王少峰,孔祥东,沙 勇,凡 军,高 辉,纪亲龙.紫草素抑制破骨细胞分化及改善去卵巢诱导的小鼠骨质疏松[J].,2017,17(24):4645-4649 |
紫草素抑制破骨细胞分化及改善去卵巢诱导的小鼠骨质疏松 |
Shikonin Suppresses Osteoclastogenesis in Vitro and Ameliorates Ovariectomized-induced Osteoporosis in Mouse Model |
投稿时间:2016-10-12 修订日期:2016-11-10 |
DOI:10.13241/j.cnki.pmb.2017.24.010 |
中文关键词: 紫草素 破骨细胞分化 骨质疏松 核激活因子κB 受体 |
英文关键词: Shikonin Osteoclastogenesis Osteoporosis RANKL |
基金项目:上海市金山区卫生系统“优秀青年人才“培养计划(JSYQ201620) 上海市金山区医疗卫生学科建设基金项目(JSZK2015B06) |
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中文摘要: |
摘要 目的:研究紫草素对破骨细胞体外分化的影响,并探讨其对去卵巢(ovariectomized,OVX)诱导的骨质疏松模型小鼠的骨保护作用。方法:体外细胞生物学实验,采用CCK-8法检测不同浓度紫草素对C57BL/6J小鼠骨髓源性单核巨噬细胞的毒性;采用 RANKL和M-CSF诱导单核巨噬细胞破骨分化模型,给予不同浓度的紫草素干预后,经TRAP染色对破骨细胞进行形态学观察,并通过Real-Time PCR技术检测破骨细胞特异性基因TRAP、c-Fos和NFATc1的表达。动物体内实验,随机将15只小鼠平均分为假手术组、OVX组、治疗组。造模成功后治疗组给予紫草素干预,假手术组和OVX组以等体积生理盐水处理。连续处理30天后取胫骨,用MicroCT扫描重建观察胫骨近端骨丢失状况。结果:①高于250 nmol/L的紫草素显著抑制小鼠单核巨噬细胞生长(P<0.01)。②不同浓度的紫草素干预能显著抑制体外破骨细胞形成(P<0.01)。③不同浓度的紫草素干预能显著抑制TRAP,c-Fos和NFATc1 等参与破骨细胞分化的重要基因表达(P<0.01)。④紫草素干预能显著改善去卵巢诱导的骨质疏松模型小鼠的骨丢失(P<0.05)。结论:紫草素能在体外抑制破骨细胞分化并在体内改善去卵巢诱导的小鼠骨质疏松。 |
英文摘要: |
ABSTRACT Objective: To investigate the effects of shikonin on osteoclastogenesis in vitro and amelioration of bone loss in ovariectomized -induced osteoporosis in mouse model. Methods: The optimal concentration of shikonin treating were evaluated in vitro depending on its effect on the viability of C57BL/6J mouse bone-marrow-derived macrophages by CCK-8 method. To establish the osteoclastogenesis cell model, macrophages were cultured with RANKL and M-CSF treatment, and TRAP staining was used to observe the generation of osteoclasts after treating with different concentration of shikonin solution. Expressions of osteoclast marker genes, including TRAP, c-Fos and NFATc1were detected with real- time PCR. Fifthteen mice were randomly allocated into sham operation group, ovariectomized model group and shikonin treatment group. After the modeling, mice in treatment group were received the intraperitoneal injection of shikonin, while the other two groups treated with normal saline. After thirty days treatments, all animals’ tibias were dissected for micro-CT analysis. Results: ①The macrophages viability was significantly inhibited when the concentration of shikonin was higher than 250 nmol/L(P<0.01). ②The osteoclastogenesis was significantly suppressed by differernt dose of shikonin(P<0.01). ③ The expression of the osteoclastic marker genes (TRAP, c-Fos and NFATc1) were suppressed by addition of shikonin comparing to control group(P<0.01). ④ Shikonin effectively prevented ovariectomy-induced bone loss(P<0.05). Conclusion: Shikonin suppresses osteoclastogenesis in vitro and ameliorates ovariectomized-induced osteoporosis in mouse model. |
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