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目的探讨辛伐他汀与地塞米松对成骨细胞内皮细胞型一氧化氮合酶(endothelialnitricoxidesynthase,eNOS)基因表达及增殖的影响。方法从新生SD大鼠颅盖骨分离培养成骨细胞,随机分为3组,A组加入1×10-7mol/L地塞米松、1×10-7mol/L辛伐他汀和二甲亚砜(DMSO,终浓度为1‰);B组加入1×10-7mol/L地塞米松和等量DMSO;C组仅加等量DMSO作为对照。96h后提取细胞RNA,RT-PCR一步法分析成骨细胞eNOSmRNA表达,硝酸还原酶法测定细胞培养液中NO含量,MTT法测定细胞增殖率。结果3组细胞eNOSmRNA表达比值分别为0.491±0.014、0.421±0.018与0.489±0.014。NO含量分别为(14.37±1.24)μmol/L、(12.94±0.69)μmol/L、(14.14±1.23)μmol/L。细胞增殖率分别为0.3055±0.0178、0.2659±0.0105、0.3044±0.0213。三个指标A组与B组、B组与C组比较差异均有统计学意义(P<0.05),A组与C组比较差异均无统计学意义(P>0.05)。结论地塞米松导致成骨细胞eNOS基因低表达,辛伐他汀能够对抗地塞米松而调控成骨细胞eNOS基因表达,保护成骨细胞增殖。
Objective To investigate the effect of simvastatin and dexamethasone on the gene expression and proliferation of endothelial nitric oxide synthase (eNOS) in osteoblasts. Methods Osteoblasts were isolated and cultured from the calvaria of neonatal SD rats and randomly divided into three groups. Group A received 1 × 10-7mol / L dexamethasone, 1 × 10-7mol / L simvastatin and dimethyl sulfoxide (DMSO, final concentration was 1 ‰). B group was treated with dexamethasone (1 × 10-7 mol / L) and isoflurane (DMSO). After 96h, the cell RNA was extracted. The expression of eNOS mRNA in osteoblasts was analyzed by RT-PCR. The content of NO was determined by nitrate reductase method. The cell proliferation rate was determined by MTT assay. Results The ratio of eNOS mRNA expression in the three groups was 0.491 ± 0.014, 0.421 ± 0.018 and 0.489 ± 0.014, respectively. The levels of NO were (14.37 ± 1.24) μmol / L, (12.94 ± 0.69) μmol / L and (14.14 ± 1.23) μmol / L, respectively. Cell proliferation rates were 0.3055 ± 0.0178, 0.2659 ± 0.0105 and 0.3044 ± 0.0213, respectively. There was significant difference between group A and group B, group B and group C (P <0.05). There was no significant difference between group A and group C (P> 0.05). Conclusion Dexamethasone leads to low expression of eNOS gene in osteoblasts. Simvastatin can modulate the expression of eNOS gene in osteoblasts against dexamethasone and protect osteoblast proliferation.