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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MMP mRNA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-8 mRNA。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 investigate the effects of different concentrations of estrogen on the expression of matrix metalloproteinase (MMP) mRNA in rabbit osteoclasts by in vitro isolation and culture of rabbit osteoclasts. Methods New Zealand rabbit osteoblasts were isolated and cultured 24h in vitro. M199 culture medium containing different concentrations of 17β-estradiol (0, 10-5-10-13 mol·L-1) Different concentrations of estrogen and the same concentration of estrogen on the activity of osteoclasts were detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) method to observe the expression of MMP-9 mRNA and MMP-8 mRNA in osteoclasts situation. Results Different concentrations of 17β-estradiol inhibited the activity of osteoclasts to a certain extent and inhibited the expression of MMP-9 mRNA at 10-5, 10-6, 10-7 mol·L-1 ( P <0.05) the most significant. None of the osteoclasts expressed MMP-8 mRNA. CONCLUSION: Different concentrations of estrogen can inhibit the activity of osteoclasts in a time and dose-dependent manner. The regulation of matrix metalloproteinase expression in osteoclasts varies with the concentration of MMPs.