熊胆粉在麝香通心滴丸抗动脉粥样硬化中的作用机制研究  

Anti-atherosclerotic Effects of Bear Bile Powder in Shexiang Tongxin Dripping Pill:a Mechanism Study

作　　者：熊敏琪[1,2] 贾成林[1,2] 崔金刚[1,2] 宁冰冰[1,2] 朱玉陵 王文健[1,2,4] 陈瑜[1,2] 张腾[1,2] 

机构地区：[1]上海中医药大学附属岳阳中西医结合医院,上海200437 [2]上海市中医药研究院中西医结合临床研究所,上海200437 [3]内蒙古康恩贝药业有限公司,上海201318 [4]复旦大学中西医结合研究所,上海200040

出　　处：《中国中西医结合杂志》2015年第9期1083-1089,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然基金面上项目(No.81273960);国家中医药管理局中西医结合临床重点学科建设项目(国中医药发[2009]30号);上海市中西医结合学会课题(No.ZXYK-1246);上海市高校特聘教授(东方学者)人才计划(沪教委人[2010]84号);沪教委人([2011]88号);上海市浦江计划(No.11PJ1409000;No.13PJ1407800);上海市中医药事业三年行动计划项目(No.ZYSNXD-CC-ZDXK-09);上海市085一流学科建设科技创新支持计划(No.085ZY1212;No.085ZY1221)

摘　　要：目的研究熊胆粉在麝香通心滴丸抗动脉粥样硬化的作用机制,为麝香通心滴丸"凉开"法治疗动脉粥样硬化性疾病提供科学依据。方法采用ApoE^(-/-)基因敲除小鼠结合高脂饮食复制动脉粥样硬化动物模型,30只小鼠按体重用随机数字表法分为模型组(9只),麝香通心滴丸组(复方组,7只),麝香通心滴丸去熊胆粉组(拆方组,7只),熊胆粉组(7只);另取9只雄性同龄C57BL/6J小鼠设为正常对照组。复方组、拆方组和熊胆粉组分别给予对应药物(30、30、0.33 mg/kg)灌胃,模型和正常对照组予生理盐水灌胃,连续8周。E LISA法检测血清IL-2、IL-6、肿瘤坏死因子(tumor necrosis factor-α,TNF-α)、γ-干扰素(interferon-γ,IFN-γ)及氧化性低密度脂蛋白(oxygenized low density lipoprotein,oxLDL)含量;采用生化法测定血清谷胱甘肽(glutathione,GSH)含量、血清超氧化物歧化酶(superoxide dismutase,SOD)活性和丙二醛(malondialdehyde,MDA)含量。二氢乙啶(dihydroethidum,DHE)荧光探针探测活性氧(reactive oxygen species,ROS)含量;Real-time PCR检测主动脉miR-20、miR-21、miR-126及miR-155表达。结果模型组小鼠主动脉荧光强度明显增强,复方组、拆方组和熊胆粉组荧光强度明显减弱,以复方组减弱的最为明显。与正常组比较,模型组小鼠血清IL-2、IL-6、TNF-α、IFN-γ、oxLDL和MDA均升高(P<0.01),GSH和SOD下降(P<0.01),主动脉miR-126、miR-21及miR-1 55表达上升(P<0.01),miR-20表达下降(P<0.01)。与模型组比较,复方组、拆方组和熊胆粉组血清IL-2、IL-6、TNF-α、IFN-γ、oxLDL及MDA下降(P<0.01),GSH和SOD上升(P<0.01),主动脉miR-126、miR-21、miR-155表达下调(P<0.01),复方组和熊胆粉组miR-20表达上调(P<O.01)。与复方组比较,拆方组和熊胆粉组小鼠血清中IL-2、IL-6、TNF-α及IFN-γ水平升高(P<0.01);拆方组和熊胆粉组GSH和SOD下降(P<0.01),MDA和oxLDL上升(P<0.01);拆方组与熊胆粉组miR-20和miR-155表达下调(P<0.01)。结论麝香通Objective To study the anti-atherosclerotic mechanism of bear bile powder（BBP）in Shexiang Tongxin Dripping Pill（STDP）,and to provide scientific evidence for treating atherosclerosis（AS） by its therapeutic characteristics of cool resuscitation.Methods AS model was duplicated using ApoE^（-/-） gene knocked mice fed with high-fat diet.Thirty ApoE^（-/-） deficient male mice were divided into four groups according to body weight using random digit table,i.e.,the model group（A,n = 9）,the STDP group（B,n =7）,the STDP without BBP group（C,n =7）,and the BBP group（D,n =9）.Besides,another 9 C57BL/6J male mice of the same age were recruited as a normal control group（E）.All mice in Group B,C,and D were respectively administered with corresponding drugs（30,30,and 0.33 mg/kg）by gastrogavage.Equal volume of normal saline was administered to mice in Group A and E.All medication lasted for 8 successive weeks.Serum levels of inflammatory cytokines such as interleukin 2（IL-2）,interleukin 6（IL-6）,tumor necrosis factor α（TNF-α）,interferon γ（IFNγ）,and oxidized low-density lipoprotein（ox-LDL） were measured by ELISA.Serum levels of malondialdehyde（MDA）,activities of glutathione（GSH） and superoxide dismutase（SOD） were determined using biochemical assay.Contents of reactive oxygen species（ROS） in the aortic root was detected by dihydroethidum（DHE） fluorescent probe.Expression levels of microRNAs（such as miR-20,miR-21,miR-126,and miR-155） were detected by real-time PCR.Results The fluorescence intensity of the aorta was obviously enhanced in Group A.But it was obviously attenuated in Group B,C,and D,and the attenuation was the most in Group B.Compared with Group E,serum levels of IL-2,IL-6,TNF-α,IFN-γ,oxLDL,and MDA all increased（P 0.01）,GSH contents and SOD activities decreased（P 0.01）,expression levels of miR-126,miR-21,and miR-155 in aorta increased（P 0.01）,and the expression level of miR-20 decreased in Group A（P0.01）.Compared with G

关 键 词：麝香通心滴丸 熊胆粉 动脉粥样硬化 氧化应激 炎症 微小RNA 

分 类 号：R285.5[医药卫生—中药学]