miR-141-3p对腰椎间盘突出症大鼠髓核组织核转录因子κB信号通路的影响

Effects of miR-141-3p on the nuclear factor-κB signaling pathway in the nucleus pulposus tissue of rats with lumbar disc herniation

目的探讨miR-141-3p对腰椎间盘突出(LDH)大鼠髓核组织核转录因子κB(NF-κB)信号通路的影响。方法使用自体髓核回植法对20只8周龄雄性SD大鼠建立腰椎间盘突出模型,最终成功建立模型18只;取出模型大鼠髓核组织均分成miR-141-3p mimics转染组、转染miR-141-3p阴性对照组及不进行转染正常对照组;3组大鼠髓核组织培养48 h,比较各组大鼠髓核组织的细胞活力、侵袭及凋亡,比较各组大鼠髓核组织的炎性指标白细胞介素-1(IL-1)、IL-10、肿瘤坏死因子-α(TNF-α)、红细胞沉降率(ESR)、C反应蛋白(CRP)水平、NF-κB信号通路相关因子核因子κB磷酸化65(NF-κB p65)、IκB激酶α(IKK-α)、磷酸化IκB激酶α(p-IKK-α)、IκB激酶α抗体(IKB-α)、磷酸化IκB激酶α抗体(p-IKB-α)及其mRNA表达水平。结果转染组大鼠髓核组织培养48 h时的细胞活力和细胞侵袭个数,NF-κB p65、IKK-α、p-IKK-α、IKB-α、p-IKB-α水平及NF-κB p 65、IKK-α、IKB-αmRNA水平低于阴性对照组和正常对照组,凋亡率高于阴性对照组和正常对照组,差异有统计学意义(P<0.05);3组大鼠髓核组织培养48 h时的IL-1、TNF-α、ESR及CRP水平比较,转染组<阴性对照组<正常对照组,差异有统计学意义(P<0.05);3组大鼠髓核组织培养48 h时的IL-10水平比较,转染组>正常对照组>阴性对照组,差异有统计学意义(P<0.05)。结论miR-141-3p高表达能够对NF-κB信号通路的激活产生抑制作用,进而抑制髓核细胞活力和侵袭,促进细胞凋亡,缓解炎症反应。

Objective To investigate the effect of miR-141-3p on the nuclear factorκB(NF-κB)signaling pathway in the nucleus pulposus tissue of rats with lumbar disc herniation(LDH).Methods Autologous nucleus pulposus replantation was used to establish lumbar disc herniation models on 208-week-old male SD rats,and 18 models were successfully established.The nucleus pulposus tissue was removed and divided into 3 groups(transfected with miR-141-3p mimics transfection group,transfected miR-141-3p negative control group,and no transfection normal control group).Three groups of nucleus pulposus tissues were cultured for 48 hours,and the cell viability,invasion and apoptosis of the nucleus pulposus tissues in each group were compared.Comparing the inflammatory index(IL-1,IL-10,TNF-α,ESR,CRP level,NF-κB p65,IKK-α,p-IKK-α,IKB-α,p-IKB-α)levels of nucleus pulposus tissues,and their mRNA expression levels in each group.Results Cell viability and cell invasion number of nucleus pulposus tissue cultured for 48 hours in transfection group,NF-κB p65,IKK-α,p-IKK-α,IKB-α,p-IKB-αlevels and NF-κB p 65,IKK-α,IKB-α mRNA levels were lower than the negative control group and normal control group,and the apoptosis rate was higher than negative control and normal control group,difference were statistically significant(P<0.05).Comparing levels of IL-1,TNF-α,ESR,CRP levels cultured in the nucleus pulposus tissue for 48 hours of 3 groups,the transfection group<negative control group<normal control group,differences were statistically significant(P<0.05);comparing IL-10 levels of three groups of nucleus pulposus tissue culture at 48h,the transfection group>normal control group>negative control group,differences were statistically significant(P<0.05).Conclusion The high expression of miR-141-3p can inhibit the activation of NF-κB signaling pathway,thereby inhibiting the viability and invasion of nucleus pulposus cells,promoting cell apoptosis and alleviating inflammation.