Professor Wei Junping's Experience In Staged Treatment Of Hashimoto's Disease And Research On The Mechanism Of Action Of Xiapu Xiaoying Granule

Background:Hashimoto’s thyroiditis(HT)is a common autoimmune thyroid disease,characterized by significantly elevated levels of TPOAb in patients.In recent years,the incidence of this disease has gradually increased.Current research suggests that this disease is mainly the result of the interaction between genetic factors,autoimmune factors,and environmental factors,but the specific pathogenesis has not yet been fully elucidated.Therefore,there is a lack of targeted treatment methods in clinical practice,and most people with normal thyroid function and only elevated antibody levels are still in the passive observation stage until the condition develops into hypothyroidism,which to some extent affects the quality of life of patients.Therefore,finding effective interventions in the early stages of HT has important clinical significance.As an important part of the medical system,traditional Chinese medicine(TCM)has a unique theoretical system and significant therapeutic advantages.Objective:1 To explore the main treatment methods and medication patterns of Professor Wei Junping in the early,middle,and late stages of treating HT.2 To analyze the cumulative transition probability of patients’ states at different times(Markov cycle)after receiving the guidance of "using Banxia Houpu Decoction combined with Xiaoyao Powder as the basic formula for early HT,using Liu Junzi Decoction combined with Jujube and Platycodon Decoction as the basic formula for intermediate HT,and using Liuwei Dihuang Pill as the basic formula for late HT",predict the disease dynamics and long-term efficacy,and evaluate the efficacy of the above treatment methods for HT.3 Observe the effect of Xia Po Xiao Ying Granule,derived from the mentor’s experience,on the thyroid function and morphology of EAT rats constructed by immune-mediated combined with high iodine method,providing pharmacodynamic evidence for further development and application of traditional Chinese medicine prescriptions.4 To explore the mechanism of Xia Po Xiao Ying Granule in intervening in the necroptosis of thyroid cells by regulating the TLR4/RIPK3/MLKL signaling pathway.Methods:1 This study used the Chinese medicine prescriptions for treating HT during different stages,which were provided by Professor Wei Junping between January 2011 and December 2020,as the research data.The Ancient and Modern Medical Case Cloud Platform was employed,and data mining techniques were utilized to explore the treatment methods and medication rules of the tutor in treating HT in the early,middle,and late stages.2 Taking HT patients as the research subjects and changes in disease progression as the endpoint event,based on the basic principles and methods of Markov model,HT outcomes were divided into four different Markov states,and the transition probability between each Markov state was calculated.The Tree Age Pro software was used to construct a multi-state Markov decision tree model,with 12 weeks as one Markov cycle.The cumulative transition probability of patients in different Markov states after receiving treatment for 10 and 20 Markov cycles was analyzed to predict disease dynamics and long-term efficacy.3 The EAT rat model was constructed by immune-mediated combined with high iodine method.The serum levels of TSH,FT3,FT4,TT3,TT4,TPOAb,and TgAb were detected by ELISA.The pathological morphology of the thyroid tissue was observed under light microscopy using HE staining,and the thyroid inflammation score was evaluated.The effect of Xia Po Xiao Ying Granule on the thyroid function and morphology of EAT rats was evaluated.4 qPCR was used to detect the relative expression levels of TLR4,MLKL,and RIPK3 genes.Western Blot was employed to detect the protein expression levels of TLR4,MLKL,and RIPK3.Immunohistochemistry was used to detect the expression of MLKL in thyroid tissue.The mechanism of Xia Po Xiao Ying Granule in intervening in necroptosis of thyroid cells was explored.Results:1 The analysis of complex networks shows that the core prescriptions used by the mentor to treat HT in the early stage are:Pinellia ternata,Magnolia officinalis,Poria cocos,Bupleurum chinense,Atractylodes macrocephala,Citrus reticulata.Banxia Houpu Decoction combined with Xiaoyao Powder as the basic formula for treating hypothyroidism;the core prescriptions used in the intermediate stage are:Poria cocos,Pinellia ternata,Citrus reticulata,Licorice root,Platycodon grandiflorus,Chinese wolfberries.Liu Junzi Decoction combined with Jujube and Platycodon Decoction as the basic formula;and the