柚皮苷对破骨细胞分化的影响(1)
[摘要]目的：探讨骨碎补单体成分柚皮苷对小鼠单核细胞RAW264.7诱导分化为破骨细胞的影响。方法：通过100μg·L-1核因子κB受体活化因子配基(RANKL)诱导RAW264.7细胞株分化为成熟破骨细胞，经TRAP特异性染色和骨吸收陷窝对破骨细胞进行鉴定。采用MTT法筛选抑制破骨细胞最强的浓度。在诱导过程中，采用筛选后含柚皮苷的培养基，诱导5d后，通过TRAP阳性细胞计数和骨吸收面积分析来观察柚皮苷对破骨细胞的形成和骨吸收功能情况;流式细胞术检测柚皮苷对破骨细胞增殖的影响，RT-PCR检测柚皮苷对破骨细胞分化过程中核因子κB受体活化因子(RANK)、抗酒石酸酸性磷酸酶(TRAP)、基质金属蛋白酶9(MMP-9)、活化T细胞核因子1(NFATc1)与C-fosmRNA表达的影响。结果：采用100μg·L-1的RANKL可成功诱导成熟的、有功能的破骨细胞。柚皮苷可以抑制破骨细胞的分化和骨吸收功能;抑制破骨细胞的增殖活性;柚皮苷可明显下调破骨细胞分化过程中的RANK，TRAP，MMP-9，NFATc1mRNA的表达，上调C-fosmRNA表达。结论：柚皮苷可抑制破骨细胞分化、增殖和骨吸收功能，其机制可能是通过抑制破骨细胞分化过程中特异性基因表达实现的。
, 百拇医药
[关键词]柚皮苷;破骨细胞;骨质疏松;分化;RAW264.7;MMP-9
Effect of naringin on osteoclast differentiation
LI Feng-bo1， SUN Xiao-lei2， MA Jian-xiong2， ZHANG Yang2， ZHAO Bin2， LI Yan-jun1， MA Xin-long1，2*
(1. Tianjin Medical University General Hospital， Tianjin 300052， China;
2.Institute of Orthopedics in Traditional Chinese and Western Medicine， Tianjin Hospital， Tianjin 300050， China)
, 百拇医药
[Abstract]Objective： To discuss the effect of Drynariae Rhizoma′s naringin on osteoclasts induced by mouse monocyte RAW264.7. Method： RAW264.7 cells were induced by 100 μg·L-1 nuclear factor-κB receptor activator ligand (RANKL) and became mature osteoclasts， which were identified through TRAP specific staining and bone resorption. MTT method was sued to screen and inhibit and the highest concentration of osteoclasts. After being cultured with the screened medium containing naringin for 5 days， positive TRAP cell counting and bone absorption area analysis were adopted to observe the effect of naringin on the formation of osteoclast sells and the bone absorption function. The osteoclast proliferation was measured by flow cytometry. The effects of RANK， TRAP， MMP-9， NFATc1 and C-fos mRNA expressions on nuclear factor-κB were detected by RT-PCR. Result： Naringin could inhibit osteoclast differentiation， bone absorption function and proliferation activity of osteoclasts， significantly down-regulate RANK， TRAP， MMP-9 and NFATc1 mRNA expressions in the osteoclast differentiation process， and up-regulate the C-fos mRNA expression. Conclusion： Naringin could inhibit osteoclast differentiation， proliferation and bone absorption function. Its mechanism may be achieved by inhibiting the specific gene expression during the osteoclast differentiation process.
, http://www.100md.com
[Key words]naringin; osteoclast; osteoporosis; differentiation; RAW264.7; MMP-9
doi：10.4268/cjcmm20150227
骨质疏松(osteoporosis，OP)是以骨量减少，骨微结构破坏，骨脆性增加，易发生骨折为特征的全身性骨病。随着人口老龄化的进程，骨质疏松症逐年呈上升趋势，已成为老年人与绝经后女性的常见病和多发病[1]。根据中医“肾藏精，主骨生髓”的理论，临床上常采用补肾强骨的中药来治疗骨质疏松症。骨碎补为中医临床上常用的补肾壮骨药，其主要有效成分为柚皮苷[2]。许多研究已经证实柚皮苷可以促进成骨细胞的分化、增殖与成骨活性[3]。为进一步探讨柚皮苷是否可以抑制骨吸收活性，本研究在体外观察了柚皮苷对破骨细胞的分化与骨吸收的影响以及其作用机制。, 百拇医药(李风波 孙晓雷 马剑雄 张杨 赵斌 李艳军 马信龙)
, 百拇医药
[关键词]柚皮苷;破骨细胞;骨质疏松;分化;RAW264.7;MMP-9
Effect of naringin on osteoclast differentiation
LI Feng-bo1， SUN Xiao-lei2， MA Jian-xiong2， ZHANG Yang2， ZHAO Bin2， LI Yan-jun1， MA Xin-long1，2*
(1. Tianjin Medical University General Hospital， Tianjin 300052， China;
2.Institute of Orthopedics in Traditional Chinese and Western Medicine， Tianjin Hospital， Tianjin 300050， China)
, 百拇医药
[Abstract]Objective： To discuss the effect of Drynariae Rhizoma′s naringin on osteoclasts induced by mouse monocyte RAW264.7. Method： RAW264.7 cells were induced by 100 μg·L-1 nuclear factor-κB receptor activator ligand (RANKL) and became mature osteoclasts， which were identified through TRAP specific staining and bone resorption. MTT method was sued to screen and inhibit and the highest concentration of osteoclasts. After being cultured with the screened medium containing naringin for 5 days， positive TRAP cell counting and bone absorption area analysis were adopted to observe the effect of naringin on the formation of osteoclast sells and the bone absorption function. The osteoclast proliferation was measured by flow cytometry. The effects of RANK， TRAP， MMP-9， NFATc1 and C-fos mRNA expressions on nuclear factor-κB were detected by RT-PCR. Result： Naringin could inhibit osteoclast differentiation， bone absorption function and proliferation activity of osteoclasts， significantly down-regulate RANK， TRAP， MMP-9 and NFATc1 mRNA expressions in the osteoclast differentiation process， and up-regulate the C-fos mRNA expression. Conclusion： Naringin could inhibit osteoclast differentiation， proliferation and bone absorption function. Its mechanism may be achieved by inhibiting the specific gene expression during the osteoclast differentiation process.
, http://www.100md.com
[Key words]naringin; osteoclast; osteoporosis; differentiation; RAW264.7; MMP-9
doi：10.4268/cjcmm20150227
骨质疏松(osteoporosis，OP)是以骨量减少，骨微结构破坏，骨脆性增加，易发生骨折为特征的全身性骨病。随着人口老龄化的进程，骨质疏松症逐年呈上升趋势，已成为老年人与绝经后女性的常见病和多发病[1]。根据中医“肾藏精，主骨生髓”的理论，临床上常采用补肾强骨的中药来治疗骨质疏松症。骨碎补为中医临床上常用的补肾壮骨药，其主要有效成分为柚皮苷[2]。许多研究已经证实柚皮苷可以促进成骨细胞的分化、增殖与成骨活性[3]。为进一步探讨柚皮苷是否可以抑制骨吸收活性，本研究在体外观察了柚皮苷对破骨细胞的分化与骨吸收的影响以及其作用机制。, 百拇医药(李风波 孙晓雷 马剑雄 张杨 赵斌 李艳军 马信龙)