Melatonin promotes Schwann cell migration by activating a typical Wnt/β-catenin signaling pathway

doi:10.3969/j.issn.2095-4344.2138

Abstract

Abstract:

BACKGROUND: Melatonin, a neuroendocrine hormone, is mainly secreted by the pineal gland, which can promote the proliferation of Schwann cells and the regeneration of peripheral nerve after injury.

OBJECTIVE: To investigate the role of melatonin in the migratory activity of Schwann cells together with the molecular mechanism involved.

METHODS: The rodent RSC96 Schwann cell line was cultured in vitro followed by identification. The CCK-8 assay was performed to detect the effect of melatonin on Schwann cells proliferation. The Transwell chamber was adopted to observe the efficacy of melatonin with a series of concentrations, which is 0, 100 nmol/L, 1 μmol/L and 10 μmol/L, on the migratory activity of Schwann cells following 20 hours. The expression of melatonin receptor 1 and 2 in Schwann cells was assessed using qRT-PCR and western blot assay. Transwell cell migration system was conducted to determine the influence of melatonin receptor antagonist luzindole or inhibitor IWR-1 of the canonical Wnt/β-catenin signaling on the enhanced migratory activity exerted by melatonin. Western blot assay was performed to investigate the regulatory effect of melatonin on the activity of Wnt/β-catenin signal cascades.

RESULTS AND CONCLUSION: (1) The cultured RSC96 cells showed the typical morphology of Schwann cells, showing a spindle-like or triangular shape with thin and long processes on the two cell poles. The Schwann cells marker S-100 immunofluorescence staining was positive. (2) Compared with the blank control group, 1 μmol/L and 10 μmol/L melatonin could promote Schwann cell migration dramatically compared to the control group, and displayed a chemoattractive effect, with 1 μmol/L showing the most significant effect. (3) Melatonin receptor 1 was the dominating receptor of melatonin expressed in Schwann cells. The Transwell assay results displayed that the number of migratory cells that melatonin induced was not significantly changed with luzindole added, as compared with treatment with melatonin alone, showing no statistical significance. The number of Schwann cells treated with IWR-1 and melatonin was significantly lower than those treated with melatonin (P < 0.05). (4) Results of qRT-PCR and western blot assay showed that compared with the blank control group, the expression levels of P-Lrp6, LEF-1 and nuclear β-catenin protein in Schwann cells of melatonin group were significantly increased (P < 0.05). (5) The expression levels of LEF-1 and nuclear β-catenin protein in Schwann cells of melatonin group and IWR-1 + melatonin group were significantly higher than those of blank control group and IWR-1 group (P < 0.05), indicating that melatonin can antagonize the inhibition of IWR-1 on Wnt/β-catenin signaling pathway activity to some extent. (6) The above data confirm that melatonin can promote Schwann cell migration by activating Wnt/β-catenin signaling pathway.

Key words: melatonin, Schwann cell, factor, pathway, cell migration, peripheral nerve regeneration, protein, experiment

CLC Number:

Lü Jianwei, Ma Jianxiong, Ma Xinlong. Melatonin promotes Schwann cell migration by activating a typical Wnt/β-catenin signaling pathway[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(31): 5030-5037.

Figures/Tables 6

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