摘要
目的:检测骨碎补有效成分柚皮苷对成骨细胞株MC3T3-E1细胞增殖、分化和矿化的影响。方法:以成骨细胞株MC3T3-E1细胞为体外药效的试验模型(药物组和对照组),通过CCK-8法观察10,1,0.1,0.01,0.001,0.000 1μmol.L-1的柚皮苷溶液对MC3T3-E1细胞增殖能力的影响;通过乳酸脱氢酶(LDH)细胞毒性试验观察以上浓度的柚皮苷溶液对细胞毒性能力的影响。通过骨形成蛋白-2(BMP-2),碱性磷酸酶(ALP)活性测定和骨钙素(OC)含量测定观察柚皮苷对MC3T3-E1细胞分化能力的影响;通过Von kossa钙化染色法观察柚皮苷对MC3T3-E1细胞钙化能力的影响。结果:高浓度的柚皮苷溶液在12 h和24 h时能促进MC3T3-E1细胞的增殖作用。而低浓度的柚皮苷溶液则无此作用。所测的各个组别细胞毒性百分比都较小,且随着时间的推移(12,24,48 h)变化较小。光镜下观察各个时间点细胞的密度和形态也未发生明显的变化。BMP-2细胞免疫化学结果表明,24,48 h时柚皮苷浓度10,1,0.1μmol.L-1包浆内棕色着色比对照组明显。ALP检测结果表明48 h时,1,0.1μmol.L-1的柚皮苷溶液可提高MC3T3细胞的ALP活性(P<0.05)。72 h时,0.1μmol.L-1的柚皮苷溶液可提高MC3T3细胞的ALP活性(P<0.05)。OC检测结果表明12 d时,10,1μmol.L-1柚皮苷溶液作用组与对照组相比OC活性明显提高(P<0.05)。Vonkossa染色钙化面积百分比结果表明3组柚皮苷溶液作用组与对照组相比无明显区别。结论:柚皮苷溶液可以提高成骨细胞株MC3T3-E1增殖能力和分化能力。
Objective： To investigate the effect of naringin on the proliferation, differentiation and matrix mineralization of MC3T3-E1 cells in vitro. Method： MC3T3-E1 cell lines were taken in vitro model. CCk-8 method was used to observe the proliferation of MC3T3 cells. Lactic acid dehydrogenase cytotoxicity （LDH） test was used to observe the cell toxicity. Bone morphogenetic protein- 2 （BMP-2）, alkaline phosphatase （ALP） and osteocalcin （OC） were used to observe the cell differentiation. Van kossa calcification staining method was used to observe the cell calcification. Result： The high dosages of the naringin could promote the proliferation of MC3T3-E1 cells at both 12 h and 24 h. While, low dosages did not show the same capability. LDH test showed that the cytotoxicity percentages in all six naringin treated groups were quite low. BMP-2 eytoimmunochemistry test showed that the three naringin treated group （10, 1,0. 1 umol·L^-1 ） showed higher brown coloration in cytoplasm than the control group at both 24 h and 48 h. 1, 0. 1 umol·L^-1 naringin raised ALP activity of MC3T3-E1 cells at 48 h （P 〈0. 05）. Meanwhile, 0. 1umol·L^-1 naringin increased the ALP activity at 72 h （P 〈 0. 05 ）. 10 and 1umol·L^-1 naringin increased the capability of MC3T3-E1 cell to synthesize osteocalcin during 8^th-12^th dsince adding the medicine （P 〈 0. 05）. Naringin did not show the positive effects on cell calcification. Conclusions： Naringin could promote proliferation and differentiation of MC3T3-E1 cells.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2009年第13期1712-1716,共5页
China Journal of Chinese Materia Medica
基金
浙江省科技厅重点科研项目(2005c23034)
关键词
柚皮苷
成骨细胞
增殖
分化
基质矿化
naringin
osteoblast
proliferation
differentiation
matrix mineralization
