吴茱萸碱通过调节核因子E2相关因子2/血红素加氧酶-1/核因子-κB信号通路对白细胞介素1β诱导的软骨细胞炎性损伤的影响  

Impact of evodiamine on IL-1β-induced chondrocyte inflammatory injury by regulating the Nrf2/HO-1/NF-κB signaling pathway

作　　者：周凡[1] 高扬[1] 胡艳平[1] 向超 熊和然 周茹 ZHOU Fan;GAO Yang;HU Yanping;XIANG Chao;XIONG Heran;ZHOU Ru(Department of Massage,Wuhan Hospital of Traditional Chinese Medicine,Wuhan,Hubei 430000;Department of Encephalopathy,Wuhan Hospital of Traditional Chinese Medicine,Wuhan,Hubei 430000)

机构地区：[1]湖北省武汉市中医医院推拿科,湖北武汉430000 [2]湖北省武汉市中医医院脑病科,湖北武汉430000

出　　处：《河北中医》2024年第12期2029-2034,共6页Hebei Journal of Traditional Chinese Medicine

基　　金：武汉市医学科研项目(编号:WZ21C09)。

摘　　要：目的观察吴茱萸碱(Evo)对白细胞介素1β(IL-1β)诱导的软骨细胞炎性损伤的影响及对核因子E2相关因子2(Nrf2)/血红素加氧酶-1(HO-1)/核因子κB(NF-κB)信号通路的调节作用。方法将人软骨细胞HCCs分为对照组、模型组、Evo低剂量组、Evo高剂量组、Evo高+ML385组,对照组用DMEM培养基培养,其他组用加入50 ng/mL的IL-1β培养基培养,同时Evo低、高剂量组再加浓度分别为10、30μmol/L的Evo处理,Evo高+ML385组先用2μmol/L的Nrf2抑制剂ML385处理30 min,再用30μmol/L的Evo处理。CCK-8检测细胞增殖活力;流式细胞术检测HCCs细胞凋亡;酶联免疫吸附法检测HCCs细胞上清液中IL-6、IL-18、肿瘤坏死因子α(TNF-α)、丙二醛(MDA)、超氧化物歧化酶(SOD)水平;实时荧光定量聚合酶链式反应(qRT-PCR)法检测HCCs细胞中炎症介质环氧合酶2(COX-2)、诱导型一氧化氮合酶(iNOS)mRNA水平;Western blot法检测HCCs中Nrf2/HO-1/NF-κB通路相关蛋白表达。结果与对照组相比,模型组凋亡率、IL-6、IL-18、TNF-α、MDA水平,COX-2、iNOS mRNA表达水平,以及磷酸化NF-κB抑制蛋白α(p-IκB-α)、细胞质Nrf2、细胞核p65 NF-κB蛋白水平均升高(P<0.05),细胞存活率、SOD活性及HO-1、IκB-α、细胞核Nrf2、细胞质p65 NF-κB蛋白水平均降低(P<0.05)。与模型组相比,Evo低、高剂量组细胞凋亡率、IL-6、IL-18、TNF-α、MDA水平,COX-2、iNOS mRNA表达水平,p-IκB-α、细胞质Nrf2、细胞核p65 NF-κB蛋白水平均降低(P<0.05),细胞存活率、SOD活性及HO-1、IκB-α、细胞核Nrf2、细胞质p65 NF-κB蛋白水平均升高(P<0.05),且Evo高剂量组以上指标与Evo低剂量组组间比较差异也均有统计学意义(P<0.05)。与Evo高剂量组相比,Evo高+ML385组中Nrf2的抑制剂ML385消除了Evo高剂量对上述指标的影响。结论Evo可能通过激活Nrf2和HO-1的表达,抑制下游转录因子NF-κB的表达,改善IL-1β诱导的软骨细胞炎性损伤。Objective To investigate the impact of evodiamine(Evo)on interleukin-1beta(IL-1β)-induced chondrocyte inflammatory injury and its regulation on the nuclear factor erythrocyte 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)/nuclear factor kappa B(NF-κB)signaling pathway.Methods Human chondrocyte cells line(HCCs)were grouped into the control group,model group,low-dose group,high-dose group,and high-dose Evo+Nrf2 inhibitor ML385 group.In addition to HCCs treated with blank control in the control group,those in the remaining groups were induced with 50 ng/mL IL-1β.Treatment of 10μmol/L Evo,30μmol/L Evo,and 30-min induction of 2μmol/L ML385 followed by 30μmol/L Evo treatment was conducted in the latter three groups,respectively.Cell viability and apoptosis were detected by cell counting kit-8(CCK-8)assay and flow cytometry,respectively.Enzyme-linked immunosorbent assay(ELISA)was applied to detect the levels of interleukin-6(IL-6),IL-18,tumor necrosis factor-α(TNF-α),malondialdehyde(MDA),superoxide dismutase(SOD)in the supernatant of HCCs cells.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was applied to detect the mRNA levels of inflammatory mediators cyclooxygenase 2(COX-2)and inducible nitric oxide synthase(iNOS)in HCCs cells.Western blot was applied to detect the protein expressions of Nrf2/HO-1/NF-κB pathway-related proteins in HCCs cells.Results Compared with those of the control group,HCCs in the model group had significantly higher apoptotic rate,supernatant levels of IL-6,IL-18,TNF-αand MDA,mRNA levels of COX-2 and iNOS,protein levels of p-IκB-α,cytoplasmic Nrf2 and nuclear p65 NF-κB,but lower cell viability,SOD activity,and protein levels of HO-1,IκB-α,nuclear Nrf2 and cytoplasmic p65 NF-κB(P<0.05).Compared with those of the model group,HCCs in the low-dose and high-dose Evo groups had significantly lower apoptotic rate,supernatant levels of IL-6,IL-18,TNF-αand MDA,mRNA levels of COX-2 and iNOS,protein levels of p-IκB-α,cytoplasmic Nrf2 and nuclear p65 NF-κB,but hi

关 键 词：骨关节炎 软骨细胞 吴茱萸碱 核因子E2相关因子2 血红素加氧酶-1 核因子-κB 白细胞介素1Β 炎症反应 

分 类 号：R684.3[医药卫生—骨科学] R285.5[医药卫生—外科学]