The Role Of The AKT Signaling Pathway In The Pathogenesis Of Hashimoto’s Thyroiditis

Objective:This study focuses on the role of AKT signaling pathway affecting glycolysis in the pathogenesis of Hashimoto’s thyroiditis,and provides new clinical ideas for disease treatment.Methods:All patients who attended the outpatient clinic of the Department of Endocrinology of the Second Affiliated Hospital of Dalian Medical University from 2022/3 to 2022/12 were selected,including 30 HT patients with normal thyroid function and 10 healthy controls.Experiments are conducted in two phases.In the first stage,10 patients with Hashimoto’s thyroiditis were taken as the HT group,10 healthy controls were also selected as the HC group,and in the second stage,20 patients with Hashimoto’s thyroiditis were selected and divided into activation group(Tcc group)and activation followed by AKT inhibitor group(Tcc+AKT inhibitor group),10 patients in each group,and the HT group in the first stage was taken as the inactivation group(UT group).Peripheral blood was collected from the patients,and PBMC were isolated by Ficoll density gradient centrifugation,CD4+ T cells were isolated by human CD4+ T cell isolation kit.The glycolytic capacity of CD4+ T cells was assessed by measuring the concentration of lactate in the CD4+ T cell medium with the Lactate Test Kit,the concentration of glucose in the CD4+ T cell medium with the Glucose Assay Kit and the expression of glucose transporter protein 1(GLUT1)on the surface of CD4+ T cells by flow cytometry,the ratio of CD4+ T cell subpopulations(Treg cells and Th17 cells)and the expression of total AKT and phosphorylated AKT(P-AKT)on the surface of CD4+ T cells by flow cytometry,and the cell proliferation assay by the CCK-8 Assay kit.Results:1.In the first stage,the percentage of CD4+CD25+Treg cells in the total number of CD4+T cells was significantly lower in the HT group compared to the HC group(7.10 ± 1.21)vs(10.02 ± 1.89),the percentage of CD4+IL17+Th17 cells in the total number of CD4+T cells was significantly higher in the HC group(16.34 ± 3.82)vs(11.65 ± 1.89),the expression of GLUT1 on the surface of CD4+T cells was significantly higher in the HT group compared with the HC group(178.89 ± 29.76)vs(150.89 ± 24.09),the level of lactate in CD4+T cell medium was significantly higher in the HT group than in the HC group(3.07 ± 0.46)vs(0.95 ± 0.31),the level of glucose in CD4+T cell medium was significantly lower than in the HC group(5.07 ± 0.46)vs(6.19 ± 0.62),AKT expression on the surface of CD4+T cells was significantly higher in the HT group compared with the HC group(294.80 ± 19.16)vs(245.7 ± 13.04),and the difference was statistically significant(P<0.05).2.In the second stage,the proportion of CD4+CD25+Treg cells was significantly higher in the Tcc group compared to the UT group(59.39 ± 6.48)vs(7.10 ± 1.20),the proportion of CD4+IL17+Th17 cells was significantly higher in the Tcc group compared to the UT group(58.04 ± 7.12)vs(16.34 ± 3.82),the expression of GLUT1 on the surface of CD4+T cells was significantly higher in the Tcc group compared to the UT group(316.30 ± 46.11)vs(178.89 ± 29.76)and the level of lactate in CD4+T cell medium was significantly higher in the Tcc group(4.48 ± 0.26)vs(3.07 ± 0.46)compared to the UT group,with statistically significant differences(P<0.05).The proportion of CD4+CD25+Treg cells was significantly higher in the Tcc+AKT inhibitor group compared to the Tcc group(80.19 ± 7.86)vs(59.39 ± 6.48),and the proportion of CD4+IL17+Th17 cells was significantly lower in the Tcc+AKT inhibitor group compared to the Tcc group(45.59 ± 8.74)vs(58.04 ± 7.12).The expression of GLUT1 on the surface of CD4+T cells was significantly lower in the Tcc+AKT inhibitor group compared to the Tcc group(239.2 ± 56.07)vs(316.30 ± 46.11),and the concentration of lactate in CD4+T cell medium was significantly lower in the Tcc+AKT inhibitor group compared to the Tcc group(3.75 ± 0.14)vs(4.48 ± 0.26),with statistically significant differences(P<0.05).Conclusion:1.In patients with Hashimoto’s thyroiditis,the ratio of Th17/Treg cells is imbalanced,there is abnormal expression of AKT signaling pathway in peripheral blood CD4+ T cells,and the glycolysis pathway is abnormally activated.2.After adding AKT inhibitors to CD4+T cells in peripheral blood of HT patients,the expression and activation of AKT were significantly inhibited,the glycolytic pathway of CD4+T cells was inhibited,and the proportion imbalance of Th17/Treg cells was restored,suggesting that the AKT signaling pathway may participate in the occurrence of HT by regulating the imbalance of glycolysis affecting Th17/Treg cells.