Roles of matricellular CCN2 deposited by osteocytes in osteoclastogenesis and osteoblast differentiation
Roles of matricellular CCN2 deposited by osteocytes in osteoclastogenesis and osteoblast differentiation
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骨细胞沉积的基质细胞CCN2在破骨细胞生成和成骨细胞分化中的作用
DOI：
10.1038/s41598-019-47285-3
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发表时间：
2019
影响因子：
4.6
通讯作者：
Takigawa M
中科院分区：
文献类型：
--
作者：
Nishida T;Kubota S;Yokoi H;Mukoyama M;Takigawa M
在这项研究中，我们研究了沉积在骨基质中的CCN2（细胞通讯网络因子2）（以前称为结缔组织生长因子）对破骨细胞生成和成骨细胞分化的影响。为了模拟骨基质环境，将骨细胞 MLO-Y4 细胞嵌入含有重组 CCN2 (rCCN2) 的胶原凝胶中，并将小鼠巨噬细胞样 RAW264.7 细胞接种在凝胶上并用 NF-κB 配体受体激活剂 (RANKL) 处理。用 rCCN2 处理的 RAW264.7 和 MLO-Y4 细胞组合中的 NFATc1 和组织蛋白酶 K (CTSK) 产量比没有 rCCN2 的组合增加更多。接下来，我们从野生型和他莫昔芬诱导的 Ccn2 缺陷 (KO) 小鼠中分离出富含骨细胞的细胞群和破骨细胞祖细胞，并进行了类似的分析。无论破骨细胞祖细胞的来源如何，注射他莫昔芬后 6 个月，富含 KO 骨细胞的群体中 NFATc1 和 CTSK 的产量均下降。有趣的是，注射后 1 年，KO 骨细胞中 CTSK 的产量相当增加。最后，成骨细胞MC3T3-E1和MLO-Y4细胞在含有rCCN2的骨基质中的组合揭示了成骨细胞标记基因的上调。这些发现表明，由骨细胞提供的 CCN2 调节破骨细胞生成和成骨细胞分化。
In this study, we investigated the effect of CCN2 (cellular communication network factor 2), previously termed connective tissue growth factor, deposited in bone matrix on osteoclastogenesis and osteoblast differentiation. To mimic the bone matrix environment, osteocytic MLO-Y4 cells had been embedded in collagen-gel with recombinant CCN2 (rCCN2), and mouse macrophage-like RAW264.7 cells were inoculated on the gel and treated with receptor activator of NF-κB ligand (RANKL). NFATc1 and cathepsin K (CTSK) productions were more increased in the combination of RAW264.7 and MLO-Y4 cells treated with rCCN2 than the combination without rCCN2. Next, we isolated an osteocyte-enriched population of cells and osteoclast progenitor cells from wild type and tamoxifen-inducibleCcn2-deficient (KO) mice and performed similar analysis. NFATc1 and CTSK productions were decreased in the KO osteocyte-enriched population at 6 months after the tamoxifen injection, regardless of the origin of the osteoclast progenitor cells. Interestingly, CTSK production was rather increased in KO osteocytes at 1 year after the injection. Finally, the combination of osteoblastic MC3T3-E1 and MLO-Y4 cells in rCCN2-containing bone matrix revealed the up-regulation of osteoblastic marker genes. These findings suggest that CCN2 supplied by osteocytes regulates both osteoclastogenesis and osteoblast differentiation.