银杏酮酯对心肌细胞Toll样受体4/核转录因子κB及血管紧张素原、血管紧张素Ⅱ1型受体的抑制作用  

Effect of Ginkgo Biioba Extract 50 on TLR4/NF-κB Signaling Mediated Expression of Angiotensinogene and Angiotensin Ⅱ Type 1 Receptor in Cardiac Myocyte

作　　者：姜华[1] 曲鹏[1] 崔晓琼[2] 王虹艳[1] 王纪文[1] 李桂华[1] 

机构地区：[1]大连医科大学附属二院心内科,辽宁大连116027 [2]天津市第三中心医院心内科,天津300170

出　　处：《中华高血压杂志》2008年第11期1013-1017,共5页Chinese Journal of Hypertension

基　　金：国家自然科学基金赞助项目(No.30371558)~~

摘　　要：目的观察银杏酮酯50(GBE50)对脂多糖激活的心肌细胞 Toll 样受体4(TLR4)/核转录因子κB(NF-κB)信号及血管紧张素原(ATG)、血管紧张素Ⅱ 1型受体(AT_1R)表达的影响,探讨 GBE50防治左室重构的作用机制。方法体外培养新生大鼠心肌细胞(NRVM),分4组:1)对照组:DMEM 培养基中不加任何干预因素;2)脂多糖组:DMEM 培养基中加入脂多糖1 mg/L;3)脂多糖+GBE50组:预先加入 GBE50 80 mg/L,1 h 后加入脂多糖1 mg/L;4)脂多糖+咖啡酸苯乙酯组:预先加入咖啡酸苯乙酯20 mg/L30 min 后加入脂多糖1mg/L。干预24 h 后免疫细胞化学方法观察 NF-κB p65核易位情况;RT-PCR 检测 TLR4、ATG 及 AT_1R、心肌肌球蛋白重链β-MHC mRNA 的表达情况,考马斯亮蓝法测定心肌细胞蛋白含量。结果 LPS 刺激下 NRVM TLR4 mR-NA 明显升高(P<0.01),NF-κB p65核易位增多(P<0.01),ATG 和 AT_1R、心肌肌球蛋白重链β-MHC mRNA表达以及细胞蛋白含量明显增高(P<0.01)。GBE50与咖啡酸苯乙酯能明显抑制 NF-κB p65核易位,同时抑制LPS 诱导的 TLR4、ATG 和 AT_1Rβ-MHCmRNA 表达上调以及细胞蛋白含量的增加(P<0.05)。结论 GBE50可能通过抑制 NF-κB活化,进而减弱 TLR4/NF-KB 信号从而抑制心肌 RAS 的激活,干预 FLR4/NF-κB信号通路可能是银杏叶提取物防治心肌肥大的机制之一。Objective To investigate the effect of Chinese traditional medicine, Ginkgo biloba extract （GBES0） on TLR4/NF-κB signaling and expression of angiotensinogene and angiotensin Ⅱtype 1 receptor （AT1R） activated by lipopolysaccharide（LPS） in cardiac myocytes. Methods Cultured neonatal rat myocytes （NRVM） were randomized to receive following approaches： LPS 1 mg/L; pretreated with GBE50 80 mg/L for 1 hour before LPS incubation; pretreated with phenethyl ester（a specific inhibitor of NF-κB） 20 mg/L for 30 min before LPS incubation. NF-κB p65 was detected by immunocytochemistry. The quantification of TLR4, angio-tensinogen, AT1R mRNA and β-MHC expression were assessed by reverse transcriptase-polymerase chain reaction （RT-PCR）. The protein contents of cardiac myocytes was determined by brilliant blue method. Results Both GBE50 and CAPE inhibited NF-κB activation in cultured NRVM induced by LPS 1 mg/L（P〈0.01）, and sup- pressed the upregulation of TLR4, angiotensinogen, AT1R and β-MHC mRNA expression and increases in protein content（P〈0.01）. Conclusion GBE50 inhibited NF-κB acitivation and subsequently suppressed TLR4/NF-κB signal and renin-angiotensin system, which may be one of the mechanisms of GBES0 preventing cardiac hypertrophy.

关 键 词：TOLL样受体4 核转录因子ΚB 脂多糖 新生大鼠心肌细胞 银杏叶提取物 

分 类 号：R541[医药卫生—心血管疾病]