Effect of 1,25(OH)2D3 on type I collagen secretion in adipose-derived mesenchymal stem cells and its mechanism

doi:10.3969/j.issn.2095-4344.0466

Abstract

Abstract:

BACKGROUND: Adipose-derived mesenchymal stem cells (ADMSCs) have been reported to improve wound healing. However, type I collagen secreted by ADMSCs will contribute to scar formation. Therefore, inhibiting type I collagen secretion from ADMSCs will strengthen its clinical application.
OBJECTIVE: To investigate the effect of 1,25(OH)₂D₃ on secretion of type I collagen by ADMSCs and its mechanism.
METHODS: Human ADMSCs were isolated by collagenase digestion, and identified by flow cytometry. ADMSCs at passage 4 were cultured in DMEM/F12 medium containing different concentrations of 1,25(OH)₂D₃ (10^-⁷, 10^-⁸, 10^-⁹, 10^-¹⁰ and 0 mol/L) respectively for 4 days. Then, the concentration of type I collagen in cell supernatant was measured by ELISA. Real-time PCR and western blot were used to detect the expression of Smad3 at mRNA and protein levels and phosphorylated protein Smad3 level in ADMSCs cultured with and without 1,25(OH)₂D₃. To analyze the contribution of Smad3 to the effect of 1,25(OH)₂D₃, Smad3 inhibitor was added to culture medium 30 minutes before adding 1,25(OH)₂D₃, and type I collagen in cell supernatant was detected by ELISA at 4 days after addition of SMAD3 inhibitor.
RESULTS AND CONCLUSION: 1,25(OH)₂D₃ inhibited the secretion of type I collagen by ADMSCs in a dose-dependent manner. The results of real-time PCR and western blot showed that the expression of Smad3 was upregulated by 1,25(OH)₂D₃, and the results of western blot showed that the phosphorylated Smad3 protein level in ADMSCs was significantly increased by 1,25(OH)₂D₃. Moreover, the inhibition of type I collagen secretion by 1,25(OH)₂D₃could be blocked by Smad3 inhibitor. These results indicate that 1,25(OH)₂D₃ can inhibit the secretion of type I collagen from ADMSCs by up-regulating the expression of Smad3.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Adipose Tissue, Mesenchymal Stem Cells, Calcitriol, Collagen Type I, Smad3 Protein, Tissue Engineering

CLC Number:

R394.2

Zhai Min, Hu Xiao-gen, Liu Hong-lin, Xu Shi-qing, Wang Zai, Zhang Wen-jian. Effect of 1,25(OH)₂D₃ on type I collagen secretion in adipose-derived mesenchymal stem cells and its mechanism[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(9): 1370-1375.

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References

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Effect of 1,25(OH)₂D₃ on type I collagen secretion in adipose-derived mesenchymal stem cells and its mechanism

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