Deng Wen, Chen Huadi, Su Hongjun, Wu Xiaohua, Xie Zhongyu, Wu Yanfeng, Shen Huiyong
Center for Biotherapy, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China.
Organ Transplant Center, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
Int J Stem Cells. 2020 Mar 30;13(1):142-150. doi: 10.15283/ijsc19073.
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.
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.
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.
This study reveals IL6R facilitates the adiogenesis differentiation of MSCs via activating P38 pathway.
间充质干细胞（MSCs）具有多能分化为多种组织谱系以及自我更新的能力，是脂肪细胞的主要来源。IL6/IL6R通路在组织再生和细胞分化中发挥重要作用。然而，IL6/IL6R通路与MSCs成脂分化之间的潜在机制仍不清楚。
将健康供体的MSCs在成脂分化培养基中培养0至14天，在此期间通过油红O染色评估其成脂分化程度。检测MSCs在成脂分化过程中IL6R的表达。分别使用小干扰RNA（siRNA）和慢病毒进行IL6R的敲低和过表达，以研究其对MSCs成脂分化的影响。通过蛋白质免疫印迹法检测成脂标记基因的表达和丝裂原活化蛋白激酶（MAPK）通路的激活情况。使用特异性抑制剂SB203580确定P38通路在MSCs成脂分化中的作用。
MSCs在成脂分化过程中IL6和IL6R的表达增加，与油红O定量结果呈正相关。敲低和过表达实验表明IL6R的表达与MSCs成脂分化呈正相关，同时P38磷酸化呈现相同趋势。此外，特异性P38抑制剂SB203580显著抑制了MSCs的成脂分化潜能。
本研究揭示IL6R通过激活P38通路促进MSCs的成脂分化。
