Effects of tumor necrosis factor-α on glucose uptake in human granulosa cells under high androgen conditions

Objective(s): Hyperandrogenism is a key pathological characteristic of polycystic ovary syndrome (PCOS). Tumor necrosis factor α (TNF-α) is both an adipokine and a chronic inflammatory factor, which has been proven to be involved in the pathologic process of PCOS. This study aimed to determine how TNF-α affects glucose uptake in human granulosa cells in the presence of high testosterone concentration. Materials and Methods: KGN cell line was treated with testosterone and TNF-α alone or co-culture combination for ۲۴ hr, or starved for ۲۴ hr. Quantitative real-time polymerase chain reaction (qPCR) and western blot were performed to measure glucose transporter type ۴ (GLUT۴) message RNA (mRNA) and protein expression in treated KGN cells. Glucose uptake and GLUT۴ expression were detected by immunofluorescence (IF). Furthermore, western blot was performed to measure the contents in the nuclear factor kappa-B (NF-κB) pathway. Meantime, upon addition of TNF-α receptor II (TNFRII) inhibitor or Inhibitor of nuclear factor kappa-B kinase subunit beta (IKKβ) antagonist to block the TNFRII-IKKβ-NF-κB signaling pathway, the glucose uptake in KGN cells and GLUT۴ translocation to cytomembrane were detected by IF, and related proteins in TNFRII-IKKβ-NF-κB were detected by western blot.Results: The glucose uptake in Testosterone + TNF-α group was lowered significantly, and Total GLUT۴ mRNA and proteins were reduced significantly. GLUT۴ translocation to cytomembrane was tarnished visibly; concurrently, the phosphorylated proteins in the TNFRII-IKKβ-NF-κB signaling pathway were enhanced significantly. Furthermore, upon addition of TNFRII inhibitor or IKKβ inhibitor to block the TNFRII-IKKβ-NF-κB signaling pathway, the glucose uptake of treated granulosa cells was improved. Conclusion: TNFRII and IKKβ antagonists may improve glucose uptake in granulosa cells induced by TNF-α by blocking the TNFRII-IKKβ-NF-κB signaling pathway under high androgen conditions.

کلیدواژه ها:

glucose uptake ، Granulosa cells ، NF-kappa B signaling - pathway ، Obesity ، Polycystic ovary syndrome ، TNF-α