灵芝多糖对ApoE^(-/-)动脉粥样硬化小鼠TLR4/NF-κB信号通路的影响

Effect of Ganoderma Lucidum Polysaccharide on TLR4/NF-kappa B Signaling Pathway in ApoE^(-/-) Atherosclerotic Mice

目的:探讨GLPs对AS疾病TLR4信号通路调控机制的影响。方法:50只ApoE^(-/-)小鼠按随机数字表法分为模型组、GLPs低、中、高剂量组及辛伐他汀组,10只C57BL/6J小鼠作为正常组。高脂喂养12周建立AS模型,后4周各治疗组以高脂饮食结合药物方式干预,分别采用RT-PCR和western blot方法检测各组小鼠主动脉TLR4、NF-κB、My D88、TRAF-6、TRAM及TRIF等mRNA和蛋白的相对表达。结果:与正常组比较,模型组TLR4、NF-κB、My D88及TRAF-6 mRNA及蛋白表达均显著升高,GLPs干预后各指标均呈下降趋势;与模型组比较,高剂量组TLR4 mRNA和蛋白比较差异有统计学意义,低、高剂量组NF-κB mRNA及中、高剂量组NF-κB蛋白差异有统计学意义,中、高剂量组My D88 mRNA和蛋白差异有统计学意义,同时低、中、高剂量组TRAF-6mRNA及蛋白均具极显著性差异。与正常组比较,模型组TRAM、TRIF mRNA及蛋白表达均明显升高,GLPs干预后有所降低,但差异无统计学意义。结论:推测GLPs对于AS的影响可能经由TLR4通路中抑制其下游My D88依赖性信号通路起作用。

Objective: To study the effect of GLPs towards TLR4 signal transduction pathways regulating mechanism in order to explore the molecular mechanism of GLPs preventing AS. Methods: 50 ApoE-/- mice were randomly divided into model group,simvastatin group and GLPs low-dose,mid-dose,high-dose groups( n = 10);another 10 C57 BL/6 J mice served as normal group. High fat diet way was giving to ApoE-/- mice for 12 weeks to establish AS model. Then the ApoE-/- mice of drug therapy groups had been fed with high fat diet combined with drugs for 4 weeks. On the basis of model successfully established,the Real-time fluorescent quantitative PCR and western blot methods were used to respectively detecting mRNA and protein of TLR4,NF-κB,MyD88,TRAF-6,TRAM and TRIF in mouse aortic. Results: The mRNA and protein relative expressing were activated of TLR4 and primary element of downstream MyD88 dependent signal transduction pathway in model group,such as TLR4,NF-κB,MyD88 and TRAF-6,which had significant differences compared with normal group. The GLPs could restrain mRNA and protein expressing of TLR4,NF-κB,MyD88 and TRAF-6 after being interfered in on different levels. The TLR4 mRNA and protein for GLPs-High had significant differences compared with model group. The GLPs-Low,GLPs-High NF-κB mRNA and GLPs-Mid,GLPs-High NF-κB protein had significant differences compared with model group. The MyD88 mRNA and protein for GLPs-Mid and GLPs-High also had significant differences compared with model group. And then,the TRAF-6 mRNA and protein of all GLPs groups had highly significant differences compared with model group. The mRNA and protein expressing were activated of TLR4 and primary element of downstream MyD88 independent signal transduction pathway in model group,such as TRAM and TRIF,which had significant differences compared with normal group. The GLPs had no obvious restraining fuction of mRNA and protein expressing of TRAM and TRIF after being interfered in. Conclusion: It was speculated that GLPs might restrain mRNA and protein expressing of TLR4 and primary element of downstream MyD88 dependent signal transduction pathway.