血脉通颗粒对基因FBXW7α及其下游Toll样受体炎症信号通路的影响  

Effects of Xuemaitong granules on FBXW7α gene and TLR4 inflammatory signaling pathway

作　　者：张琪[1] 钱高潮[2] 丁志祥[2] 

机构地区：[1]江苏省常州市中医医院心血管科,213003 [2]江苏省常州市中医医院检验科,213003

出　　处：《广东医学》2014年第10期1491-1495,共5页Guangdong Medical Journal

基　　金：江苏省自然科学基金资助项目(编号:BK2010195)

摘　　要：目的探讨血脉通颗粒干预基因FBXW7α及其下游Toll样受体4(TLR4)炎症信号通路对动脉粥样硬化(AS)的作用。方法 22只8周龄ApoE基因敲除小鼠给予高脂饲料喂养13周,确认AS斑块形成后随机分为血脉通组和对照组。血脉通组给予灌胃给予血脉通,1次/d,对照组给予等量生理盐水,6周后处死。RT-PCR检测小鼠主动脉弓部FBXW7α(F框/WD-40域蛋白7)、TLR4、核因子κB(NF-κB)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)mRNA的相对表达;Western blot检测小鼠主动脉弓部FBXW7α、C/EBPδ、TLR4的表达情况;ELISA检测小鼠血浆IL-6和TNF-α的表达水平。体外应用脂多糖刺激RAW267.4细胞株,同时给予血脉通颗粒含药血清进行干预,应用RT-PCR检测细胞中TLR4信号通路相关因子的表达;应用ELISA法测定培养上清液中炎症因子IL-6和TNF-α的表达。结果无论是体内实验还是体外培养实验,结果均显示血脉通组FBXW7α表达水平明显高于对照组,而C/EBPδ、TLR4、NF-κB、IL-6和TNF-α表达水平均显著低于对照组(P<0.01);血脉通组小鼠血浆和培养上清液中炎症因子IL-6和TNF-α的水平亦显著低于对照组(P<0.01)。结论血脉通颗粒可增强FBXW7α的表达进而抑制TLR4信号通路中相关分子的表达和过度激活,减少炎症因子IL-6和TNF-α的分泌,可能是其抗AS的机制之一。Objective To investigate the effects of Xuemaitong granules on amelioration of atherosclerosis through TLR4 signaling pathway.Methods Male ApoE-deficient mice （n=22, aged 8 weeks, 18 to 20 g） were included, with high fat diet for 13 weeks.After the atherosclerotic plaques were confirmed , the mice were randomly divided into Xuemai-tong granules group and control group;which were sacrificed six weeks later for collection of venous blood and aorta tis -sues.The mRNA expression of FBXW7α, Toll-like receptor 4 （TLR4）, nuclear factor κB （NF-κB）, interleukin-6 （ IL-6） and tumor necrosis factor -α（ TNF-α） in aorta were measured by real -time quantitative RT-PCR.The pro-tein expression of FBXW7α, C/EBPδand TLR4 in aorta was measured by western -blot.The plasma concentrations of IL-6 and TNF-αwere measured by ELISA.RAW264.7 cell line was stimulated by lipopolysaccharide （LPS） and trea-ted with the serum of containing Xuemaitong at the same time .The mRNA expression of FBXW7α, TLR4, NF-κB, IL-6 and TNF-αin RAW264.7 was measured by real-time quantitative RT-PCR.The supernatant concentrations of IL -6 and TNF-αwere measured by ELISA .Results The mRNA expression of FBXW7αin the aorta in Xuemaitong gran-ules group was significantly higher than that in control group; however, the expression of TLR4, NF-κB, IL-6 and TNF-αin the aorta in Xuemaitong granules group was significantly lower than that in control group （P〈0.01）.The pro-tein expression of FBXW 7αin the aorta in Xuemaitong granules group was significantly higher than that in control group ;however , that of C/EBPδand TLR4 in the aorta in Xuemaitong granules group was significantly lower than that in control group （P〈0.01）.The plasma concentrations of IL -6 and TNF-αin Xuemaitong granules group was significantly lower than those in control group （P〈0.01）.The same results were obtained with RAW264.7 cell line.Conclusion Xue-maitong granules can promote the expression of FBXW 7α, and inhibit the excessive activati

关 键 词：血脉通颗粒 FBXW7α TOLL样受体4 炎症因子 动脉粥样硬化 

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