摘要
目的本实验通过体外分离培养兔破骨细胞，观察不同浓度雌激素对兔破骨细胞基质金属蛋白酶MMPmRNA表达的影响。方法体外分离培养出生24h内的新西兰兔破骨细胞，用含有不同浓度17β-雌二醇（0、10^-5～10^-13mol·L^-1）的M199培养液分别作用于破骨细胞，观察不同浓度雌激素及相同浓度雌激素不同时间对破骨细胞活性的影响，采用半定量逆转录-聚合酶链反应（RT-PCR）方法观察兔破骨细胞MMP-9 mRNA、MMP-8 mRNA表达状况。结果不同浓度17β-雌二醇对破骨细胞的活性有不同程度的抑制，同时对MMP-9 mRNA的表达有明显抑制作用，以10^-5、10^-6、10^-7mol·L^-1（P〈0．05）最为显著。所有破骨细胞均未表达MMP-8mRNA。结论不同浓度的雌激素呈时间和剂量依赖性地抑制破骨细胞的活性，对兔破骨细胞基质金属蛋白酶表达的调控作用随其浓度变化而不同。
Objective To observe the effect of estrogen on the expression of matrix metalloproteinases （MMPs） mRNA in rabbit osteoclasts （OCs）. Methods OCs were obtained from the long bone of newborn New Zealand rabbit directly and cultivated in M-199 medium supplemented with 15% fetal bovine serum. Cells were incubated in absence or presence of various concentrations of estrogen （0, 10^-5 ～ 10^-13 mol· L^-1 ）, or treated with estrogen for different period according to the experimental design. Total cellular RNA was isolated with Trizol. Semiquantitative reverse transcription PCR method was used to determine the expression level of MMP-9 mRNA and MMP-8 mRNA. Results Estrogen dose and time-dependently inhibited the activities of osteoclasts. Treatment with estrogen at different concentrations down regulated the expression of MMP-9 mRNA level, especially when the concentrations of estrogen were 10^-5 mol·L^-1 , 10^-6 mol·L^-1 and 10^-7 mol·L^-1 （ P 〈 0.05）. There was no expression of MMP-8 mRNA in osteoclasts despite three repeated experiments. Conclusion Estrogen inhibits osteoclasts and then suppresses the bone resorption activities by decreasing the MMP-9 expression of OCs at mRNA level.
出处
《中国骨质疏松杂志》
CAS
CSCD
2007年第5期301-307,共7页
Chinese Journal of Osteoporosis
基金
Medical research foundation for person with ability of Zhongda Hospital ,Southeast University (2003YJ18)
