腰突颗粒对人髓核细胞核因子κB信号通路的影响

摘要

背景：核因子κB在退变椎间盘中呈高表达，经验方腰突颗粒治疗腰椎间盘突出症临床效果良好，通过核因子κB信号通路试探讨其治疗作用的机制。目的：探讨腰突颗粒含药血清对人髓核细胞核因子κB信号转导通路的影响。方法：将8只新西兰大白兔随机等分为生理盐水组和低、中、高剂量组，分别以生理盐水及10，20，40 mL腰突颗粒水煎剂灌胃7 d。于最后一次灌胃2 h后颈总动脉采血，收集血清，分别配成体积分数10%含药血清的DMEM培养液保存备用，分别干预人髓核细胞24 h。结果与结论：体积分数10%含药血清干预后，与生理盐水组相比，低剂量组、中剂量组、高剂量组人髓核细胞中白细胞介素8、肿瘤坏死因子α、p-P65、P50、IκB-α和IKK-βmRNA相对表达水平均降低，且细胞中p-P65、P50、IκB-α和IKK-β蛋白表达水平降低，同时随着灌服腰突颗粒剂量的增加，白细胞介素8、肿瘤坏死因子α、p-P65、P50、IκB-α和IKK-βmRNA相对表达水平逐渐降低，p-P65、P50、IκB-α和IKK-β蛋白表达水平降低。提示腰突颗粒含药血清能阻断人髓核细胞中核因子κB信号转导通路，进而减缓髓核细胞的退变，起到保护人髓核细胞的作用。%BACKGROUND:Nuclear factor-κB (NF-κB) is highly expressed in the degenerated intervertebral disc. The Herbal Compound formula Yaotu Granules for lumbar disc herniation has achieved good clinical results;however, the underlying mechanisms remain poorly understood. OBJECTIVE:To explore mechanisms underlying the effects of the drug serum of Yaotu Granules on NF-κB signaling pathway. METHODS:Eight New Zealand white rabbits were randomly divided into normal saline group, low-, middle-, and high-dose drug groups. Normal saline, 10, 20, and 40 mL of water decoction of Yaotu Granules were administered intragastrical y in the normal saline, low-, middle-, and high-dose drug groups for 7 days, respectively. The drug serum of Yaotu Granules was prepared using serum samples obtained from the common carotid artery at 2 hours after the last administration. Human nucleus pulposus cel s were maintained in DMEM containing 10%(v/v) drug serum for 24 hours. RESULTS AND CONCLUSION:The expression levels of interleukin (IL)-8, tumor necrosis factor (TNF)-α, p-P65, P50, IκB-α(the NF-κB inhibitor), and IκB kinaseβ(IKK-β) mRNA in human nucleus pulposus cel s in three drug serum groups were significantly decreased compared with the normal saline group;in addition, a similar change in the expression levels of p-P65, P50, IκB-α, and IKK-βprotein between groups were detected. Noticeably, the expression levels of IL-8, TNF-α, p-P65, P50, IκB-α, and IKK-βmRNA, as wel as the expression levels of p-P65, P50, IκB-α, and IKK-βprotein were gradual y decreased with the increased drug dose. Al these results suggest that the drug serum of Yaotu Granules can protect nucleus pulposus cel s and delay their degeneration through blocking NF-κB signaling pathway.