针刺对穴“后溪-环跳”对腰椎间盘突出症大鼠HMGB1/TLR4/NF-κB信号通路的影响

Effect of Needling Paired Points‘SI3-GB30’on HMGB1/TLR4/NF-κB Signaling Pathway in LDH Rats

目的:观察针刺对穴“后溪-环跳”对LDH大鼠HMGB1/TLR4/NF-κB信号通路及通路相关炎症因子的影响,探讨对穴治疗LDH的潜在机制。方法:将大鼠随机分成假手术组、模型组、对穴组与普通针刺组(即普针组)4组,每组10只。根据自体髓核移植法造模,造模3 d后,对普针组、对穴组行针刺干预,普针组选取双侧“肾俞”“大肠俞”“委中”穴;对穴组取双侧“后溪穴”“环跳穴”。两组均干预1次/d, 30 min/次,共28 d。于造模前1 d、造模3 d后与干预28 d后测定大鼠痛阈值,HE染色观察大鼠L5脊神经干组织病理学改变;ELISA测定血清TLR4、NF-κB、IL-1β、TNF-α与IL-8等含量;Western Blot及免疫荧光技术(IF)检测HMGB1蛋白在脊神经干中的表达。结果:针刺干预后,普针组、对穴组大鼠的痛阈均显著提高;与模型组比较,普针组与对穴组脊神经干中HMGB1蛋白表达及血清中TLR4、NF-κB、IL-1β、TNF-α与IL-8等含量均明显下降,差异具有统计学意义(P<0.05)。结论:针刺能有效缓解LDH大鼠的痛觉敏感,改善脊神经干组织炎症情况,对穴“后溪-环跳”优于普通组穴“肾俞、大肠俞、委中”,其机制可能是通过抑制HMGB1/TLR4/NF-κB信号转导及下调IL-1β、TNF-α和IL-8等炎症因子水平实现的。

Objective:To observe the effect of needling paired points Houxi(SI3)-Huantiao(GB30)on the HMGB1/TLR4/NF-κB signaling pathway and pathway-related inflammatory factors in rats with lumbar disc herniation(LDH),and to explore the potential mechanism of the paired points in treating LDH.Methods:The rats were randomly divided into the sham operation group,the model group,the paired points group and the common acupuncture group,with 10 rats in each group.The model was established according to the autologous nucleus pulposus transplantation method.After 3 days of modeling,the common acupuncture group was treated with needling bilateral Shenshu(BL23),Dachangshu(BL25)and Weizhong(BL40);whereas the paired points group was treated with needling bilateral SI3 and GB30.The intervention was once per day,30 min per time for 28 days in the groups.The pain threshold was measured 1 day before modeling,3 days after modeling,and 28 days after intervention;the histopathological changes of rat L5 spinal nerve trunk were observed by HE staining;the serum contents of TLR4,NF-κB,IL-1β,TNF-αand IL-8 were measured by ELISA;the expression of HMGB1 protein in the spinal nerve trunk was detected by Western Blot and immunofluorescence(IF).Results:The pain threshold was significantly increased in the common acupuncture group and the paired points group after acupuncture intervention.Compared with those in the model group,the expression of HMGB1 protein in the spinal nerve trunk and the serum contents of TLR4,NF-κB,IL-1β,TNF-αand IL-8 were significantly decreased in the common acupuncture group and the paired points group(P<0.05).Conclusion:Acupuncture can effectively relieve the pain sensitivity of LDH rats and improve the inflammation of spinal nerve trunk tissues.Needling SI3-GB30 is better than needling BL23,BL25 and BL40,which mechanism may be related to inhibiting HMGB1/TLR4/NF-κB signaling transduction and down-regulating the levels of inflammatory factors of IL-1β,TNF-αand IL-8.