TNF-α抑制剂对风湿性膝关节滑膜炎大鼠氧化应激抑制作用及下游信号通路的影响

Effect of TNF-α inhibitor on oxidative stress inhibition and downstream signaling pathway in rats with rheumatic knee synovitis

目的分析肿瘤坏死因子-α(TNF-α)抑制剂对风湿性膝关节滑膜炎大鼠氧化应激抑制作用及下游信号通路的影响。方法取36只SD大鼠,随机分为对照组(正常大鼠)、模型组(风湿性膝关节滑膜炎+生理盐水)、干预组(风湿性膝关节滑膜炎+TNF-α抑制剂),每组12只。经苏木精-伊红染色行病理学检查,检测血清TNF-α、白细胞介素(IL)-1β、IL-6、基质金属蛋白酶9(MMP-9)和C反应蛋白;同时测定血清超氧化物歧化酶(SOD)、谷胱甘肽和丙二醛水平;Western blot检测膝关节滑膜组织TNF-α转换酶(TACE)和核转录因子-κB(NF-κB)蛋白表达,并进行统计学分析。结果病理检查显示模型组、干预组大鼠关节组织出现明显病理变化。相比对照组,模型组和干预组血清TNF-α、IL-1β、IL-6、MMP-9、C反应蛋白及丙二醛水平均明显升高,谷胱甘肽和SOD水平均明显降低(P<0.05);相比模型组,干预组血清TNF-α、IL-1β、IL-6、MMP-9、C反应蛋白、丙二醛水平均明显降低,谷胱甘肽和SOD含量均明显升高(P<0.05)。模型组、干预组膝关节滑膜组织TACE和NF-κB蛋白表达水平较对照组均明显上调,干预组TACE和NF-κB蛋白表达明显低于模型组(P<0.05)。结论采用TNF-α抑制剂可有效减轻风湿性膝关节滑膜炎大鼠氧化应激和炎症状态,缓解膝关节损伤程度,其作用机制可能与TNF-α抑制剂可通过阻滞TACE/NF-κB信号通路而减轻大鼠风湿性膝关节滑膜炎密切相关。

Objective To analyze the effects of tumor necrosis factor-alpha(TNF-α)inhibitors on oxidative stress inhibition and downstream signaling pathways in rats with rheumatic knee synovitis.Methods Thirty-six SD rats were randomly divided into control group(normal rats),model group(rheumatic knee synovitis+saline),and intervention group(rheum atic knee synovitis+TNF-α inhibitor).Serum TNF-α,interleukin-1β(IL-1β),interleukin-6(IL-6),and matrix metalloproteinase-9(MMP-9)were assessed by enzyme-linked sorption assay.Rate scattering turbidimetry was used to detect serum C-reactive protein(CRP).Nitrite reduction method was used to detect serum superoxide dism utase(SOD)and thiobarbituric acid colorimetry simultaneously.The expression of TNF-α converting enzyme(TACE)and nuclear factor-kappa B(NF-κB)in synovial tissue of knee joint were detected by Western blot.Results Pathological examination showed that the joint tissue of model group and intervention group had obvious pathological changes.Compared with the control group,the levels of serum TNF-α,IL-1β,IL-6,MMP-9,CRP and malondialdehyde in the model group and the intervention group were significantly in creased,while the levels of glutathione and SOD were significantly reduced(P<0.05).Compared with the model group,the levels of serum TNF-α,IL-1β,IL-6,MMP-9,CRP and malondialdehyde in the intervention group were significantly reduced,and the contents of glutathione and SOD were significantly increased(P<0.05).The expression levels of TACE and NF-κB in knee synovium of model group and intervention group were significantly higher than those of control group,while the expression levels of TACE and NF-κB in intervention group were significantly lower than those in model group(P<0.05).Conclusion TNF-α inhibitors effectively alleviates oxidative stress and inflammation in rats with rheumatic knee synovitis,also the degree of knee joint injury.Its m echanism may be closely related to the blocking of TACE/NF-κB signaling pathway.