Int J Stem Cells.  2020 Mar;13(1):142-150. 10.15283/ijsc19073.

IL6 Receptor Facilitates Adipogenesis Differentiation of Human Mesenchymal Stem Cells through Activating P38 Pathway

• Deng W
• Chen H
• Su H
• Wu X
• Xie Z
• Wu Y
• Shen H

• KMID: 2500573
• DOI: http://doi.org/10.15283/ijsc19073

Abstract

Background and Objectives
Mesenchymal stem cells (MSCs) have the multipotent capacity to differentiate into multiple tissue lineages as well as to self-renew, which is the main origin of adipocytes. IL6/IL6R pathway exerts a significant role in tissue regeneration and cell differentiation. Whereas, the underlying mechanism between IL6/IL6R pathway and MSCs adipogenesis differentiation remains elusive.
Methods
MSCs from healthy donors were cultured in adipogenesis differentiation medium for 0∼14 days, during which their adipogenesis differentiation degree was evaluated by Oil Red O staining. The expression of IL6R was detected in MSCs during adipogenesis differentiation. Knockdown and overexpression of IL6R were respectively performed using siRNA and lentivirus to investigate its effect on MSCs adipogenesis differentiation. The adipogenesis marker genes expression and MAPK pathway activation were detected by Western blotting. The role of P38 pathway in the adipogenesis differentiation of MSCs was determined using the specific inhibitor SB203580.
Results
The expression of IL6 and IL6R increased during adipogenesis differentiation in MSCs, which were positively correlated with Oil Red O quantification result. Knockdown and overexpression experiments demonstrated a positive correlation between the expressions of IL6R and MSCs adipogenesis differentiation, accompanied by same trend of P38 phosphorylation. Besides, the specific P38 inhibitor SB203580 markedly inhibited the adipogenesis differentiation potential of MSCs.
Conclusions
This study reveals IL6R facilitates the adiogenesis differentiation of MSCs via activating P38 pathway.

Keyword

Mesenchymal stem cells (MSCs); Adipogenesis differentiation, Interleukin 6 (IL6); Interleukin 6 receptor (IL6R); P38

Figure

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  Fig. 1 Immunophenotype investigation and induced differentiation of MSCs. (A) MSCs were positive for CD29, CD44, CD90 and CD105 and negative for CD14, CD45, and HLA-DR. (B) MSCs was induced to differentiate into osteoblasts, chondrocytes and adipocytes.

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  Fig. 2 Correlation between MSCs adipogenesis differentiation and IL6R. (A) MSCs adipogenic differentiation from day0 to day14 was assessed by Oil red O staining and quantification. (B) IL6R expression was significantly increased from day0 to day14 during induction. (C) IL6R expression was positively correlated with Oil red O staining results in MSCs. (D) IL6 secretion was significantly increased from day0 to day14 during induction. (E) IL6 secretion was positively correlated with Oil red O staining results in MSCs. Data is displayed as the means±SD. * represents p<0.05.

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  Fig. 3 Downregulation of IL6R inhibited adipogenesis differentiation of MSCs (A, B) The ability of adipogenesis differentiation of MSCs decreased after IL6R siRNA knockdown. (C∼E) Decreased expression of MSCs adipogenic differentiation marker genes CEBR-α and PPAR-γ after knockout of IL6R. * represents p<0.05.

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  Fig. 4 Upregulation of IL6R enhanced adipogenesis differentiation of MSCs (A, B) The adipogenesis differentiation of MSCs was enhanced after IL6R overexpression. (C∼E) Enhanced expression of MSCs adipogenic differentiation marker genes CEBR-α and PPAR-γ after IL6R overexpression. * represents p<0.05.

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  Fig. 5 Involvement of P38 phosphorylation in the adipogenesis differentiation-enhancing effect of IL6R. (A, C) After IL6R knockout, the activation of P38 pathway was inhibited and there was no significant change in ERK or JNK. (B, D) After IL6R overexpression, the activation of P38 pathway was enhanced, and ERK as well as JNK showed no significant changes. (E) Oil Red O staining on day 10 of adipogenesis differentiation was significantly reduced by SB203580. * represents p<0.05.

Reference

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