Combined treatment with vitamin K2 and PTH enhanced bone formation in ovariectomized rats and increased differentiation of osteoblast in vitro
Combined treatment with vitamin K2 and PTH enhanced bone formation in ovariectomized rats and increased differentiation of osteoblast in vitro
复制标题
维生素 K2 和 PTH 联合治疗可增强去势大鼠的骨形成并增加体外成骨细胞的分化
DOI：
10.1016/j.cbi.2019.01.012
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发表时间：
2019-02-25
影响因子：
5.1
通讯作者：
Yang, Lei
中科院分区：
文献类型：
--
作者：
Weng, She-Ji;Yan, De-Yi;Yang, Lei
骨质疏松症伴有成骨能力不足。一些证据表明，促进成骨细胞生成的解决方案是骨质疏松性骨缺损修复的重要策略。本研究探讨维生素K2和甲状旁腺素联合应用对骨质疏松大鼠颅骨缺损骨形成的影响及其对成骨细胞的影响。采用SPF级SD大鼠双侧卵巢切除建立骨质疏松模型。随后建立大鼠颅骨缺损模型，将所有骨质疏松大鼠随机分为对照组、维生素K2组(维生素K2，每天30 mg/kg)、PTH组(重组人甲状旁腺素(1~34)，60µg/kg，每周3次)和VK+PTH组(维生素K2 30 mg/kg，每天3次+PTH 60 mg/kg，每周3次)，共8周。体外培养骨髓间充质干细胞，用维生素K2、甲状旁腺素或维生素K2+甲状旁腺素处理。用碱性磷酸酶染色和免疫印迹法观察联合用药对BMSCs的影响。采用血清γ-羧化骨钙素(GLA-OC)、显微CT、组织学和免疫荧光标记技术评价颅骨缺损区的骨形成情况。在本研究中，甲状旁腺激素和维生素K2联合治疗对预防去卵巢大鼠股骨骨丢失有积极作用。联合治疗可提高骨质疏松性颅骨缺损区血清GLA-OC水平，促进骨形成。免疫组织化学结果显示，VK+PTH组OCN和RUNX2的表达均高于对照组。体外实验结果表明，甲状旁腺素和维生素K2联合作用可增加BMSCs碱性磷酸酶、骨形态发生蛋白2和RUNX2的表达。结果提示，维生素K2和甲状旁腺素联合应用可促进成骨细胞分化，对骨质疏松性颅骨缺损区的骨形成有协同作用。
Osteoporosis is accompanied by insufficient osteogenic capacity. Several lines of evidence suggested that solutions to enhance osteoblastogenesis were important strategies for osteoporotic bone defect repair. This study investigated the effect of combined treatment with vitamin K2 and PTH on bone formation in calvarial bone defect in osteoporotic rats and its influence on osteoblast in vitro. Bilateral ovariectomy was used in SPF Sprague Dawley rats to generate an osteoporosis model. Subsequently, a calvarial defect model was established and all osteoporotic rats were randomly assigned to the following groups: control, VK (vitamin K2, 30 mg/kg everyday), PTH (recombinant human PTH (1-34), 60 mu g/kg, three times a week) or VK + PTH (vitamin K2, 30 mg/kg everyday plus PTH, 60 mu g/kg three times a week) for 8 weeks. In vitro, bone marrow-derived stem cells (BMSCs) were cultured and treated with vitamin K2, PTH or vitamin K2 + PTH. ALP staining and western blot were performed to observe the influence of combined treatment on BMSCs. Bone formation within calvarial defect were assessed by serum gamma-carboxylated osteocalcin (Gla-OC), micro-CT, histological and immunofluorescent labeling. In this study, combined treatment of PTH and vitamin K2 showed positive effects on preventing bone loss in femurs in OVX rats. Combined treatment increased serum Gla-OC and promoted bone formation in osteoporotic calvarial bone defects. Immunohistochemistry showed that OCN and RUNX2 were more highly expressed in the VK + PTH group than in the control groups. In vitro studies results suggested that combined treatment with PTH and vitamin K2 increased expression of ALP, BMP2 and RUNX2 in BMSCs. Our data suggested that the combination of vitamin K2 and PTH increased differentiation of osteoblast and had a synergistic effect on bone formation in osteoporotic calvarial bone defect.