Resveratrol attenuates senescence of adipose-derived mesenchymal stem cells and restores their paracrine effects on promoting insulin secretion of INS-1 cells through Pim-1

摘要

OBJECTIVE: The paracrine effects of mesenchymal stem cells (MSCs) were weakened during aging. This study explored whether resveratrol can attenuate senescence of adipose-derived MSCs (ADMSCs) and whether Pim-1 is involved in resveratrol’s effect on paracrine of ADMSCs and insulin secretion of INS-1 cells. MATERIALS AND METHODS: CCK-8 assay and SA-b-gal assay were performed to test the protective effect of resveratrol on senescent models. QRT-PCR and western blot analysis were performed to analyze of senescence- and β-cell associated genes. QRT-PCR and ELISA analysis was performed to test telomere length and activity. Immunofluorescence and ELISA assay were performed to assess the paracrine effects on promoting insulin secretion of INS-1 cells. RESULTS: Resveratrol could protect ADMSCs from H2O2 and D-glucose-induced senescence and also attenuate senescence in long-term cultured ADMSCs in vitro. In addition, resveratrol attenuated H2O2 induced higher expression of senescence-associated genes, including P53, P21, Cyclin D1, IL-6 and MMP1, but increased the expression of Sirt1, a well-known anti-senescence gene. Resveratrol significantly enhanced Pim-1 expression in aging ADMSCs through PI3K/AKT signal pathway. The conditioned medium (CM) of ADMSCs enhanced insulin secretion and expression of the key genes for β-cell function including TFAM, PDX1, GLUT2 and HNF-1α via Pim-1. INS-1 cells with Pim-1 knockdown had decreased insulin secretion. CONCLUSIONS: This study firstly reported that resveratrol has a protective effect on senescence of ADMSCs and can preserve the paracrine effect of the ADMSCs on promoting insulin secretion of INS-1 cells via Pim-1. Therefore, it might be a promising adjuvant agent for future MSCs based therapy.