基于TLR4/NF-κB炎性轴的白芥子经皮给药对类风湿性关节炎大鼠调控机制研究

Regulatory Mechanism of Semen sinapis Transdermal Administration on Rheumatoid Arthritis in Rats Based on TLR4/NF-κB Inflammatory Axis

目的:探讨白芥子经皮给药通过Toll样受体4/核因子-κB(TLR4/NF-κB)炎性轴对类风湿性关节炎(RA)大鼠的治疗作用。方法:50只大鼠随机分为对照组(空白对照组)、模型组、阳性药组、白芥子组及白芥子+TLR4激活剂组,每组10只。除对照组外,其余组制备佐剂性RA模型。阳性药组右后足涂抹0.27 g/kg扶他林,白芥子组涂抹4.50 g/kg白芥子涂剂,白芥子+TLR4激活剂组涂抹4.50 g/kg白芥子涂剂同时踝关节注射0.25 mg/kg TLR4激活剂LPS,对照组和模型组涂抹等量的面粉糊作对照,制备模型前3 d开始给药,连续18 d。测量不同时间左右后足肿胀度,末次给药后检测热痛和机械痛阈值、血清白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α),Western blot法检测踝关节滑膜组织中TLR4、NF-κB蛋白表达情况。结果:与模型组比较,阳性药组、白芥子组及白芥子+TLR4激活剂组建模不同时间右后足肿胀度均降低,其中阳性药组和白芥子组均低于白芥子+TLR4激活剂组,白芥子组建模9 d右后足肿胀度低于阳性药组(P<0.05)。与模型组比较,阳性药组、白芥子组及白芥子+TLR4激活剂组右后足热痛及机械痛阈值均升高,其中阳性药组高于白芥子组,白芥子组高于白芥子+TLR4激活剂组(P<0.05)。与模型组比较,阳性药组、白芥子组及白芥子+TLR4激活剂组血清IL-1β、TNF-α水平及踝关节滑膜组织中TLR4、NF-κB蛋白相对表达量均降低,其中阳性药组和白芥子组低于白芥子+TLR4激活剂组(P<0.05),阳性药组与白芥子组比较,差异无统计学意义(P>0.05)。结论:白芥子经皮给药通过抑制TLR4/NF-κB炎性轴发挥作用,可有效治疗RA模型大鼠。

Objective:To investigate the therapeutic effect of Semen sinapis through the Toll-like receptor 4/nuclear factor-κB(TLR4/NF-κB)inflammatory axis on rheumatoid arthritis(RA)rats by transdermal administration.Methods:50 rats were randomly divided into the control group,the model group,the positive drug group,the Semen sinapis group and the Semen sinapis plus TLR4 activator group,with 10 rats in each group.Except for the control group,the remaining groups were prepared adjuvant RA models.In the positive drug group,voltarin(0.27 g/kg)was applied to the right hind foot,the Semen sinapis group was applied with Semen sinapis lotion(4.50 g/kg),and the Semen sinapis plus TLR4 activator group was treated with Semen sinapis lotion(4.50 g/kg)and LPS(0.25 mg/kg).The control group and the model group were smeared with the flour paste,and the administration was started 3 days before the preparation of the model for 18 consecutive days.The swelling of the left and right hind feet at different times was measured.After the last administration,thermal pain and mechanical pain thresholds,serum interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)were detected.The protein expressions of TLR4 and NF-κB in the ankle joint synovial tissues were detected by Western blot method.Results:Compared with that in the model group,the swelling of the left and right hind feet at different modeling time was reduced in the positive drug group,the Semen sinapis group and the Semen sinapis plus TLR4 activator group;the swelling of the positive drug group and the Semen sinapis group were lower than that of the Semen sinapis plus TLR4 activator group;the right hind foot swelling on the 9th day of modeling was lower in the Semen sinapis group than that in the positive drug group(P<0.05).Compared with those in the model group,the thresholds of thermal pain and mechanical pain of the right hind foot were increased in the positive drug group,the Semen sinapis group and the Semen sinapis plus TLR4 activator group;which were higher in the positive drug group than those in the Semen sinapis group,and they were higher in the Semen sinapis group than those in the Semen sinapis group plus TLR4 activator group(P<0.05).Compared with those in the model group,the serum levels of IL-1βand TNF-α,as well as the relative protein expressions of TLR4 and NF-κB in the ankle joint synovial tissues were decreased in the positive drug group,the Semen sinapis group and the Semen sinapis plus TLR4 activator group;which were lower in the positive drug group and the Semen sinapis group than those in the Semen sinapis plus TLR4 activator group(P<0.05);in which there were no statistical differences in the positive drug group and the Semen sinapis group(P>0.05).Conclusion:The transdermal administration of Semen sinapis can effectively treat RA rats,possibly by inhibiting the TLR4/NF-κB inflammatory axis.