通经定晕丸阻断颈椎病大鼠NF-κB激活MCP-1/CCR2通路及TNF-α及IL-1β、IL-6表达的研究

目的:研究通经定晕丸阻断颈椎病大鼠核因子-κB(NF-κB)激活单核细胞趋化蛋白-1(MCP-1)/趋化因子受体-2(CCR2)通路及肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、白介素-6(IL-6)表达的影响。方法:用手术的方法制作颈椎病大鼠模型，用通经定晕丸(5.0和10.0 g/kg)和颈复康冲剂(0.84 g/kg)灌胃给药，每天1次，连续4周，每天观察实验大鼠一般状况。4周后检测，腹主动脉取血，进行血液黏度，血液中TNF-α、IL-1β和IL-6含量分析；取颈椎椎间盘进行组织形态学分析；实时定量PCR检测颈椎椎间盘组织中MCP-1和CCR2的mRNA表达，Western blot检测椎间盘组织中NF-κBp65蛋白表达。结果: 通经定晕丸(5.0和10.0 g/kg)和颈复康冲剂(0.84 g/kg)可显著改善颈椎病模型大鼠的一般状况，降低血液低切黏度值、中切黏度值、高切黏度值，促炎因子TNF-α、IL-1β和IL-6含量及椎间盘组织中MCP-1、CCR2和NF-κBp65的mRNA和蛋白表达(P<0.05或P<0.01)，减轻促炎因子对椎间盘造成的损伤，降低颈椎间盘高度和退变程度，降低纤维环裂隙、髓核、细胞数量、软骨厚度，其中以通经定晕丸10.0 g/kg剂量组作用效果更为显著。结论:通经定晕丸对颈椎病模型大鼠具有较好的治疗的作用，其作用机制与降低血液黏度，抑制MCP-1/CCR2信号通路激活、NF-κB活化及TNF-α、IL-1β和IL-6的生成相关。

Study of Tongjing dingyunwan blocking NF-κB activation on the MCP-1/CCR2 signaling pathway and expressions of TNF-α, IL-1β, IL-6 in the cervical spondylosis rats

AIM: To investigate Tongjing dingyunwan blocking nuclear factor-κB (NF-κB) activation on the MCP-1/CCR2 signaling pathway and expressions of TNF-α, IL-1β, IL-6 in the cervical spondylosis rats. METHODS: The cervical spondylosis model was established through operative method. Therapeutic groups were given Tongjing dingyunwan (5.0 g/kg and 10.0 g/kg) and and Jingfukang granule (0.84 g/kg), once a day for four weeks, the general condition of rat was observed. After oral administration of Tongjing dingyunwan for four weeks, blood viscosity, serum TNF-α, IL-1β, and IL-6 were measured; Cervical intervertebral disc morphology was analyzed by HE staining; Real-time PCR was applied to detect the mRNA expressions of MCP-1 and CCR2, protein expression of NF-κBp65 in intervertebral disk tissue were detected by western blotting. RESULTS: Tongjing dingyunwan (5.0 g/kg and 10.0 g/kg) and Jingfukang granule (0.84 g/kg) might significantly improve the general condition of the cervical spondylosis rats, decrease blood low shear viscosity, shear viscosity, viscosity value, serum TNF-α, IL-1β, IL-6 contents and the mRNA and protein expressions of MCP-1, CCR2 and NF-κBp65 in intervertebral disk tissue (P<0.05, P<0.01, respectively), alleviate the proinflammatory cytokine on intervertebral disc injury, reduce the height and degeneration of the cervical intervertebral disc, reduce the fracture of the fibrous ring, the nucleus pulposus, the number of cells and the thickness of the cartilage, especially Tongjing Dingyunwan 10.0 g/kg group. CONCLUSION: Tongjing dingyunwan has a better therapeutic effect on cervical spondylosis rats. Its mechanism is mainly related to reducing blood viscosity, inhibiting the MCP 1/CCR2 signaling pathway, activation of NF-κB, and productions of TNF-α, IL-1β, IL-6.