青藤碱对胶原诱导性关节炎小Toll样受体4/髓样分化因子88通路的影响

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目的 探讨青藤碱(sinomenine,SN)对胶原诱导性关节炎(collagen-induced arthritis,CIA)小Toll样受体4(toll like receptor 4,TLR4)/髓样分化因子88(myeloid differentiation factor 88,MyD88)信号通路诱导炎症的影响.方法 6周龄DBA/1小30只,随机分为正常对照组、CIA组及SN组,每组10只.其中CIA组及SN组小鼠尾根部初次免疫及加强免疫Ⅱ型胶原乳剂诱导CIA模型.小鼠初次免疫d28开始灌胃给药,SN组予SN治疗,正常对照组和CIA组予生理盐水作为对照.连续治疗20 d后处死小鼠,观察药物对小鼠关节肿胀的影响,收集膝关节滑膜及血清.Western blot法检测滑膜TLR4及MyD88蛋白表达,酶联免疫吸附试验(enzyme-linked immunosorbent assays,ELISA)检测血清炎性因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的水平.结果 经治疗后,SN组小鼠关节肿胀情况较CIA组改善明显,差异有统计学意义[d41(5.50±1.38)比(9.67±2.34),P<0.05;d48(1.67±0.52)比(3.33±1.21),均P<0.05];TLR4、MyD88蛋白表达及血清炎性因子TNF-α均较CIA组下降,差异有统计学意义[TLR4/β-actin:(0.33±0.05)]比(0.62±0.05)];MyD88/β-actin:(0.10±0.02)比(0.33±0.03);TNF-α:(169.16±10.68)比(347.97±13.45),均P<0.05].结论 SN可改善CIA小鼠的关节炎症,该作用可能与其抑制TLR4/MyD88信号通路有关.“,”Objective To investigate the effects of Sinomenine(SN)on TLR4 / MyD88 signal pathway in mice with collagen-induced arthritis(CIA). Methods 30 DBA / 1 mice of 6-week-old were randomly divid-ed into normal control group,CIA group,and SN group. There were 10 mice in each group. CIA was induced in the mice from the CIA group and the SN group by primary and secondary immunization with Ⅱ collagen emul-sion on the base of the mouse tails. The day of the primary immunization was referred to as day 0(d0). The mice in the SN group were given SN intragastrically once a day from d28,while mice in the control group and CIA group were given saline. All mice were treated for 20 days continuously. Arthritis index(AI)scores were meas-ured and recorded based on joint swelling. After treated for 20 days,knee synovium of the mice were collected to detect the expression of toll like receptor 4(TLR4)and MyD88 by Western blot. The level of tomor necrosis fac-to-α(TNF-α)in the serum was determined by enzyme-linked immunosorbent assays(ELISA). Results After treatment,the joint swelling of SN group was significantly improved compared with that of CIA group;the differ-ence was statistically significant[d41:(5. 50 ± 1. 38)vs(9. 67 ± 2. 34);d48:(1. 67 ± 0. 52)vs(3. 33 ± 1. 21), P < 0. 05]. TLR4 and MyD88 in synovial tissue and inflammatory factor TNF-α of the serum were significantly decreased in the mice treated with SN,compared with CIA control mice;the difference was statistically signifi-cant[TLR4 / β-actin:(0. 33 ± 0. 05)vs(0. 62 ± 0. 05)];MyD88 / β-actin:(0. 10 ± 0. 02)vs(0. 33 ± 0. 03);TNF-α:(169. 16 ± 10. 68)vs(347. 97 ± 13. 45),all P < 0. 05]. Conclusion Sinomenine can improve mouse arthritis induced by collagen through the mechanism related to the inhibition of TLR4 / MyD88 signaling path-way.
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