【摘要】 目的研究不同浓度的TNF-α及TNF-α抗体对破骨细胞上V-ATP酶表达量的影响。方法体外诱导小鼠RAW264.7细胞分化为破骨细胞,通过抗酒石酸酸性磷酸酶染色检测破骨细胞生成情况。然后将破骨细胞分为对照组、TNF-α干预组及TNF-α抗体干预组,TNF-α干预组、TNF-α抗体干预组分别用低、中、高三种浓度的TNF-α、TNF-α抗体干预48 h。用实时荧光定量聚合酶链反应(real-time PCR)、Western blot检测破骨细胞V-ATP酶的mRNA和蛋白表达水平。结果 TRAP染色检测提示有多核破骨细胞生成。TNF-α处理组V-ATP酶mRNA表达水平显著高于对照组(P<0.001);TNF-α抗体处理组V-ATP酶mRNA表达水平显著低于对照组(P<0.001)。同时,TNF-α处理组V-ATP酶蛋白表达水平显著高于对照组(P<0.05);TNF-α抗体处理组V-ATP酶蛋白表达水平显著低于对照组(P<0.05)。结论 TNF-α可提高破骨细胞V-ATP酶的表达;TNF-α抗体可抑制破骨细胞V-ATP酶的表达。上述提示TNF-α可能通过提高破骨细胞V-ATP酶的表达从而增加破骨细胞的骨吸收作用。更多还原

【Abstract】 Objective To explore the effect of different concentrations of TNF-alpha and TNF-alpha antibody on the V-ATPase expression in osteoclasts. Methods Mouse RAW264. 7 cells were induced into osteoclasts in vitro. The osteoclastogenesis was detected with tartrate resistant acid phosphatase( TRAP) staining. The osteoclasts were divided into control group,TNF-alpha intervention group,and TNF-alpha antibody intervention group. TNF-alpha antibody intervention group was treated with different concentrations of TNF-alpha antibody for 48 h. The mRNA and the protein expression levels of V-ATPase in osteoclasts were examined using real-time PCR and Western blotting analysis. Results The formation multinucleated osteoclasts were proved with TRAP staining. The mRNA expression level of V-ATPase in TNF-alpha treatment group were significantly higher than that in the control group( P < 0. 001). The mRNA expression level of V-ATPase in TNF-alpha antibody treatment group were significantly lower than that in the control group( P < 0. 001). The protein expression levels of V-ATPase in TNF-alpha treatment group were significantly higher than that in the control group( P < 0. 05). The protein expression level of V-ATPase in TNF-alpha antibody treatment group were significantly lower than that in the control group( P < 0. 05). Conclusion TNF-alpha improves the expression of V-ATPase in osteoclasts. TNF-alpha antibody reduces the expression of V-ATPase in osteoclasts. These suggest that TNF-alpha improves osteoclastic bone absorption by increase the expression of V-ATPase.更多还原