葡萄籽原花青素对稳定apoE基因敲除小鼠易损斑块的影响及机制  

Effects of grape seed proanthocyanidins on stability of atherosclerotic plaques in apoE^-/-mice

作　　者：贾敏[1] 路永刚[1] 崔秋英[1] 高伟[1] 帖彦清[1] JIA Min;LU Yonggang;CUI Qiuying;GAO Wei;TIE Yanqing(Hebei General Hospital,Shijiazhuang 050051,China)

机构地区：[1]河北省人民医院,石家庄050051

出　　处：《山东医药》2020年第20期33-35,共3页Shandong Medical Journal

基　　金：河北省卫生健康委医学科学研究课题指令性计划(20160068)。

摘　　要：目的观察葡萄籽原花青素(GSP)对apoE基因敲除(apoE^-/-)小鼠易损动脉粥样硬化(AS)斑块的影响,并探讨其机制。方法24只apoE^-/-小鼠随机分为GSP低剂量组、GSP中剂量组、GSP高剂量组和模型组,每组6只。GSP低、中、高剂量组分别予以GSP溶液45、90、180 mg/(kg·d)灌胃,模型组给予同体积生理盐水灌胃。各组均干预8周后,取小鼠降主动脉,HE染色测算斑块面积,采用免疫组化及特殊染色法测算主动脉根部斑块内容物包括巨噬细胞(棕色)和胶原(红色)阳性区域面积。采用DHE荧光染色法检测主动脉根部活性氧(ROS)水平。采用TUNEL法测算斑块内TUNEL阳性面积,反映细胞凋亡情况。采用Western blotting法检测降主动脉凋亡相关蛋白细胞色素C(CytC)、cleaved Caspase-3和Bax的表达。结果GSP高剂量组主动脉根部斑块面积、巨噬细胞面积均低于模型组,胶原面积高于模型组(P均<0.05)。GSP高剂量组主动脉根部斑块内红色荧光面积及TUNEL阳性面积均低于模型组(P均<0.01)。GSP高剂量组促凋亡蛋白Cytc、cleaved Caspase-3和Bax表达均低于模型组(P均<0.05)。结论GSP 180 mg/kg对小鼠主动脉AS斑块具有稳定易损斑块、减小斑块面积、抑制巨噬细胞聚集及细胞凋亡作用,其机制可能与抑制ROS产生、减少促凋亡蛋白表达有关。Objective To investigate the effects and mechanisms of grape seed proanthocyanidin(GSP)on vulnerable plaques of apoE^-/-mice.Methods Twenty-four apoE^-/-mice were randomly divided into four groups:the model group and low-dose,medium-dose,high-dose GSP groups,which were then gavaged with normal saline and 45,90,180 mg/kg GSP,respectively,and meanwhile,they were fed with atherogenic fat diet for 8 weeks to induce the vulnerable plaques and at the end of experiment the descending aortas were dissected.HE,immunohistochemistry and special staining were used to detect the plaque areas and component macrophages and collagen included.Dihydroethidium(DHE)and TdT-mediated dUTP nick-end-labeling(TUNEL)staining were used for valuing oxidative stress and apoptosis;apoptosis-associated protein cytochrome C(CytC),Cleaved-Caspase-3 and Bax were tested by Western blotting.Results High-dose GSP suppressed aortic roots lesional areas,macrophage positive areas,and increased collagen content(all P<0.05),and decreased TUNEL as well as DHE-positive areas(all P<0.01);the expression levels of pro-apoptotic protein Cytc,Cleared-Caspase-3 and Bax in the high-dose GSP group were lower than those in the model group(all P<0.05).Conclusion GSP(180 mg/kg)stabilized vulnerable lesions,suppressed lesional areas and reduced macrophages accumulation and cellular apoptosis by inhibiting oxidative stress and decreasing pro-apoptosis protein expression..

关 键 词：动脉粥样硬化 葡萄籽原花青素 氧化应激 凋亡 

分 类 号：R972[医药卫生—药品]