青心酮防治动脉粥样硬化与TLR4信号转导途径的关系  

Relationship Between Therapeutic and Preventive Effect of Dihydroxyacetophenone in Treating Atherosclerosis and the Toll-like-receptor 4 Pathway

作　　者：张代娟[1] 刘同美[1] 刘江月[1] 崔晓栋[1] 郭军堂[1] 王建英[1] 叶笃筠[2] 

机构地区：[1]潍坊医学院病理生理教研室,山东潍坊261053 [2]华中科技大学同济医学院病理生理教研室,武汉430030

出　　处：《中药新药与临床药理》2009年第5期404-407,共4页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：山东省中医药科技发展计划项目(2005-238)

摘　　要：目的观察青心酮对载脂蛋白E基因敲除小鼠主动脉粥样硬化病变的影响及其与平滑肌细胞、内皮细胞、巨噬细胞Toll样受体4T(oll-like receptor4,TLR4)信号转导途径的关系。方法取小鼠胸主动脉,进行平滑肌细胞、内皮细胞的原代培养,巨噬细胞RAW 264.7细胞株传代培养。实验分4组,即空白对照组(正常细胞培养,加入等数量的培养基,不加入任何药物),脂多糖组L(PS组)(加入等数量的培养基3h,再加入LPS10ng.mL-1刺激8h),DHAP组(先加入溶有DHAP 1×10-7mol.L-1的培养基培养3h,再加入LPS 10ng.mL-1刺激8h),辛伐他汀对照组(先加入溶有辛伐他汀5×10-7m ol.L-1的培养基培养3h,再加入LPS 10ng.mL-1刺激8h)。收集各组细胞蛋白,应用Western-blot检测TLR4蛋白含量。收集上清液,用ELISA法检测TNF-α。取8周龄雄性ApoE(-/-)小鼠20只,随机分成两组,每组10只,模型对照组建立小鼠AS模型,青心酮治疗组经胃管灌注青心酮10mg.kg-1.d-1。所有实验小鼠均饲以"西方类型膳食"饲料喂养12周。剪取主动脉根部行石蜡切片及HE染色,观察主动脉粥样硬化的病变情况。结果青心酮组血管平滑肌细胞、内皮细胞、巨噬细胞TLR4蛋白含量均明显低于未处理组(P<0.01),但高于空白对照组;与辛伐他汀组比较无显著性差异;青心酮组TNF-α含量明显低于未处理组(P<0.01),但高于空白对照组;与辛伐他汀组比较,具有显著性差异(P<0.01);青心酮组AS病灶形成减少。结论青心酮能够在一定程度上减少AS病灶的形成,可能是通过TLR4信号转导途径发挥作用的。Objective To observe the therapeutic effect of dihydroxyacetophenone （DHAP） on aortic atherosclerosis （AS） in mice with apolipoprotein gene E knockoff, and to explore the relationship between DHAP in treating arotic atherosclerosis and the Toll - like receptor 4 （TLR4） pathway in vascular smooth muscle cells （VSMC）, endothelial cells （EC） and macrophages （MC）. Methods Normal thoracic aortas were obtained from 4 adult mice, and theirVSMC and EC were cultured. The MC （RAW264. 7 cell lines） was also cultured. Cells were divided into four groups： blank control group （routine culture media without any medicine）, LPS group（stimulating with LPS 10 ng ·mL^-1 for 8 h after routine culturing for 3 h）, DHAP group（stimulating with LPS 10 ng ·mL^-1 for 8 h after the culturing with DHAP 10^-7 mol·L^-1 for 3 h）, and Simvastatin group （stimulating with LPS 10 ng · mL^-1 for 8 h after the culturing with Simvastatin 5×10^-7 mol·L^-1 for 3 h）. Changes of TLR4 protein were analyzed by Western blot and the changes ofFNF- α detected by ELISA. In addition, 20 ApoE（ - / - ） male mice at same age were fed with a Western diet （21% fat and 0. 15 % cholesterol） for 12 weeks to induce AS. These AS mice were divided into two groups at random： model group and DHAP group（gastric garage of DHAP 10 mg·kg^-1· d^-1 for consecutive 4 weeks）. The pathological changes of aortic atherosclerosis were analyzed by HE staining. Results TLR4 protein content in the VSMC, EC and MC of DHAP group was lower than that in LPS group（ P 〈 0.01 ）, and higher than that in the blank control group, but did not differ from that in Simvastatin group ; the secretion of TNF - α in DHAP group was lower than that in LPS group（ P 〈 0.01）, higher than that in the blank control group, and differed from that in Simvastatin group（ P 〈 0. 01 ）. The lesions of aortic atheroscle- rosis were inhibited in DHAP group. Conclusion It is suggested that DHAP could be effective for the prevention and tre

关 键 词：青心酮 动脉粥样硬化 TOLL样受体4 载脂蛋白E基因敲除小鼠 

分 类 号：R285.5[医药卫生—中药学]