The Study Of Screening Serum Biomarkers Of Autoimmune Thyroid Disease By Protein Chip

BackgroundAutoimmune thyroid disease is the most common group of organ-specific autoimmune diseases,mainly including Graves' disease,Hashimoto's thyroiditis,which are mainly thyroid dysfunction caused by thyroid autoantibodies.Global epidemiology shows that its incidence is high,and more common in women,there is no obvious clinical realization in the initial stage,Most of them are discovered only after serious complications.Protein chip is a new powerful tool for proteomics research,which is developed on the basis of gene chip.It has special advantages,such as high throughput,integration and miniaturization.protein chip has two characteristics,On the one hand,as a semi-quantitative tool,it can simultaneously detect multiple cytokines in a sample and analyze the expression between samples;On the other hand,the relationship between genes and cytokines can be studied as a complement to gene chips.protein chip has been gradually recognized in the application of common clinical diseases,such as rheumatoid arthritis,tumors,cardiovascular diseases and other related reports,but there are few reports on the application of autoimmune thyroid diseases.PurposeProtein chip was used to detect the expression of cytokines in autoimmune thyroid disease patients,and the differentially expressed cytokines were selected by software and statistical analysis.For the selected cytokines,expand the sample and use enzyme-linked immunosorbent assay for quantitative detection,perform statistical analysis to obtain statistically significant cytokines,and explore the possible role of cytokines in autoimmune thyroid disease patients.MethodsSerum samples from 5 patients with autoimmune thyroid disease patients(experimental group)and 3 healthy controls(control group)were prepared for highthroughput cytokine screening by protein chip technology.After screening the differentially expressed cytokines by protein chip,Enlarged sample of 64 cases,41 cases in the experimental group,23 cases in the control group,using ELISA to verify the experiment,the expression of cytokines quantitative detection.According to the normal distribution,t-test or rank sum test was used to get the statistically significant cytokines in the two groups.Results1.Detection of cytokines by protein chip technology: The protein chip included 80 cytokines,and four cytokines were screened,which were pulmonary activation regulated chemokine,insulin-like growth factor binding protein 3,thrombopoietin,vascular endothelial growth factor,in which insulin-like growth factor binding protein 3was down-regulated and the rest were up-regulated.2.Enzyme-linked immunosorbent assay(ELISA): Enlarge the sample size,the relevant cytokines screened by protein chip were detected by ELISA,and the relevant results of cytokines expression were analyzed statistically.The results showed that compared with the control group,the expression of insulin-like growth factor binding protein 3 in the experimental group was higher(p > 0.05);The expression of pulmonary activation regulated chemokine in experimental group was lower than that in control group(p > 0.05);The expression of thrombopoietin in experimental group and control group was not obvious;The expression of vascular endothelial growth factor in experimental group was lower,p < 0.05,with statistical significance.Conclusion1.The protein chip detects cytokines and screens out four differentially expressed cytokines: pulmonary activation regulated chemokine,insulin-like growth factor binding protein 3,vascular endothelial growth factor,and thrombopoietin,of which insulin-like growth factor binding protein 3 is down-regulated and the rest are up-regulated.In the future,the sample can be enlarged,and the biological information analysis can be carried out in combination with the relevant gene database.2.ELISA further verified the expression of four different factors,and statisticalanalysis showed that the expression of vascular endothelial growth factor was significantly lower than that of the control group,suggesting that vascular endothelial growth factor plays an important role in the pathogenesis of autoimmune thyroid disease,speculated that vascular endothelial growth factor through a variety of ways involved in the development of autoimmune thyroid disease,May be a potential biomarker and predictor of disease.