Isobavachin promotes neurogenesis of mouse embryonic stem cells in vitro by regulating miR-124

Abstract

Objective To investigate the effect of isobavachin (IBA) on neurogenesis of mouse J1 embryonic stem cells and explore the mechanism in light of microRNA-124 (miR-124) regulation. Methods The expression of miR-124, octamer transcription factor-4 (OCT4) and neurofilament-H (NFH) were detected by qRT-PCR in J1 cells after treatment with IBA or transfection with miR-124 inhibitor. Targetscan online prediction, dual-luciferase assay and Western blotting were used for verifying the targeting of miR-124 to Zinc finger and BTB domain containing 18 (ZBTB18). The expression levels of OCT4 and NFH were detected with immunofluorescence assay in J1 cells transfected with pcDNA-ZBTB18 or with miR-124 mimics along with ZBTB18 shRNA. Results IBA significantly inhibited the expression of OCT4 and promoted the expression of miR-124 and NFH, and inhibition of miR-124 obviously enhanced OCT4 expression in J1 cells. The rsults of Targetscan online prediction, dual-luciferase assay and Western blotting indicated that miR-124 regulated the expression of ZBTB18 protein. Overexpression of ZBTB18 significantly increased the expression of OCT4 and decreased the expression of NFH in IBA-treated J1 cells; ZBTB18 knockdown in the cells attenuated the regulatory effects of miR-124 on the expression of OCT4 and NFH in IBA-treated J1 cells. Conclusion IBA promotes the expression of miR-124 in J1 cells, and the latter regulates the expression of OCT4 and NFH proteins by targeting ZBTB18, which can be a possible mechanism by which IBA promotes neurogenesis of mouse embryonic stem cells in vitro.

Keywords

• isobavachin
• microrna-124
• zinc finger and btb domain containing 18
• mouse embryonic stem cells
• neurons
• cell differentiation