core prescriptions used in the late stage are:Chinese wolfberries,Poria cocos,Cortex Moutan,Cyathula root,Rehmannia root.Liuwei Dihuang Pill as the basic formula.The results suggest that the early treatment of HT should be focused on soothing the liver and resolving depression,regulating qi and resolving phlegm.In the intermediate stage,the focus should be on strengthening the spleen and drying dampness,resolving phlegm and dissipating masses.In the late stage,the focus should be on nourishing the liver and kidneys,strengthening the spleen and promoting yang.2 HT patients who received treatment guided by "Banxia Houpu Decoction combined with Xiaoyao Powder for the early stage,Liu Junzi Decoction combined with Jujube and Platycodon Decoction for the intermediate stage,and Liuwei Dihuang Pill for the late stage" showed the following outcomes after 10 and 20 Markov cycles:patients who were initially diagnosed with normal thyroid function but had elevated thyroid antibodies remained in this state for the most part;patients who were diagnosed with hypothyroidism at the beginning but received consistent treatment had a certain chance of reversing this condition;and patients who were initially diagnosed with hyperthyroidism or subclinical hypothyroidism but received treatment for a period of time had a high probability of converting to normal thyroid function.3 Both male and female rat models showed elevated TgAb and TPOAb levels compared to the control group(P<0.001).After intervention with Xia Po Xiao Ying Granule,both groups showed a downward trend.HE staining results showed that the thyroid follicles in the control group and the control group with medicine were relatively uniform in size,with intact structure,and the follicular lumen was filled with colloid and a varying number of "absorption vacuoles"of different sizes,without significant inflammatory cell infiltration.In the model group,thyroid follicles of different sizes were observed,with different degrees of atrophy,mild expansion of follicular spaces,and varying degrees of lymphocytic infiltration.After treatment with various drugs,the degree of thyroid injury improved to varying degrees,with the most significant improvement observed in the high dose group and the positive control group.4 The relative expression levels of TLR4,MLKL,and RIPK3 genes in the model group were upregulated compared to the control group.Compared to the model group,the relative expression level of TLR4 gene in the low and high dose groups and the positive control group showed a downward trend,while the relative expression levels of RIPK3 and MLKL genes in all Chinese herb dose groups showed a downward trend.Conclusion:1 In the early stage of HT,the liver loses its regulatory function of discharging and there is an interaction between phlegm and qi.Therefore,the main treatment method should be to regulate the disharmony of the liver and resolve stagnation,as well as to regulate qi and transform phlegm.The Ban Xia Hou Po Decoction combined with the Xiaoyao Powder should be used as the basic formula with modifications.In the intermediate stage,the liver qi invades the spleen and the spleen is weakened due to dampness.The main treatment method should be to strengthen the spleen,dry dampness,resolve phlegm,and dissipate masses.The Liu Jun Zi Decoction combined with the Jie Geng Ban Xia Decoction should be used as the basic formula with modifications.In the late stage,there is deficiency in the liver,spleen,and kidneys.The main treatment method should be to nourish the liver and kidneys,strengthen the spleen,and assist yang.Medications should be based on the Liu Wei Di Huang Wan with modifications.2 Patients who receive treatment guided by the plan of "using Ban Xia Hou Pu Decoction combined with Xiaoyao Powder in the early stage,Liu Jun Zi Decoction combined with Jie Geng Ban Xia Decoction in the intermediate stage,and Liu Wei Di Huang Wan in the late stage"can achieve good long-term therapeutic effects.Early intervention can prevent disease progression,promote thyroid function recovery,and prevent further development of the disease into hypothyroidism.3 Xia Po Xiao Ying Granule can effectively reduce serum TgAb and TPOAb levels in both male and female SD rats with EAT induced by immune-mediated combined with high iodine method.It can also lower thyroid pathological scores,and improve thyroid damage in EAT rats.4 The injury of HT thyroid cells may be related to the activation of the TLR4/RIPK3/MLKL signaling pathway,and the protective effect of Xia Po Xiao Ying Granule on thyroid tissue in EAT rats may be related to the reduction of the expression of the key protein MLKL,which is involved in necroptosis.