| Objective:Hashimoto’s thyroiditis(HT)is the most common autoimmune thyroid disease(AITD).The pathogenesis of HT is complex and the recognized treatment method is deficient,currently.Traditional Chinese medicine(TCM)has certain advantages in HT.Therefore,this study aims to observe the clinical efficacy and safety of Qinggansanjiexiaoying Decoction in the treatment of HT through clinical trials.Proteomics analysis was carried out to explore the potential therapeutic mechanism of Qinggansanjiexiaoying Decoction.Verifying the potential proteins from the proteomics was performed to explore the potential therapeutic targets of Qinggansanjiexiaoying Decoction,and provide scientific connotation for the treatment of HT with TCM.Methods:In the first study,a non-randomized controlled trial method was applied to recruit Hashimoto’s thyroiditis patients from the department of thyroid disease,Dongzhimen Hospital and Sunsimiao Hospital.Beijing University of Chinese Medicine.The patients were divided into two groups according to the ratio of 1:1.The control group was given a limited iodine diet,and the experimental group was given Qinggansanjiexiaoying Decoction orally combined with a limited iodine diet.The course of treatment in both groups was 4 weeks.The serum TPOAb titer,TGAb titer,TSH level,thyroid function,routine blood test,liver and kidney function,using ultrasound to measure and calculate thyroid volume,using the Thy PRO-3 9 scale to evaluate the patient’s quality of life and TCM syndrome integration scale to evaluate the improvement of TCM syndrome before and after treatment,respectively.In the second study,5 patients were randomly selected from each group of the experimental group and the control group in the first study.The serum sample of selected patients were used as subjects.TMT labeled proteomics technology was performed to identify and quantitatively analyze the serum proteins of subjects based on UniProt database,screened the differential proteins related with the Qinggansanjiexiaoying Decoction before and after treatment,and carried out protein function and pathway annotation,enrichment analysis and protein interaction analysis through bioinformatics analysis.In the third study,differential proteins related to the mechanism of Qinggansanjiexiaoying Decoction in treating HT were selected from the differential proteins in the second study.Serum samples of subjects before and after treatment were randomly selected from the experimental group and the control group in the first study.The expression levels of candidate differential proteins in the experimental group and the control group before and after treatment were detected using the ELISA method,indicating the direction for future research.Result:In the first study,a total of 109 patients were included in the clinic study.There were 54 patients in the experimental group,with a female/male ratio of 46/8 and an average age of 38.50(31.75,51.00)years old.There were 55 patients in the control group,with a female/male ratio of 52/3 and an average age of 37.00(29.25,50.75)years old.There was no statistically significant difference in gender,age,BMI,course of disease between the two groups(P>0.05).The experimental group showed a significant decrease in the level of TPOAb after treatment(average of decrease rate was 9.24%,P=0.003<0.01),while the control group showed a slight increase(average of increase rate was 3.99%,P=0.235>0.05).The serum level of TGAb decreased about 3.77%(median)in the experimental group,while increased about 0.72%(median)in the control group,none of them showed a significant difference(P>0.05).The volume of the right lobe,left lobe,and isthmus of the thyroid gland significantly declined about 3.05%、4.16%、0.94%(P=0.009,0.001,0.034<0.05)after treatment in patients from the experimental group.In terms of quality of life,symptoms associated with thyroid goiter,fatigue,cognitive complaints,anxiety,depression,emotional fluctuations and the whole negative influence in the experimental group were significantly improved after treatment(P<0.05),but no significant improvement in the control group.The TCM syndrome showed a significant decline approximately 14.58%(P=0.022<0.05)in the experimental group,while no significant changes in the control group.The levels of TSH did not change significantly in the both groups(P>0.05).Thyroid function,routine blood test,liver and kidney function were remaining within normal range.In the second study,653 proteins were identified by TMT proteomics technology,and 58 potential differential proteins related to the efficacy of Qinggansanjiexiaoying Decoction were screened,including 14 up-regulated proteins and 44 down-regulated proteins.Through protein functional annotation,it was found that candidate differentially expressed proteins mainly participate in biological processes such as inflammatory response,immune response,cell and tissue structure formation,intercellular signaling,oxidative stress,cell apoptosis,and substance metabolism.Through KEGG pathway analysis,it was found that differential proteins mainly participate in the pathogenesis of HT pathways such as neutrophil extracellular trap network,IL-17 signaling pathway,and cell apoptosis pathway.Protein interaction analysis revealed that differential proteins such as Apolipoprotein E(APOE),serum amyloid A protein(SAA),serum amyloid P-component(APCS or SAP),and protein S100-A9(S100A9)may be core proteins that exert biological functions,and further experimental verification is needed.In the third study,a total of 7 differential proteins related to the pathogenesis of HT were selected from proteomics results,including S100A9,SAA,SAP,APOE,CNNM2,DMKN and LRG-1,which were related with immune inflammatory response.The ELISA test results showed that the serum levels of S100A9,SAA,SAP,APOE,CNNM2,DMKN,and LRG-1 in the experimental group were significantly reduced after treatment(P<0.05),the average decline rates were 9.61%,10.28%,13.76%,12.78%,4.55%,8.37%and 5.82%,respectively.However,there was no significant change in the levels of serum candidate proteins in the control group after treatment(P>0.05).The results from ELISA test were consistent with proteomics.Conclusion:The Qinggansanjiexiaoying Decoction can effectively reduce the serum levels of TPOAb of HT patients,reduce thyroid volume,improve symptoms such as thyroid goiter,fatigue,cognitive complaints,anxiety,depression and emotional fluctuations,and improve TCM syndrome conditions.It has significant efficacy and good safety,which is worth promoting and applying in clinical practice.The potential mechanism of Qinggansanjiexiaoying Decoction in the treatment of HT is mainly related to reducing immune inflammatory response,which involves multiple pathways such as IL-17 signaling pathway,NET pathway,and cell apoptosis.The Qinggansanjiexiaoying Decoction may regulate the immune inflammatory response of the body by reducing the levels of S100A9,SAA,SAP,APOE,CNNM2,DMKN and LRG1,and therefore play a therapeutic role in HT.It provides a new research direction for the diagnosis and treatment of HT and is worthy of further experimental research to verify. |
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