SIRT2 Mediated Microtubule Acetylation in Osteogenic Differentiation.

Xin Ru Zhou, Can Zhang, Chen Rong Xu, Xin Er Tan, Qian Qian Han, Xi Yang, Tian Yu Sun, Long Quan Shao, Jia Liu
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Abstract

Objective: To assess the role of microtubule acetylation in the transportation of amorphous calcium phosphate (ACP)-containing vesicles that mediate the osteogenic differentiation process of rat bone mesenchymal stem cells (BMSCs).

Methods: Rat BMSCs were cultured and transfected with sirtuin 2 (SIRT2) overexpression plasmids for an in vitro model. The microtubule acetylation-related protein levels were detected by western blots. The microtubule acetylation and the secretion rate of extracellular ACPcontaining vesicles were observed with immunofluorescence and live cell fluorescence imaging. The secretion of ACP was observed by transmission electron microscopy. The mineralised nodule formation was stained with Alizarin Red S staining and observed by microscopy.

Results: Microtubule acetylation was increased during osteogenic differentiation of BMSCs, and microtubule transport efficiency was enhanced. Mechanically, microtubule acetylation is the key reason for the increased transportation rate of ACP-containing vesicles and enhanced osteogenic differentiation, as both were blocked after SIRT2-mediated microtubule acetylation inhibition.

Conclusion: Microtubule acetylation mainly promotes the transportation and secretion of ACP vesicles, and ultimately promotes the osteogenic differentiation process.

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SIRT2介导的微管乙酰化在成骨分化中的作用
目的:探讨微管乙酰化在含无定形磷酸钙(ACP)囊泡转运介导大鼠骨间充质干细胞(BMSCs)成骨分化过程中的作用。方法:培养大鼠骨髓间充质细胞,转染sirtuin 2 (SIRT2)过表达质粒建立体外模型。western blots检测微管乙酰化相关蛋白水平。免疫荧光和活细胞荧光成像观察微管乙酰化及胞外含acp囊泡分泌率。透射电镜观察ACP分泌情况。用茜素红S染色和显微镜观察矿化结节的形成。结果:骨髓间充质干细胞成骨分化过程中微管乙酰化程度增加,微管转运效率提高。机械上,微管乙酰化是acp囊泡运输速率增加和成骨分化增强的关键原因,因为在sirt2介导的微管乙酰化抑制后,两者都被阻断。结论:微管乙酰化主要促进ACP囊泡的转运和分泌,最终促进成骨分化过程。
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