Liu Yamei, Xu Liangliang, Hu Liuchao, Chen Dongfeng, Yu Lijuan, Li Xican, Chen Hongtai, Zhu Junlang, Chen Chen, Luo Yiwen, Wang Bin, Li Gang
School of Basic Medical Science, Guangzhou University of Chinese Medicine, Guangzhou 510006, China.
The Research Center of Basic Integrative Medicine, Guangzhou University of Chinese Medicine, Guangzhou 510006, China.
J Orthop Translat. 2019 Oct 23;22:81-91. doi: 10.1016/j.jot.2019.09.008. eCollection 2020 May.
Mesenchymal stem cells (MSCs) can be easily expanded without losing the ability of multilineage differentiation, including oesteogenic, chondrogenic and adipogenic differentiation. These characters make MSCs a promising cell resource for cartilage defect repair. MSCs could be recruited by inflammatory stimulation, then home to the injury tissues. However, its capacity of homing is extremely limited. Thus, it has become extremely necessary to develop an agent or a method, which can be used to enhance the efficiency of MSCs homing. This study investigates the effect of stearic acid methyl ester (SAME) on MSCs mobilisation and cartilage regeneration.
MSCs were isolated from femurs of Sprague-Dawley (SD) rats. MTT assay was used to detect effect of SAME on viability of MSCs. Transwell assay and wound healing assay were used to detect effect of SAME on migration of MSCs. RNA-seq, quantitative real-time PCR and western blot were performed to analyze the expression of RNAs and proteins. Colony forming assay and flow cytometry were used to evaluate the effect of SAME on MSCs mobilisation in vivo. A rat cartilage defect model was created to evaluate the effect of SAME on cartilage regeneration.
We found that SAME could promote the migration of MSCs. Interestingly, we found SAME significantly increased the expression levels of Vav1 in MSCs. On the other hand, the enhanced migration ability of MSCs induced by SAME was retarded by Vav1 small interfering RNA (siRNA) and Rho-associated protein kinase 2 (ROCK2) inhibitor. In addition, we also checked the effect of SAME on mobilisation of MSCs in vivo. The results showed that SAME increased the number of MSCs in peripheral blood and enhanced the capacity of colony formation. Finally, using a cartilage defect model in rats, we found SAME could improve cartilage repair.
Our study demonstrates that SAME can enhance MSCs migration ability mainly through the Vav1/ROCK2 signaling pathway, which could contribute to the accelerated cartilage regeneration.
These findings provide evidence that SAME could be used as a therapeutic reagent for MSCs mobilisation and cartilage regeneration.
间充质干细胞（MSCs）易于扩增，且不会丧失多向分化能力，包括成骨、成软骨和脂肪生成分化。这些特性使MSCs成为软骨缺损修复中一种有前景的细胞资源。MSCs可被炎症刺激募集，然后归巢至损伤组织。然而，其归巢能力极其有限。因此，开发一种可用于提高MSCs归巢效率的试剂或方法变得极为必要。本研究探讨硬脂酸甲酯（SAME）对MSCs动员和软骨再生的影响。
从Sprague-Dawley（SD）大鼠的股骨中分离出MSCs。采用MTT法检测SAME对MSCs活力的影响。采用Transwell法和伤口愈合试验检测SAME对MSCs迁移的影响。进行RNA测序、定量实时PCR和蛋白质印迹分析RNA和蛋白质的表达。采用集落形成试验和流式细胞术评估SAME对体内MSCs动员的影响。建立大鼠软骨缺损模型以评估SAME对软骨再生的影响。
我们发现SAME可促进MSCs的迁移。有趣的是，我们发现SAME显著增加了MSCs中Vav1的表达水平。另一方面，Vav1小干扰RNA（siRNA）和Rho相关蛋白激酶2（ROCK2）抑制剂可抑制SAME诱导的MSCs迁移能力增强。此外，我们还检测了SAME对体内MSCs动员的影响。结果表明，SAME增加了外周血中MSCs的数量并增强了集落形成能力。最后，使用大鼠软骨缺损模型，我们发现SAME可改善软骨修复。
我们的研究表明，SAME可主要通过Vav1/ROCK2信号通路增强MSCs的迁移能力，这可能有助于加速软骨再生。
这些发现提供了证据，表明SAME可作为一种治疗试剂用于MSCs动员和软骨再生。
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