TMA levels were higher than the normal group, there was significant difference (p<0.01). Kangyan Xiaoying Recipe large dose group T3, T4, FT3, FT4, TGA, TMA levels are lower than model group, there are significant differences (p <0.01, p<0.05). Kangyan Xiaoying Recipe low dose group T3, T4, FT3, FT4, TGA, TMA lower than the model group, but no significant difference, so we think that Kangyan Xiaoying Recipe treatment of EAT effects and drug dose was into a certain correlation.The third part Study Kangyan Xiaoying Recipe effects of lymphocytes on the EAT mice.The first section used MTT assay effects of Kangyan Xiaoying Recipe in spleen cell proliferation of multi-point injection of antigen plus iodine-induced EAT mice model. Results show that:EAT model group of spleen lymphocyte proliferation significant higher than the normal control group, treatment group could significantly inhibit ConA induced spleen lymphocyte proliferation. The second section by flow cytometrey observed Kangyan Xiaoying Recipe effect on lymphocyte subsets CD4 +/CD8 + of EAT mice. Results showed that:model group, CD4 + lymphocytes/ CD8 + lymphocytes higher than the normal group, there were significant difference (p<0.01), shows the model group the number of lymphocyte subsets abnormal and balance disorders. CD8 + lymphocytes in the treatment group higer than in model group, there were significant difference (p<0.01); CD4 + lymphocytes/CD8 + lymphocytes lower than the model group, there were significant difference (p<0.01), shows Kangyan Xiaoying Recipe can improve the the relative content of CD8 + lymphocytes and lower CD4 + lymphocyte/CD8 + lymphocyte ratio, so that excessive immune system response has been effectively controlled.The fourth part Research on the Kangyan Xiaoying Recipe effects of thyroid apoptosis on EAT mice.The first section used by immunohistochemistry method to detect the influence of Kangyan Xiaoying Recipe on thyroid tissue vascular endothelial growth factor (VEGF) of multi-point injection of antigen plus iodine-induced EAT mice. The results showed that, EAT mice thyroid tissue VEGF was higher than the normal control group,but no significant difference.we believed that it related with the formation of EAT mice thyroid tissue lymphoid follicles, high expression of VEGF further promoted the formation of capillaries in the tissue, but the degree of its capillary formation differented to another autoimmune thyroid disease Graves. Believed it can distinguish two disease as a method.The second section used immunohistochemical staining method to detect the influence of Kangyan Xiaoying Recipe on multi-point injection of antigen plus periodate EAT model mice induced by apoptotic factor Fas and its ligand Fas-L in thyroid tissue. Results show that EAT mice apoptosis factor Fas and its ligand Fas-L in thyroid tissue showed high expression, Fas/Fas-L expression increased thyroid apoptosis, resulted in autoimmune phenomena. The treatment group significantly decreased Fas/Fas-L the high expression, thereby inhibited apoptosis to reach treatment of autoimmune thyroiditis.Conclusion:The multi-point injection of antigen plus periodate method was successful, be used as an effective EAT model, used for evaluating the role of drug-related. Kangyan Xiaoying Recipe can improve the state of EAT mice, correct thyroid hormone and antibody levels, inhibit lymphocyte proliferation, balanced the proportion of CD4 + lymphocytes and of CD8 + lymphocytes, reduce the VEGF and Fas/Fas-L in thyroid tissue high-expression.