川芎嗪对椎间盘退变大鼠模型髓核细胞的保护作用研究

Protective effect of tetramethylpyrazine on nucleus pulposus cells in rat model of intervertebral disc degeneration

目的探讨川芎嗪对椎间盘退变中髓核细胞(NP)凋亡和炎性反应的影响及其机制。方法选取成年SD大鼠54只,随机分为假手术组、对照组、川芎嗪组,每组18只。比较3组盘源性疼痛改善情况及肿瘤坏死因子-α(TNF-α)、白细胞介素-1(IL-1)、微管相关蛋白1轻链3(LC3)-Ⅱ/LC3-Ⅰ和Beclin-1表达情况,并在术后14 d时观察自噬小体形成情况,同时检测NP凋亡水平。结果对照组和川芎嗪组各时间点机械回缩阈值低于假手术组(P<0.05)。川芎嗪组在应用川芎嗪治疗后,疼痛情况较对照组获得明显改善(P<0.01)。对照组和川芎嗪组术后7、14 d时TNF-α和IL-1阳性细胞数量高于假手术组(P<0.01)。川芎嗪组术后7、14 d时TNF-α和IL-1阳性细胞数量低于对照组(P<0.05)。对照组和川芎嗪组术后7、14 d时LC3-Ⅱ/LC3-Ⅰ和Beclin-1表达水平高于假手术组(P<0.01)。川芎嗪组术后7、14 d时LC3-Ⅱ/LC3-Ⅰ和Beclin-1表达水平低于对照组(P<0.05)。川芎嗪组术后14 d时自噬小体形成数量明显低于对照组(P<0.05)。对照组和川芎嗪组术后14 d时NP凋亡水平高于假手术组(P<0.01)。川芎嗪组术后14 d时NP凋亡水平低于对照组(P<0.01)。结论川芎嗪可通过抑制椎间盘退变时炎症细胞因子IL-1和TNF-α表达水平来改善疼痛症状,同时通过有效抑制NP自噬性凋亡的发生来抑制椎间盘退变进展。

Objective To investigate the effect of tetramethylpyrazine on apoptosis and inflammatory reaction of nucleus pulposus(NP)in intervertebral disc degeneration and its mechanism.Methods A total of 54 adult SD rats were randomly divided into sham operation group,control group and ligustrazine group with 18 rats in each group.The improvement of discogenic pain and the expression of tumor necrosis factor-α(TNF-α),interleukin-1(IL-1),microtubule associated protein 1 light chain 3(LC3)-Ⅱ/LC3-Ⅰand Beclin-1 were compared among the three groups.The autophagosome formation and NP apoptosis at 14 days after operation were observed.Results The mechanical retraction thresholds of the control group and the ligustrazine group at each time point were lower than that of the sham operation group(P<0.05).After treatment with ligus-trazine,the pain in the ligustrazine group was significantly improved compared with the control group(P<0.01).The numbers of TNF-αand IL-1 positive cells in the control group and the ligustrazine group at 7 and 14 days after operation were higher than those in the sham operation group(P<0.01).The numbers of TNF-αand IL-1 positive cells in the ligustrazine group at 7 and 14 days after operation were lower than those in the control group(P<0.05).The expression levels of LC3-Ⅱ/LC3-Ⅰand Beclin-1 in the control group and the li-gustrazine group at 7 and 14 days after operation were higher than those in the sham operation group(P<0.01).The expression levels of LC3-Ⅱ/LC3-Ⅰand Beclin-1 in the ligustrazine group at 7 and 14 days after operation were lower than those in the control group(P<0.05).The number of autophagosomes in the ligus-trazine group at 14 days after operation was significantly lower than that in the control group(P<0.05).The levels of NP apoptosis in the control group and the ligustrazine group at 14 days after operation were higher than those in the sham operation group(P<0.01).The level of NP apoptosis in the ligustrazine group at 14 days after operation was lower than that in the control group(P<0.01).Conclusion Ligustrazine can inhibit inflammatory cytokines IL-1 and TNF-αduring intervertebral disc degeneration expression level to improve pain symptoms,and inhibit the progression of intervertebral disc degeneration by effectively inhibiting the oc-currence of NP autophagic apoptosis.