摘要
目的:探究独活寄生汤对佐剂型关节炎大鼠血清TLR4/NF-κB信号通路的调控机制。方法:SPF级Wistar大鼠共90只,按照随机数字法分为空白对照组、模型对照组、阳性对照组、独活寄生汤治疗高、中、低剂量组共6组,每组大鼠各15只。实验组大鼠均采取足跖内部位皮下注射0.1mL弗氏完全佐剂建立佐剂型关节炎大鼠。造模成功后5d,分别按照剂量4 g·kg^-1、16 g·kg^-1、8 g·kg^-1对佐剂型关节炎低、高、中剂量三组大鼠进行给药灌胃;阳性对照组大鼠给予甲氨蝶呤2.7mg·kg-1灌胃治疗;空白对照组及模型对照组大鼠给予等容积5mL蒸馏水进行灌胃,各组每天均给药1次,连续给药3周。末次给药24h后,酶联免疫吸附法(ELISA)测定各组大鼠外周血TNF-α(Tumor necrosis factor-α,肿瘤坏死因子α)、IL-4(interleukin-4,白介素4)、IL-6(interleukin-6,白介素6)表达水平;蛋白免疫印迹法(Western blotting法)检测不同组大鼠关节组织中TLR4,p-NF-κB蛋白的表达变化。结果:造模后,各造模组大鼠AI积分明显增高,且外周血清中TNF-α、IL-4、IL-6炎性因子的含量明显增长(P<0.05);关节组织中TLR4、p-NF-κB蛋白表达水平明显升高(P<0.05)。与模型对照组相比,独活寄生汤治疗组及阳性对照组的大鼠在治疗后AI积分明显降低,而且外周血清中TNF-α、IL-4、IL-6含量明显降低(P<0.05);关节组织中TLR4、p-NF-κB蛋白表达水平显著降低(P<0.05),且独活寄生汤治疗组大鼠具有剂量依赖性(P<0.01)。结论:独活寄生汤可有效调控佐剂型关节炎大鼠血清中TNF-α、IL-4、IL-6炎性因子含量,调节TLR4/NF-κB信号蛋白标表达,对佐剂型关节炎大鼠关节组织起到有效保护作用。
Objective:To explore the regulatory mechanism of Duhuo Jisheng decoction on serum TLR4/NF-κB signaling pathway in adjuvant arthritis rats.Method:A total of 90 SPF Wistar rats were divided into 6 groups according to random number method:blank control group,model control group,positive control group,and Duhuo Jixiang Decoction treatment high,medium and low dose groups,15 rats in each group only.Rats in the experimental group were injected with 0.1 mL of Freund’s complete adjuvant subcutaneously in the foot plantar to establish adjuvant arthritis rats.Five days after the successful modeling,the three groups of rats with adjuvant arthritis low,high,and medium doses were administered intragastrically according to the doses of 4 g·kg^-1,16 g·kg^-1,and 8 g·kg^-1 respectively;The rats in the positive control group were given methotrexate 2.7 mg·kg^-1 by gavage;the rats in the blank control group and the model control group were given an equal volume of 5 mL distilled water for gavage,and each group was given once a day for continuous administration 3 weeks.24 hours after the last administration,the TNF-α(Tumor necrosis factor-α,tumor necrosis factorα)and IL-4(interleukin-4,interleukin-4)in the peripheral blood of rats in each group were measured by enzyme-linked immunosorbent assay(ELISA),IL-6(interleukin-6)expression level;Western blotting method(Western blotting)was used to detect the expression changes of TLR4 and p-NF-κB protein in the joint tissues of different groups of rats.Results:After modeling,the AI scores of rats in each model increased significantly,and the levels of inflammatory factors such as TNF-α,IL-4,and IL-6 in peripheral serum increased significantly(P<0.05);TLR4,IL-6 in the joint tissues The expression level of p-NF-κB protein was significantly increased(P<0.05).Compared with the model control group,the AI scores of rats in the Duhuo Jisheng decoction treatment group and the positive control group were significantly reduced after treatment,and the levels of TNF-α,IL-4,and IL-6 in perip
作者
范卫闯
吕立桃
FAN Weichuang;LV Litao(Kaifeng People’s Hospital,Henan Kaifeng 475002,China)
出处
《中医药临床杂志》
2020年第8期1478-1482,共5页
Clinical Journal of Traditional Chinese Medicine
基金
山东省中医经典名方协同创新中心开放课题(No.2018KFY26)。
