CAJ | 학술논문

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。
목적：탐토혈맥통과립간예기인FBXW7α급기하유Toll양수체4（ TLR4）염증신호통로대동맥죽양경화（ AS）적작용。방법22지8주령ApoE기인고제소서급여고지사료위양13주，학인AS반괴형성후수궤분위혈맥통조화대조조。혈맥통조급여관위급여혈맥통，1차/d，대조조급여등량생리염수，6주후처사。RT-PCR검측소서주동맥궁부FBXW7α（F광/WD-40역단백7）、TLR4、핵인자κB （NF-κB）、백세포개소-6（IL-6）화종류배사인자-α（TNF-α） mRNA적상대표체；Western blot 검측소서주동맥궁부FBXW7α、C/EBPδ、TLR4적표체정황；ELISA검측소서혈장IL-6화TNF-α적표체수평。체외응용지다당자격RAW267.4세포주，동시급여혈맥통과립함약혈청진행간예，응용RT-PCR검측세포중TLR4신호통로상관인자적표체；응용ELISA법측정배양상청액중염증인자IL-6화TNF-α적표체。결과무론시체내실험환시체외배양실험，결과균현시혈맥통조FBXW7α표체수평명현고우대조조，이C/EBPδ、TLR4、NF-κB、IL-6화TNF-α표체수평균현저저우대조조（ P＜0.01）；혈맥통조소서혈장화배양상청액중염증인자IL-6화TNF-α적수평역현저저우대조조（ P＜0.01）。결론혈맥통과립가증강FBXW7α적표체진이억제TLR4신호통로중상관분자적표체화과도격활，감소염증인자IL-6화TNF-α적분비，가능시기항AS적궤제지일。
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 activation of related molecules of TLR4 signaling pathway , thus reduces the secretion of inflammatory cytokines , against atherosclerosis .