摘要
目的:观察阿托伐他汀(Atorv)对链脲佐菌素(STZ)诱导的糖尿病高脂喂养载脂蛋白E敲除(apolipoprotein E knockout,ApoE-/-)小鼠动脉粥样硬化的影响,探讨阿托伐他汀在糖尿病合并高脂饮食条件下对抗动脉粥样硬化的机制。方法:C57小鼠8只作为对照,34只高脂喂养的ApoE-/-小鼠随机分为3组:ApoE-/-组、STZ-ApoE-/-组和STZ-ApoE-/-+Atorv组。STZ腹腔注射建立糖尿病动物模型,测定小鼠空腹血糖、血脂水平,HE染色图像分析测定胸主动脉斑块面积;免疫杂交检测主动脉及细胞内NADPH氧化酶亚基gp91phox蛋白水平;Fenton反应Griess显色法测定血清及胸主动脉匀浆上清液活性氧(ROS)水平。I型胶原酶消化法培养人脐静脉内皮细胞(HUVECs),流式细胞术检测内皮细胞内ROS的水平,光泽精分析法测定NADPH氧化酶活性。采用干扰RNA和质粒转染的方法评价类视黄醇X受体α(RXRα)在Atorv抑制氧化应激中的作用。结果:(1)与C57组相比,ApoE-/-组小鼠胸主动脉斑块面积显著增加[(215.88±34.19)μm2vs 0μm2,P<0.01],2组间空腹血糖水平无显著差异,血清甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)、血清及胸主动脉ROS、胸主动脉gp91phox表达水平显著缩小(P<0.05);(2)与ApoE-/-组相比,STZ-ApoE-/-组胸主动脉斑块面积进一步增加[(314.13±35.72)μm2vs(215.88±34.19)μm2,P<0.05],血糖水平升高,血清TC、LDL-C、血清及胸主动脉ROS、胸主动脉gp91phox水平进一步增加(P<0.05);(3)与STZ-ApoE-/-组相比,STZ-ApoE-/-+Atorv组胸主动脉粥样斑块面积显著降低[(217.47±24.56)μm2vs(314.13±35.72)μm2,P<0.05],血糖、血清TG、HDL、TC、和LDL-C无显著变化,血清及胸主动脉ROS、胸主动脉gp91phox水平亦显著降低(P<0.05);(4)高糖(25 mmol/L)干预后,HUVECs内ROS含量、gp91phox蛋白水平及NADPH氧化酶活性明显增加(P<0.05),阿托伐他汀(10-8~10-6mol/L)显著降低高糖环境下HUVECs胞内ROS含量、gp91phox表达及NADPH氧化酶活性
AIM： To explore the effects of atorvastatin（ Atorv） on atherosclerosis in streptozotocin（ STZ）-induced diabetic apolipoprotein E knockout（ ApoE^-/-） mice with fat-rich diet and the possible mechanism. METHODS：C57 mice served as control. ApoE^-/-mice（ n = 34） fed with high-fat diet were randomly divided into ApoE^-/-group,STZ-ApoE^-/-group and STZ-ApoE^-/-＋ Atorv group. Intraperitoneal injection of streptozotocin was performed to create diabetic animal model. Blood glucose was determined by glucose oxidase method. Blood lipid levels were detected by enzymic method or selective homogeneous method. The plaque area in the thoracic aorta was measured by HE staining. The protein level of nicotinamide-adenine dinucleotide phosphate（ NADPH） oxidase subunit gp91^phoxin the thoracic aorta was determined by Western blotting. The levels of reactive oxygen species（ ROS） in blood and thoracic aorta homogenates were detected by Fenton reaction and Griess reagent. Human umbilical vein endothelial cells（ HUVECs） were isolated from healthy umbilical cords by collagenase I and cultured. ROS production was detected by flow cytometry. NADPH oxidase activity was measured using lucigenin assay. Effects of retinoid X receptor α（ RXRα） on inhibition of oxidative stress by atorvastatin were evaluated by RNA interference and plasmid transfection. RESULTS：（ 1） Compared with C57 group,the plaque areas of the thoracic aorta in ApoE^-/-group were increased. No difference of the fasting glucose between the 2groups was observed. The levels of triglyceride（ TG）,total cholesterol（ TC）,low-density lipoprotein cholesterol（ LDLC）,thoracic aorta gp91^phoxprotein and ROS in blood and thoracic aorta homogenates were higher in ApoE^-/-group than those in C57 group.（ 2） Compared with ApoE^-/-group,the plaque areas of the thoracic aorta in STZ-ApoE^-/-group were further enlarged [（ 314. 13 ± 35. 72） μm^2s（ 215. 88 ± 34. 19） μm^2P〈 0.05]. The levels of blood glucose,TG,TC and LDL-C,
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2014年第9期1537-1545,共9页
Chinese Journal of Pathophysiology
基金
国家自然科学基金青年基金资助项目(No.30900586)
