Study The Role Of MiR-143in Thyroid Associated Ophthalmopathy And The Intravenous Glucocorticoid Therapy

Purpose:The aim of the present study was to explore the mechanisim of miR-143regulating the TSHR expression and HA secretion of orbital fibroblasts in thyroid associated ophthalmopathy pathogenesis. And evaluate adrenal cortex function and efficacy of high-dose glucocorticoid treatment in patients with thyroid associated ophthalmopathy.Methods:Primary cultured TAO and control fibroblasts, immunofluorescence identification. Use Real-time PCR method to detect expression of miRNA-143in patients and control fibroblasts, to verify the accuracy of microarray results. Synthetic antisense oligonucleotide (anti-miR-143) and simulated miR-143(miR-143mimics) to knockdown and raise the levels of miRNA-143in OF cells, detect transfected cells TSHRmRNA and protein expression changes by the method of Real-Time PCR and Western blotting. the miRNA-143target gene was measured by dual luciferase reporter gene assay. at the same time we investigate the HA secretion of fibroblasts by the method of ELISA after transfection.48patients with active, severe TAO patients using a stratified random method into the intermittent treatment group (24cases) and maintenance treatment group (24cases). Two groups of patients, respectively, before the impact of treatment, and24weeks after the end of treatment, every4weeks for efficacy and adverse effects assessment, and to detect the level of serum cortisol (F), adrenocorticotropic hormone (ACTH) and24-hour urinary cortisol to analyze the efficacy and impact of the adrenal cortex function.Results:1.Compared with normal orbital tissue, miR-143gene in TAO patients OF was expressed at low levels, in line with the results of the gene chip.2.miRNA-143 overexpressed fibroblasts express lower TSHR mRNA and TSHR protein levels; contrary, anti-miR-143transfected fibroblasts express elevated TSHR mRNA and protein levels, P <0.05. Dualluciferase reporter gene method confirmed the direct effect of miR-143in the3’UTR of TSHR mRNA.3.decreased expression of miR-143in fibroblasts promotes the secretion of HA.4.of intermittent treatment group, to maintain the efficacy of the treatment group (83.3%,87.5%, invalid or exacerbation rates were16.7%,12.5%, F=0.68; recurrence rates were20%,19.0%, F=0.94) The incidence of adverse reactions (29.2%, respectively,50%, F=0.14) was no significant difference. The adrenocortical function of patients with maintenance treatment was significantly suppressed (P<0.05) and returned to normal in24weeks.Conclusion:Decreased miR-143gene expression in orbital fibroblasts in TAO patients, thus raised the level of TSHR expression and promote secretion of HA in fibroblasts, which was involved in TAO occurrence and development. High dose methylprednisolone intermittent pulse therapy in thyroid associated ophthalmopathy was effective, but maintenance treatment significantly inhibited the adrenal.