| Background:Autoimmune thyroiditis(AIT)is a very common autoimmune disease of the endocrine system,which is characterized by inflammatory infiltration of lymphocytes and elevation of autoimmune antibodies,mainly Thyroglobulin antibody(Thyroglobulin antibody,AIT).Tg Ab)and Thyroperoxidase antibody(TPOAb).Pyroptosis plays an important role in the pathogenesis and progression of AIT.NLR Family,Pyrin Domain Containing Protein 3(NLRP3)is one of the important upstream activators of pyroptosis,but there are few studies on its role in AIT.Professor Gao Tianshu,the tutor of AIT,believes that spleen deficiency is the root of AIT.In clinical practice,Buzhong Yiqi decoction,which is used to invigorate qi and strengthen the spleen,is often used for the treatment of AIT,and the curative effect is very good.From the perspective of syndrome differentiation of traditional Chinese medicine(TCM),combined with the traditional Chinese medicine inheritance assistance platform,this study summarizes the clinical cases of Professor Gao’s treatment of AIT with instilling qi and instilling spleen,explores the experience in the differentiation and treatment of AIT and the rules of medication,and further studies the mechanism of Buzhong Yiqi decoction on NLRP3inflammasoma-mediated pyroptosis in AIT model mice through animal experiments.To provide scientific basis for the traditional Chinese medicine treatment of AIT.Research methods:1 Collect the clinical cases of Professor Gao Tianshu’s treatment of AIT,build a medical record database,and use the frequency statistics,data analysis,cluster analysis,association rule analysis,complex system entropy aggregation,entropy chromatography cluster analysis and other data mining methods of traditional Chinese Medicine Inheritance auxiliary platform(V2.5)to explore the drug characteristics and prescription rules of Professor Gao’s treatment of AIT.The clinical experience of tutors was summarized.2 Animal experiment: 72 SPF female NOD.H-2H4 mice were randomly divided into 6 groups:control group(ZC),model group(M),low-dose Buzhongyiqi decoction group(D),medium-dose Buzhongyiqi decoction group(Z),high-dose Buzhong Yiqi decoction group(G) and western medicine selenium preparation group(X),with 12 mice in each group.The normal group was fed with distilled water,and the other groups were fed with 0.05% sodium iodide(Na I).After the model was successfully established for 8 weeks,the mice were given intragastric administration once a day from the 9th week after the model was established,with9.1 times the daily dosage of human body weight(g/kg)as the standard.The degree of inflammatory infiltration in thyroid tissue was observed by HE staining.Serum Tg Ab levels were measured by ELISA.The expressions of NLRP3,caspase-1,IL-1β and IL-18 in thyroid tissues were detected by immunohistochemistry,Real-Time PCR and Western Blot.Results:1 A total of 98 prescriptions were included in the study,including 146 Chinese herbs,and the total frequency of use was 1320 times.Among them,a total of 20 herbs were used more than20 times,mainly warm and sweet herbs,and most of the meridians were spleen meridians.A total of 20 core drug combinations were used more than 25 times.The data were processed by complex system entropy clustering method,and 22 core drug combinations that could be used for new prescription clustering were finally obtained.2 Effect of Buzhongyiqi decoction on AIT model mice.2.1 Evaluation of model indicators: HE staining under light microscope showed that the inflammatory infiltration area of the thyroid of mice in each model group was greater than 2%of the total thyroid area.2.2 Evaluation of inflammatory cell infiltration: The degree of inflammatory cell infiltration in thyroid tissue sections stained with HE was observed.2.3 Detection of thyroid antibody TgAb: the level of TgAb in the model group was significantly higher than that in the normal group,and the level of antibody in each administration group was improved to varying degrees compared with the model group,but still higher than that in the normal control group.3.Effect of BuzhongYiqi decoction on NLRP3 inflammasome and pyroptosis in AIT model mice.3.1 Immunohistochemical results showed that NLRP3,caspase-1,IL-1β and IL-18 were expressed in the thyroid of each group.Compared with the normal group,the optical density values of the above indicators in the model group were significantly higher,and the optical density values of the above indicators in each drug group were lower than those in the model group.3.2 The results of Real-Time PCR showed that NLRP3,caspase-1,IL-1β and IL-18 m RNA were expressed in the thyroid of each group.Compared with the normal group,the m RNA expression of the above indicators in the model group was significantly increased.Compared with the model group,the m RNA expression of the above indicators in each treatment group decreased to varying degrees.3.3 The results of Western Blot showed that NLRP3,caspase-1,IL-1β and IL-18 proteins were expressed in the thyroid of each group.Compared with the normal group,the protein expressions of the above indicators in the model group were significantly increased.Compared with the model group,the protein expressions of the above indicators in each treatment group were decreased to varying degrees.Conclusions:1.The tutor mostly treats AIT with invigorating qi and invigorating spleen,and the curative effect is very good.Most of the prescriptions are composed of sweet,warm and spleen meridian drugs,and the treatment is based on the spleen,invigorating spleen and invigorating qi,so as to achieve the effect of AIT intervention.2.Buzhong-yiqi decoction can significantly improve the inflammatory state of thyroid in AIT model mice,and its mechanism may be achieved by NLRP3 inflammasoma-related pyroptosis. |
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