青心酮对ApoE缺陷小鼠及培养的平滑肌细胞、内皮细胞内脂素表达和分泌的影响

Effects of Dihydroxyacetophenone on the Expression of Visfatin in ApoE(-/-) Mice and the Secretion of Visfatin in Vascular Smooth Muscle Cell,Endothelial Cell

目的观察活血化瘀中药青心酮对ApoE(-/-)小鼠主动脉粥样硬化病变,血脂及斑块中主要细胞内脂素(visfatin)表达的影响。方法取普通小鼠胸主动脉,进行平滑肌细胞、内皮细胞的原代培养。实验分5组:空白对照组,IL-1β组,青心酮高剂量组,青心酮低剂量组,普伐他汀对照组。收集各组细胞上清液,应用ELISA方法检测细胞外内脂素。取8只8周龄普通小鼠做正常对照组;取24只8周龄雄性ApoE(-/-)小鼠,随机分成三组:模型组(n=8),每天生理盐水灌胃;青心酮治疗组(n=8)im给药,10 mg.kg-1.d-1;普伐他汀治疗组(n=8)ig给药,10 mg.kg-1.d-1。所有实验小鼠均饲以"西方类型膳食"饲料至12周。取血检测内脂素、血脂等,剪取主动脉根部切片、染色,电镜下观察主动脉粥样硬化斑块中平滑肌细胞、内皮细胞形态学变化,免疫组化染色法观察斑块中内脂素的分布情况。结果青心酮高剂量组血管平滑肌细胞、内皮细胞上清液中内脂素明显低于IL-1β组(P<0.01),但高于正常组;普伐他汀组亦有相似改变,但两者之间有差异(P<0.01)。在整体实验中青心酮治疗组内脂素减少、血脂降低,普伐他汀治疗组仅血脂降低,AS病灶形成皆减少,在斑块中内皮细胞、平滑肌细胞损失减轻。结论活血化瘀中药青心酮能减缓ApoE(-/-)小鼠主动脉粥样斑块的形成,可能通过抑制内脂素生成,并降低血脂发挥其作用。

OBJECTIVE To observe the effects of 3,4-dihydroxyacetophenone（DHAP） on blood lipids and plaque in ApoE（-/-） aorta atherosclerosis and the effects of DHAP on visfatin expression of vascular smooth muscle cells and endothelial cells.METHODS Normal thoracic aortas were obtained from eight week-old mice,and the VSMC and EC were cultured.The cells were divided into five groups： A.control group;B.IL-1β group;C.high dose of DHAP group;D.low dose of DHAP group;E.Pravastatin group.The changes of visfatin were analyzed by ELISA.There were 8 eight-week-old normal mice in the normal control group.24 eight-week-old male ApoE（-/-） mice were randomly divided into three groups： model group（n=8）,treated with normal saline;DHAP treatment group（n=8） treated with DHAP 10 mg·kg-1·d-1 intramuscularly;pravastatin treatment group（n=8） by gastric tube perfusion with pravastatin 10 mg·kg-1·d-1.All mice were fed with Western diet（21% fat,0.15% cholesterol） for 12 weeks.Then all mice were sacrificed by the end of the experiment.Their blood was collected for determining the concentration of blood lipids and visfatin.The morphology and composition of atherosclerotic plaques in aortic roots were examined in tissue sections.Four sections were stained with HE or immunohistochemical staining,respectively.After computer image scanning,the ratio of plaque area to luminal area were determined.RESULTS In the group treated with high dose of DHAP and pravastatin,the supernatant visfatin of vascular smooth muscle cells endothelial cells was significantly lower than that of IL-1β group（P0.01）,but higher than that of the control group.Similar changes were found in pravastatin group.There were differences between DHAP and pravastatin group（P0.01）.In vitro,the plasma visfatin and lipids decreased in DHAP treatment group,only plasma lipids decreased in pravastatin treatment group.AS lesion formation was decreased in these two groups.The injury of VSMC and EC was relieved at plaques.CONCLUSION The anti-inflammtory traditional drug——DHAP could slow down the atherosclerosis plaque formation in ApoE（-/-） mice,possibly through inhibiting the expression of visfatin and reducing blood lipids.