IL-6受体mRNA反义寡核苷酸抑制骨吸收的体外研究

Inhibitory effect of IL-6 receptor mRNA antisense oligonucleotide on mouse bone resorption

目的 :探讨IL 6RmRNA特异反义寡核苷酸 (ASON)在体外对骨吸收的影响及意义。方法 :采用器官培养法培养新生的小鼠颅盖骨 ,并分别用ASON、无义寡核苷酸 (NSO)、IL 6或ASON与IL 6联合处理颅盖骨 ,未加药组为对照。培养一定时间后 ,测定培养上清液的钙含量 ,将实验组钙含量与对照组钙含量比值作骨吸收指数。结果 :作用 4 8h后 ,不同剂量的ASON均可轻度降低骨吸收指数 ,但无剂量依赖关系 ,ASON(5 μmol/L)对正常培养的颅盖骨的骨吸收的抑制率为 8.6 % ;用IL 6培养颅盖骨使骨吸收指数呈剂量依赖关系升高 ,ASON(5 .0 μmol/L)可能明显抑制由IL 6 (4 0μg/L)引起的高骨吸收指数 ,其抑制率为 5 8.0 % ;NSO对骨吸收指数无影响。ASON(5 .0 μmol/L)分别作用 1 2、2 4、4 8h的骨吸收指数则随时间延长而降低。结论 :ASON阻断IL 6R表达后明显抑制IL 6刺激所致的新生小鼠颅盖骨骨吸收 。

Objective:To study the effect of interleukin 6 receptor(IL 6R)mRNA antisense oligonucleotide(ASON)on bone resorption.Methods:Calvaria of newborn mice were cultured,and then exposed to ASON,nonsense oligonucleotide(NSO),IL 6 and IL 6 plus ASON.Calcium concentration of supernatant was determined by atomic absorption spectrophotometry.Bone absorption index(BAI)was defined as the ratio of calcium level in experimental group to that in control group.Results:ASON treatment led to the mild decrease of BAI.but NSO had no obvious influence on BAI.IL 6 treatment resulted in dose dependent increase of BAI,Treated with ASON at the concentration of 5 μmol/L,the inhibition rates of bone absorption for calvarium culture with or without IL 6 were 58% and 8.6% respectively;furthermore,BAI decrease was in a time dependent manner.Conclusion:IL 6R mRNA ASON is via blocking IL 6R expression to inhibit significantly bone absorption of IL 6 stimulated calvarium culture,but its inhibition for bone absorption of normal calvarium culture is mild.