氧化槐果碱对腰椎间盘突出模型大鼠的影响研究

Effects of sophoridine oxide in rats with lumbar intervertebral disc herniation

目的观察氧化槐果碱对腰椎间盘突出模型大鼠炎性反应的影响,并初步分析其与核因子-κB(NF-κB)信号通路的关系。方法通过自体髓核移植法构建腰椎间盘突出模型,假手术组仅暴露髓核,不放置自体髓核。将建模成功的SD大鼠随机分为模型组、实验组和对照组,每组16只;实验组腹腔注射氧化槐果碱50 mg·kg-1;对照组腹腔注射吲哚美辛7. 5 mg·kg-1,模型组和假手术组腹腔注射等体积0. 9%NaCl溶液,每日1次,连续干预14 d。用酶联免疫吸附试验(ELISA)测定造模前及造模后第1,3周各组髓核组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、前列腺素E2(PGE-2)含量。以蛋白质印迹(WB)法检测造模后第3周各组大鼠髓核组织中NF-κB p65、磷酸化细胞外信号调节蛋白激酶(p-ERKs)、p-c-Jun氨基末端激酶(p-JNK)蛋白表达情况。结果造模后3周,假手术组、模型组、实验组和阳性对照组大鼠髓核组织中TNF-α分别为(36. 12±4. 75),(165. 35±10. 57),(85. 42±6. 48),(105. 34±8. 45) pg·mg-1,IL-1β分别为(69. 45±1. 58),(205. 69±10. 48),(98. 45±6. 34),(136. 58±4. 51) pg·mg-1,PGE-2分别为(55. 26±3. 48),(126. 58±4. 96),(75. 46±5. 98),(98. 45±6. 75) pg·mg-1,差异均有统计学意义(均P <0. 05)。假手术组、模型组、实验组和对照组大鼠髓核组织中NF-κB p65蛋白相对表达量分别为0. 23±0. 02,1. 34±0. 19,0. 52±0. 14,0. 89±0. 32,p-ERKs蛋白相对表达量为0. 41±0. 05,1. 06±0. 21,0. 56±0. 07,0. 79±0. 13,p-JNK蛋白相对表达量为0. 19±0. 05,1. 32±0. 14,0. 45±0. 09,0. 68±0. 05,差异均有统计学意义(均P <0. 05)。结论氧化槐果碱可有效抑制腰椎间盘突出大鼠炎性反应,缓解椎间盘退变,与抑制NF-κB信号通路及p-ERKs、p-JNK蛋白表达有关。

Objective To investigate the effect of sophoridine oxide on inflammatory response in rats with lumbar disc herniation and to preliminarily analyze its relationship with nuclear factor-kappa B(NF-κB)signaling pathway.Methods The model of lumbar intervertebral disc herniation was established by autologous nucleus pulposus transplantation,only nucleus pulposus was exposed in rats of sham operation group,but no nucleus pulposus autologous was placed.The successfully modeled SD rats were randomly divided into model group,test group and control group,with 16 rats in each group.Test group was intraperitoneally injected with 50 mg·kg-1 sophocarpine oxide;control group was intraperitoneally injected with 7.5 mg·kg-1 indomethacin,model group and sham operation group were intraperitoneally injected with 0.9%NaCl solution of equal volume once a day for 14 d.Enzyme-linked immunosorbent assay(ELISA)was used to determine the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),prostaglandin E2(PGE-2)in nucleus pulposus tissues of each group before modeling and the 1,3 weeks after modeling.The expression of NF-κB p65,phosphorylated extracellular signal-regulated protein kinases(p-ERKs)and p-c-Jun-N-terminal kinase(p-JNK)in nucleus pulposus tissues of rats in each group were detected by Western blot(WB)at the 3 weeks after modeling.Results At 3 week after modeling,the TNF-αwere(36.12±4.75),(165.35±10.57),(85.42±6.48),(105.34±8.45)pg·mg-1,IL-1βwere(69.45±1.58),(205.69±10.48),(98.45±6.34),(136.58±4.51)pg·mg-1,PGE-2 were(55.26±3.48),(126.58±4.96),(75.46±5.98),(98.45±6.75)pg·mg-1,all with significant differece(all P<0.05).The relative expression of NF-κB p65 protein in nucleus pulposus of sham operation group,model group,test group and control group were 0.23±0.02,1.34±0.19,0.52±0.14,0.89±0.32,the relative expression of p-ERKs were 0.41±0.05,1.06±0.21,0.56±0.07,0.79±0.13,the relative expression of p-JNK were0.19±0.05,1.32±0.14,0.45±0.09,0.68±0.05,all with significant differece(all P<0.05).Conclusion Sophoridine oxide can effectively inhibit the inflammatory response and alleviate the degeneration of lumbar intervertebral disc in rats,which is related to the inhibition of NF-κB signaling pathway and the expression of p-ERKs and p-JNK protein.