The Beneficial Effects And Mechanism Of Four Chinese Herbal Formula In The Treatment Of Hypothyroidism

Objective: First,the effects and mechanism of four Chinese herbal formula to hypothyroidism-induced by chemical and experimental autoimmune thyroiditis(EAT)were explored.Second,the clinical therapeutic efficiency of C hinese herbal formula 1 were evaluated.Third,the possible target of Yin Ju San Jie Granules database was identified by Cellular Thermal Shift Assay(C ETSA).Thyroid Stimulating Hormone Receptor(TSHR)agonists among Yin Ju San Jie Granules database were screened by ligand-based pharmacophore model.Method: 1.Extraction of Chinese herbal formula Chinese herbal formula were extracted with 60% ethanol.2.The effects of Chinese herbal formula to hypothyroidism-induced by chemical The hypothyroidism model was induced by 0.1% propylthiouracil(PTU)in female Kunming mice.The levels of two thyoid hormones were determined to evaluate the effects of C hinese herbal formula;Morris water maze study and rotarod test were carried out to determine the effect of Chinese herbal formula on PTU induced memory deficiency and fatigue,respectively.Body weight and thyroid pathological features were also observed.3.The effects of Chinese herbal formula to EAT The EAT model was induced by PTg(Porcine Thyrogiobulin),and the levels of thyoid hormone and thyroid pathological changes were selected to study the impact of Chinese herbal formula to EAT thyroid function;Thyroid Autoantibody(TPOAb)and thyroglobulin antibodies(Tg Ab)were determined to study the impact of C hinese herbal formula to immune regulation;TNF-α,IL-10 and TGF-β1 serum levels were examined to evaluate the impact of Chinese herbal formula to the balance of Th1/Th2 cytokines.The levels of thyroid-stimulating hormone(TSH),Free Thyroxine(FT4)and FreeTriiodothyronine(FT3)as well as thyroid autoantibody TPOAb and Tg Ab were determined to evaluate the therapeutic effects of C hinese herbal formula in the treatment of patients with Hashimoto thyroiditis(HT).4.The study of the possible target and active components of Chinese herbal formula It was validated that whether the herbs of Yin Jusan Granules database were targeted protein TSHR by CETSA.After a ligand-based pharmacophore model was established,TSHR agonists were identified using this pharmacophore screening procedure.Results: 1.Extraction of Chinese herbal formula The yields of extraction of formula 1,formula 2 and formula 3 were 21%,24% and 22%,respectively.2.The effects of Chinese herbal formula to hypothyroidism Symptom: the mice have characterized by hair loss and burnout.The symptom could improve after the Chinese herbal formula treatment.Concerning the levels of thyroid hormone and thyroid pathological changes,compared with the control group,the level of FT4 was reduced while TSH was increased in the model group(p<0.05);compared with the model group,the level of FT4 was increased and TSH was reduced in the administration groups.The effect of formula 1 high and low dose groups were changed significantly(p<0.05).Thyroid pathological changes results: compared with the control group,the model group thyroid follicular epithelial cell hypertrophy,part of the follicle fusion into a huge follicle.compared with the model group,the follicular epithelial cell swelling degree was reduced in each administration group.Hypothyroidism with brain damage by Morris water maze study: compared with the control group,the average latency of the model group increased.Compared with the model group,the average latency of the treatment group was reduced.The percentage of mice in each group arriving at the platform was analyzed.Compared with the control group,the percentage of the model group reaching the platformdecreased only a few percent.Compared with the model group,the percentage of the administration group reaching the platform increased.In contrast,the positive control drug Yinju Sanjie granule and F2 low dose increased significantly(p <0.05).Hypothyroidism symptom fatigue: compared with the control group,the running time of mice in model group was significantly decreased(p<0.05).Compared with the model group,each drug administration group had the effect of prolonging the running time.Among drug treatment groups,formula 1 and formula 2 treatment groups at high dose significantly reversed this phenomenon(p <0.05).3.The effects of Chinese herbal formula to EAT The levels of thyroid hormone and the thyroid autoantibod y: compared with the control group,the level of FT4 was reduced while TSH,TPOAb and Tg Ab were obviously increased in the model group(p<0.05);compared with the model group,the level of FT4 was increased while TSH,TPOAb and Tg Ab were reduced in the drug administration groups.The effects of formula 1 and formula 3 groups at high dose were changed significantly(p<0.05).Thyroid pathological changes results: compared with the control group,the degree and range of lymphocyte infiltration in the model group was significantly increased(p<0.05).Compared with the model group,the degree and the range of lymphocyte infiltration decreased in administration group(p<0.05).Among them,the lymphocyte infiltration of formula 1 and formula 3 high dose groups decreased significantly(p <0.05).The levels of Th1/Th2 cytokines: compared with the control group,the level of IL-10 was reduced while TNF-α was increased in the model group(p<0.05);compared with the model group,the level of IL-10 was increased while TSH,TNF-α were reduced in the administratio n groups.The effect of formula 1 and formula 3 high dose groups were changed significantly(p<0.05).The expression of TGF-β1,TNF-α,and IL-10 changes results: compared with the control group,the expression of TGF-β1,TNF-α were increased,while IL-10 was reduced in the model group(p<0.05);compared with the model group,the expression of TGF-β1,TNF-α,were reduced,while IL-10 was increased in the administration groups(p<0.05).The effects of formula 1 and formula 3 high dose groups were changed significantly(p<0.05).4.A study of the possible target and active components of Chinese herbal formula 37 TSHR ligands were collected as a training set,we established a compound database including 733 compounds from the herbs of Yin Ju San Jie Granules,and then we got a model comprising four pharmacophore features and a pharmacophore volume constraint.The relevance ratio of agonists was 50% and GH Score of model was 0.875.According to the ligand-based pharmacophore search,26 compounds were found among the compound from Yin Ju San Jie Granules.Conclusion: 1.Chinese herbal formula could improve the symptoms of PTU-induced hypothyroidism.2.Chinese herbal formula could reduce hypothyroidism with brain damage and fatigue.3.Chinese herbal formula could reduce the autoantibodies Tg Ab and TPOAb formation.4.Chinese herbal formula could reduce the infiltration of lymphocytes in EAT.5.Chinese herbal formula could effectively regulate the immune function of EAT mice,reversing the imbalance of Th1 / Th2.6.The clinical therapy rate of formula 1 was 60%.7.Extract of Yin Ju San Jie Granules had interaction with TSHR protein.8.After ligand-based pharmacophore model was established,26 compounds were found among Yin Ju San Jie Granules database.These compounds would be further verified and identified to be TSHR agonists by experimental assay.