摘要
目的:考察当归四逆汤对类风湿性关节炎模型大鼠血清中炎性因子表达的影响,并初步探究其通过TLR/Beclin-1信号通路影响类风湿性关节炎的调控机制。方法:SPF级Wistar大鼠共60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、当归四逆汤治疗高、中、低剂量组共6组,每组大鼠各10只。实验组大鼠均采取足跖内部位皮下注射0.1 mL弗氏完全佐剂建立类风湿性关节炎模型大鼠。造模成功后5 d,分别按照剂量4、16、8 g·kg-1剂量灌胃给予模型动物当归四逆汤;阳性对照组大鼠给予甲氨蝶呤2.7 mg·kg-1灌胃治疗;空白对照组及模型对照组大鼠给予等容积5 mL蒸馏水灌胃,各组每天均给药1次,连续给药3周。末次给药24 h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-1β(interleukin-1β,白介素1β)、IL-6(interleukin-6,白介素6)表达水平;HE染色检测不同组大鼠的关节组织病理表现;蛋白免疫印迹法(Western blotting法)检测不同组大鼠关节组织中TLR,Beclin-1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-1β、IL-6炎性因子的含量明显增长(P<0.05);关节组织中TLR、Beclin-1蛋白表达水平明显升高(P<0.05)。与模型对照组相比,当归四逆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-1β、IL-6含量明显降低(P<0.05);关节组织中TLR、Beclin-1蛋白表达水平显著降低(P<0.05),且当归四逆汤治疗组大鼠具有剂量依赖性(P<0.01)。结论:当归四逆汤可有效调控类风湿性关节炎模型大鼠血清中TNF-α、IL-1β、IL-6炎性因子含量,调节TLR/Beclin-1信号蛋白标表达,对类风湿性关节炎模型大鼠关节组织起到有效保护作用。
Objective:To observe the effect of Danggui Sini Decoction on the expression of inflammatory factors in serum of rats with rheumatoid arthritis,and to investigate the regulatory mechanism on rheumatoid arthritis through TLR/Beclin-1 signaling pathway.Method:According to random number,60 SPF-grade wistar rats were divided into blank control group,model control group,positive control group,Danggui Sini Decoction high,medium and low dose group,10 rats in each group.Rats in the experimental group were treated with 0.1 mL Freund’s complete adjuvant in the internal subcutaneous injection of vola pedis to establish a rheumatoid arthritis model.Rats in Danggui Sini Decoction group were given gastric perfusion at doses of 4 g·kg-1,16 g ·kg-1 and 8 g·kg-1,respectively.Rats in positive control group were given intragastric administration of methotrexate 2.7 mg·kg-1.5 mL of distilled water was given in blank control group and model control group.Each group was given once a day for 3 weeks.After the last 24 h of administration,enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of peripheral blood tumornecrosis factor-α (TNF-α),interleukin-1β (IL-1β),interleukin-6 (IL-6) in each group.HE staining was used to detect the pathological expression of joint tissue in different groups,and Western Blot method was used to detect the protein expression of TLR Beclin-1 in rats joint of different groups.Result:After the establishment of the model,the AI score of the rats in each model was significantly increased.The content of TNF-α,IL-1 β,IL-6 inflammatory factors in the peripheral serum was significantly increased (P < 0.05).The protein expression level of TLR and Beclin-1 in the joint tissue was significantly increased (P < 0.05).Compared with model control group,the AI scores of the rats in the treatment group and the positive control group were significantly reduced,and the contents of TNF-α,IL-1 β,IL-6 in the peripheral serum were significantly reduced (P < 0.05).The expression protein levels of TLR and Beclin-1 in the joint tissue were significantly reduced (P< 0.05).The effect of the treatment group were dose-dependent (P < 0.01).Conclusion:Danggui Sini Decoction can effectively regulate the content of TNF-α,IL-1 β,IL-6 inflammatory factors in the serum of rats with rheumatoid arthritis,regulate the expression of TLR/Beclin-1 signal protein,and play an effective role in protecting the joint tissue of rats with rheumatoid arthritis.
作者
范卫闯
吕立桃
FAN Wei-chuang;LV Li-tao(Department of Bones,Kaifeng People's Hospital,Kaifeng 475002,Henan)
出处
《中药与临床》
2020年第2期27-30,共4页
Pharmacy and Clinics of Chinese Materia Medica
基金
中医经典名方协同创新中心开放课题(No.2018KFY26)。
