唑来膦酸对大鼠成骨细胞护骨素及肿瘤坏死因子αmRNA表达的影响

背景:唑来膦酸属于第3代双瞵酸盐类药物,在临床上被广泛应用于治疗骨吸收增加类疾病,其对破骨细胞的作用及影响已取得了共识,而对于成骨细胞的影响尚有争议。目的:观察唑来膦酸对大鼠成骨细胞增殖、分化和护骨素及肿瘤坏死因子αmRNA表达的影响。方法:体外培养新生24h内SD大鼠颅盖骨来源的成骨细胞,用10-5～10-9mol/L唑来膦酸进行干预,分别于干预后第3,5,7天采用MTT法来测吸光度值,以检测唑来膦酸对大鼠成骨细胞增殖的影响;硝基苯基质动力学法和RT-PCR在干预后72h,测量细胞碱性磷酸酶活性和护骨素及肿瘤坏死因子αmRNA的表达。结果与结论:较高浓度(10-5mol/L)的唑来膦酸明显抑制成骨细胞增殖,10-7～10-9mol/L唑来膦酸不影响成骨细胞的分化。10-5～10-9mol/L唑来膦酸能使成骨细胞护骨素mRNA表达增加,肿瘤坏死因子αmRNA表达下降。说明唑来膦酸在低浓度时不影响成骨细胞的增殖及分化,其可通过调节成骨细胞护骨素、肿瘤坏死因子αmRNA的表达,来发挥其抗骨吸收的作用。

Effect of zoledronic acid on osteoprotegerin and tumor necrosis factor alpha mRNA expression in rats

BACKGROUND:Zoledronic acid is a new generation of diphosphate,which is the third generation of osteoporosis drugs.It is widely used to treat diseases related to increase of bone absorption in clinic.The effect of zoledronic acid on osteoclast has reached consensus,but its effect on osteoblast remains in dispute.OBJECTIVE:To observe the effect of zoledronic acid on the proliferation,differentiation and the mRNA expression of osteoprotegerin (OPG) and tumor necrosis factor α (TNF-α) of rat osteoblasts.METHODS:The osteoblasts isolated from the calvaria of neonatal SD rats were cultured with different concentrations of zoledronic acid.The final concentration of zoledronic acid in the experimental group ranged from 10-5-10-9mol/L.At 3,5,and 7 days after zoledronic acid intervention,MTT colorimetric assay was used to measure the absorbance to detect the effect of zoledronic acid on the proliferation of osteoblasts.p-nitrophenyl phosphate approach method was used to detect the activities of alkaline phosphatase,while RT-PCR was used to detect OPG and TNF-α mRNA expression.RESULTS AND CONCLUSION:Zoledronic acid at the concentration of 10-5mol/L could inhibit the proliferation of osteoblasts,but it had no effect at concentration of 10-7-10-9mol/L.Zoledronic acid at concentration of 10-5-10-9mol/L could up-regulate the OPG mRNA expression,but down-regulate TNF-α mRNA expression.Zoledronic acid at low concentration cannot influence the proliferation and differentiation of osteoblasts;whereas,it can inhibit bone resorption by regulating the OPG and TNF-α mRNA expression.