TSH inhibits eNOS expression in HMEC-1 cells through the TSHR/PI3K/AKT signaling pathway.

OBJECTIVE: To investigate the effects of thyroid-stimulating hormone (TSH) on the expression of endothelial nitric oxide synthase (eNOS) in human microvascular endothelial cells (HMEC-1) and explore the potential mechanism. MATERIALS AND METHODS: Expression of thyroid-stimulating hormone receptor (TSHR) in HMEC-1 cells was determined by immunofluorescence, reverse transcription-polymerase chain reaction (RT-PCR), and Western blotting. Cell proliferation and the production of nitric oxide (NO) and superoxide anion (SA) were measured after TSH treatment. eNOS expression and AKT phosphorylation were detected by Western blotting. RESULTS: TSHR was expressed in HMEC-1 cells. TSH promoted HMEC-1 cell proliferation and SA production, but inhibited NO generation by dose-dependent blocking of mRNA and protein expression of eNOS. Mechanism studies demonstrated that TSH promoted AKT phosphorylation (P<0.05), and that LY294002 inhibited the reduction of eNOS expression by TSH. Moreover, TSH activated the AKT signaling pathway through binding to TSHR on HMEC-1 cells. CONCLUSIONS: TSH inhibits NO production via the TSHR/AKT signaling pathway.