目的 检测在人成纤维细胞成骨分化过程中肌动蛋白的改变及其介导的线粒体动力学改变。 方法 诱导人成纤维细胞成骨分化，检测肌动蛋白和线粒体动力学的改变。在人成纤维细胞中打断（细胞松弛素D，Cyto D）或者促进（Jasplakinolide，JAS）肌动蛋白的聚合，于处理的第1、4、7 d分别进行免疫荧光和Western blot等实验检测细胞性能及线粒体动力学相关指标。 结果 Cyto D减弱成骨诱导液效果，JAS增强成骨诱导液效果。JAS增强线粒体动力学效果比Cyto D更加显著。 结论 肌动蛋白的聚合促进细胞成骨分化；肌动蛋白的聚合或解聚影响细胞成骨分化过程中线粒体与细胞骨架肌动蛋白的共定位比率。Cyto D和JAS引起的对细胞成骨分化的影响和线粒体动力学的改变，可为研究细胞线粒体以及成骨分化机制提供生物学信息。
Abstract
Objective To explore the affects of actin on osteogenic differentiation of human fibroblast by regulating mitochondrial dynamics. Methods Cytochalasin D (Cyto D) and Jasplakinolide (JAS) were added to be the growth medium and osteogenic differentiation medium respectively, then the changes of cell characters and mitochondrial dynamics were detected by Immunofluorescence staining and western blot experiments in the treatment of 1 d, 4 d and 7 d. Results Cyto D reduced the effect of osteogenic induction media, but JAS increased the effect of it. JAS enhanced significantly the effect of mitochondrial dynamics in fibroblast than Cyto D. Conclusions Actin polymerization motivates the osteogenic differentiation; Actin polymerization and depolymerization affects the colocalization ratio between mitochondrial and actin in osteogenic differentiated cells. The results can provide important biological information for studying the mitochondrial and differentiation mechanism.
关键词
线粒体 /
成骨分化 /
细胞松弛素D /
细胞骨架稳定剂
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参考文献
[1] Suen DF, Norris KL, Youle RJ. Mitochondrial dynamics and apoptosis[J]. Genes Dev, 2008, 22(12): 1577-1590.
[2] Yang C, Svitkina TM. Ultrastructure and dynamics of the actin-myosin II cytoskeleton during mitochondrial fission[J]. Nat Cell Biol, 2019, 21(5): 603-613.
[3] Li M, Zhang C, Yang Y. Effects of mechanical forces on osteogenesis and osteoclastogenesis in human periodontal ligament fibroblasts: a systematic review of in vitro studies[J]. Bone Joint Res, 2019, 8(1): 19-31.
[4] Koerdt S, Siebers J, Bloch W, et al. Role of oxidative and nitrosative stress in autogenous bone grafts to the mandible using guided bone regeneration and a deproteinized bovine bone material[J]. J CranioMaxillofac Surg, 2014, 42(5): 560-567.
[5] Hadjiargyrou M, O'Keefe RJ. The convergence of fracture repair and stem cells: interplay of genes, aging, environmental factors and disease[J]. J Bone Miner Res, 2014, 29(11): 2307-2322.
[6] Li S, Xu S, Roelofs BA, et al. Transient assembly of F-actin on the outer mitochondrial membrane contributes to mitochondrial fission[J]. J Cell Biol, 2015, 208(1): 109-123.
[7] 黄健, 姜小凡, 叶莉斯, 等. 线粒体动力学研究进展[J]. 吉林医药学院学报, 2018, 39(5): 371-373.
[8] Shares BH, Busch M, White N, et al. Active mitochondria support osteogenic differentiation by stimulating beta-catenin acetylation[J]. J Biol Chem, 2018, 293(41): 16019-16027.
[9] Seo JH, Agarwal E, Bryant KG, et al. Syntaphilin ubiquitination regulates mitochondrial dynamics and tumor cell movements[J]. Cancer Res, 2018, 78(15): 4215-4228.
[10] 樊庭宇, 曲戎梅, 冯雁婷, 等. 不同浓度细胞松弛素D对人脂肪源干细胞分化能力的影响[J]. 中国临床解剖学杂志, 2018, 36(1): 45-50.
[11] 邵开峰, 吴泽志, 王伯初, 等. 细胞骨架对肝癌细胞粘附性的影响[J]. 重庆大学学报(自然科学版), 2000, 23(3): 9-11.
[12]Han Y, Kim SJ. Simvastatin-dependent actin cytoskeleton rearrangement regulates differentiation via the extracellular signal-regulated kinase-1/2 and p38 kinase pathways in rabbit articular chondrocytes[J]. Eur J Pharmacol, 2018, 834: 197-205.
[13]Steffen J, Koehler CM. ER-mitochondria contacts: actin dynamics at the ER control mitochondrial fission via calcium release[J]. J Cell Biol, 2018, 217(1): 15-17.
[14]Wu F, Ye H, Lin J, et al. TGF-beta3 reduces apoptosis in ischemia-induced adipose-derived stem cells by enhancing DNA repair[J]. Exp Ther Med, 2018, 15(5): 4400-4408.
基金
国家重点研发计划资助项目（2017YFC1105000）；深圳市医疗卫生三名工程（SZSM201612019）
京ICP备08002354号-3
