雷公藤红素抑制磨损颗粒诱导细胞炎症反应的研究

Inhibitory Effects of Tripterine on Wear Particle-Induced Cellular Inflammatory Reaction

目的通过体外细胞培养,探讨雷公藤红素抑制人工关节磨损颗粒诱导的无菌性炎症反应的效果。方法使用真空球磨法制备人工关节磨损颗粒并用无血清培养基配制颗粒悬液。使用RAW264.7巨噬细胞进行传代培养,并将其按不同处理因素随机分成4组:空白对照组(A组)、磨损颗粒组(B组)、磨损颗粒+雷公藤红素组(C组)和雷公藤红素组(D组)。分组处理24 h后行CCK-8毒性检测;通过ELISA检测各组肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β表达情况;RT-PCR检测TNF-α、IL-1β和核因子(NF)-κB的基因表达量;Western Blot在蛋白水平检测NF-κB的表达情况。结果 1mg/L雷公藤红素的细胞毒性作用不明显;磨损颗粒组促炎症因子TNF-α、IL-1β的表达和NF-κB的激活明显高于空白对照组(P<0.05),加入雷公藤红素后以上各指标表达均明显降低(P<0.05)。结论雷公藤红素可在基因和蛋白水平抑制磨损颗粒诱导RAW264.7细胞促炎症因子的表达,并可抑制NF-κB通路的激活。

Objective To investigate the effect of tripterine on wear particle-induced inflammatory reaction by cell culture in vitro. Methods Wear particles from artificial joints were prepared by vacuum ball milling, and made into the par- ticle suspension by using serum-free medium. RAW264.7 cells were cultured, subcultured and divided randomly into four groups according to different treatment factors： blank control group （group A）, wear particle group （group B）, wear particle ＋ tripterine group （group C） and tripterine group （group D）. After 24 hours, the toxicity of tripterine was detected by CCK-8 assay. ELISA was used to detect the expressions of tumor necrosis factor-α （TNF-α） and interleukin-1β（IL-1β）. RT-PCR was used to detect the expressions of TNF-α, IL-1β and nuclear factor-KB （NF-KB） at the gene level. Western blot assay was used to detect the expression of NF- KB at the protein level. Results The results showed that 1 mg/L tripterine was little cytotoxic. The expressions of pro-inflammatory cytokines and the activation of NF-kB were significantly higher in wear particle group than those in blank control group （P 〈 0.05）, which were significantly decreased after the treatment with tripterine （group C, P 〈 0.05）. Conclusion Tripterine can inhibit the wear particle-induced expression and release of pro-inflammatory cytokines at the gene level and protein level and the activation of NF-KB in RAW264.7 cells.