过表达VCAM-1对小鼠间充质干细胞成脂分化的影响与作用机制

发布时间：2018-05-27 13:38

本文选题：血管细胞粘附分子 + 基因转染　；参考：《中国实验血液学杂志》2015年03期

【摘要】：目的:探讨基因修饰过表达VCAM-1对小鼠间充质干细胞(MSC)成脂分化能力的影响与作用机制。方法:将稳定过表达VCAM-1的MSC细胞(MIGR1-VCAM-1)和转入空载体的MSC细胞(MIGR1)分别向脂肪细胞诱导分化,以原位油红O染色和real-time PCR检测成脂分化能力与相关关键转录因子C/EBPα和PPARγ的表达;Western blot检测相关信号通路P38、ERK和JNK通路的活化。利用信号通路抑制剂处理MSC,观察其成脂分化能力的变化。结果:无论是自分化组还是诱导分化组,过表达VCAM-1的MIGR1-VCAM-1/MSC与对照组MIGR1/MSC相比,脂滴变大,脂肪细胞数量显著增加(P0.01);同时在mRNA水平调控成脂分化的关键转录因子C/EBPα和PPARγ表达显著上调;Western blot检测相关信号通路,结果表明JNK通路在VCAM-1调控成脂分化中明显下调,P38及ERK通路则显著上调;加入通路抑制剂后,JNK通路抑制可显著上调MIGR1-VCAM-1/MSC成脂分化能力,脂肪数量明显增加(P0.01),且相关转录因子C/EBPα和PPARγ的mRNA表达也显著上升;而抑制P38及ERK通路则使MIGR1-VCAM-1/MSC的C/EBPα和PPARγ的mRNA表达水平下调,脂滴及脂肪细胞数则更小、更少。结论:过表达VCAM-1可促进小鼠MSC成脂分化,VCAM-1可能通过抑制JNK信号通路,活化P38及ERK通路促进小鼠MSC成脂分化能力。
[Abstract]:Aim: to investigate the effect and mechanism of gene modified VCAM-1 on adipogenic differentiation of mouse mesenchymal stem cells (MSCs). Methods: MSC cells stably expressing VCAM-1 and MSC cells transfected with empty vector were induced to differentiate into adipocytes, respectively. The ability of adipogenic differentiation and the expression of C/EBP 伪 and PPAR 纬 were detected by in situ oil red O staining and real-time PCR. Western blot was used to detect the activation of P38 ERK and JNK pathways. MSCI was treated with signal pathway inhibitor to observe its ability of adipogenic differentiation. Results: whether in self-differentiation group or induced differentiation group, the lipid droplets of MIGR1-VCAM-1/MSC with overexpression of VCAM-1 were larger than that of control group. At the same time, the expression of C/EBP 伪 and PPAR 纬, the key transcription factors regulating adipogenic differentiation at mRNA level, was upregulated significantly, and the signal transduction pathway was detected by Western blot. The results showed that the JNK pathway down-regulated the P38 and ERK pathway in the regulation of adipogenic differentiation by VCAM-1, and the inhibition of the MIGR1-VCAM-1/MSC pathway induced by the addition of a pathway inhibitor could significantly up-regulate the ability of MIGR1-VCAM-1/MSC to adipogenic differentiation. The expression of mRNA of C/EBP 伪 and PPAR 纬 was increased significantly, while the mRNA expression of C/EBP 伪 and PPAR 纬 of MIGR1-VCAM-1/MSC was down-regulated by inhibition of P38 and ERK pathway, and the number of lipid droplets and adipocytes was smaller and less. Conclusion: overexpression of VCAM-1 can promote the adipogenic differentiation of MSC in mice. VCAM-1 may promote the adipogenic differentiation of MSC by inhibiting the JNK signaling pathway and activating the P38 and ERK pathways.
【作者单位】：天津医科大学总医院儿科;军事医学科学院基础医学研究所细胞生物学研究室;滨州医学院附属医院甲状腺和乳腺外科;河北北方学院研究生部;
【基金】：国家重点基础研究计划(973项目2010CB833600) 国家自然科学基金面上项目(31070996,31171084) 天津市自然科学基金项目(12JCYBJC17100)
【分类号】：R329.28

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