IL-6/IL-6受体mRNA反义寡核苷酸抑制骨吸收的实验研究

Inhibitory effects of IL-6/IL-6 receptor mRNA antisense oligonucleotides on the bone resorption in vitro

目的 探讨 IL- 6及其受体反义寡核苷酸在体外对骨吸收的影响及意义。方法 分离培养新生小鼠颅盖骨器官 ,分别用 IL- 6m RNA反义寡核苷酸 (ASON- 1 )、IL- 6受体 m RNA反义寡核苷酸 (ASON- 2 )、无义寡核苷酸 (NSO)、IL- 6或 IL- 6联合 ASON- 1及 ASON- 2作用培养颅盖骨 ,未加药组为对照。培养 48h后 ,取培养上清液测定钙含量 ,将实验组钙含量与对照组钙含量比值作骨吸收指数。结果 ASON- 1和 ASON- 2均可轻度降低培养颅盖骨的骨吸收指数。二者在浓度为 5μmol/ L时对骨吸收指数的抑制百分率分别为 1 0 .8%和 8.6% ,且 ASON- 1的作用有一定的剂量依赖关系 ;IL- 6作用颅盖骨使骨吸收指数呈剂量依赖性升高 ;当 IL- 6(40 ng/ ml)分别与 5μmol/ L ASON- 1或 5μmol/ L ASON- 2共同作用颅盖骨时 ,IL - 6对骨吸收的刺激性作用均受到抑制 ,其抑制百分率分别为 1 9.3%和 58.0 % ,ASON- 2对 IL- 6的抑制作用明显强于 ASON- 1 ;NSO对骨吸收指数无影响。结论 ASON- 1和 ASON- 2均能轻度抑制正常的颅盖骨骨吸收 ;对 IL- 6刺激导致的颅盖骨骨吸收 ,ASON- 2的抑制作用明显强于ASON- 1。

Objective To investigate the effects and functional significance of IL-6 /IL-6 receptor mRNA antisense oligonucleotides on the bone resorption in vitro.Methods The calvaria of newborn mice was cultured and treated for 48 hours with IL-6 mRNA antisense oligonucleotide (ASON-1), IL-6 receptor mRNA antisense oligonucleotide (ASON-2), nosense oligonucleotide (NSO), IL-6, IL-6 plus ASON-1 or IL-6 plus ASON-2, respectively, and the subjects without above agents as controls group.The calcium concentration in culture medium was detamined by atomic absorption spectrum apparatus.The calcium concentration of treated groups versus that of the control group was used as the bone absorption index.Results The absorption index was slightly decreased when the calvaria was treated with ASON-1 or ASON-2, and the index was bated 10.8% and 8.6% by ASON-1 and ASON-2 with 5 μmol/L respectively. Moreover the inhibitory effect of ASON-1 was in a dose-dependent manner.The absorption index was increased in dose-dependent manner when the calvaria was exposed to IL-6.The stimulating effect of IL-6 ( 40 ng/mL) could be suppressd 19.3% and 58% by ASON-1 and ASON-2 with 5 μmol/L respectively.The absorption index was not varied after the treatment with NSO .Conclusions Both of ASON-1 and ASON-2 can slightly inhibit bone resorption index of normal cultured calvaria and the inhibitory effects of ASON-2 on calvaria bone resorption stimulated by IL-6 were significantly stronger than that of ASON-1.