高脂高胆固醇饮食对3基因突变小鼠动脉粥样硬化性病变的影响

目的： 探讨高脂高胆固醇饮食性因素对3个脂代谢相关基因突变小鼠血脂代谢及动脉内膜损伤的影响。方法：分析高脂高胆固醇饮食喂养的3基因突变（ApoE-/-/LDLR-/-/Leprdb/db）小鼠血脂、血糖水平和主动脉内膜病变的特点。结果：高脂高胆固醇饮食喂养的3基因突变小鼠血浆总胆固醇（TC）、甘油三酯（TG）和血糖水平均显著高于普通饮食组。该饮食喂养2周和5周龄小鼠血浆TC、TG的浓度分别达(106.75±3.40) mmol/L和(9.12±1.35) mmol/L,高出普通饮食组4.33和2.36倍。主动脉内膜出现灶状内皮肿胀、脱落、单核/淋巴细胞黏附以及泡沫细胞形成。随喂养周数的增加出现内弹力板排列不整、部分断裂、内皮和平滑肌细胞内脂质沉积,并发生内膜增生型病变增多、范围扩大。血脂紊乱的加重与动脉内膜的损伤呈正相关。高脂高胆固醇饮食组小鼠的血脂紊乱和动脉内膜损伤均较普通饮食组严重,并伴有明显的肝细胞脂肪病变。结论：高脂高胆固醇饮食促进了3基因突变小鼠血脂代谢紊乱及主动脉内膜损伤的发生,提前并加重了动脉粥样硬化性病变的发生发展。     

脂代谢相关三基因突变小鼠肝组织基因表达差异研究

目的研究3个脂代谢相关基因联合突变小鼠与野生型小鼠肝脏基因表达差异及其与血脂代谢紊乱和动脉粥样硬化病变的关系。方法应用BiostarM-40S型小鼠cDNA表达谱芯片检测不同基因型小鼠肝脏基因表达的差异，并观察不同年龄小鼠血浆总胆固醇和甘油三脂的浓度以及主动脉内膜形态变化。结果在被测的4000条基因中，与野生型小鼠比较，5周龄三基因突变小鼠肝脏基因表达上调92条，下调105条，脂代谢相关基因中与胆固醇合成相关的基因表达下调，与甘油三脂代谢相关的肝脂肪酶基因表达水平上调。糖代谢、细胞骨架蛋白和免疫等相关的基因表达也有明显差异。5周龄的三基因突变小鼠血浆总胆固醇和甘油三脂水平明显高于野生型小鼠，伴有主动脉内膜损伤，并随年龄增长而加重。结论三基因突变导致肝脏中与脂类、糖类以及免疫等相关的多种基因表达改变，可能共同参与了血脂代谢紊乱和动脉粥样硬化的发生发展。     

ApoE^-/-母体孕期高脂高胆固醇饮食对子代成年鼠动脉粥样硬化的影响

目的研究母体孕期高脂高胆固醇饮食对子代成年鼠动脉粥样硬化的发生影响。方法ApoE^-/-孕鼠分为高脂高胆固醇饮食组（G组）和普通饮食组（P组）,G组雄性子代成年鼠分为高脂高胆固醇饮食组（G—G）和普通饮食组（G—P）,P组子代雄性成年鼠继续喂养普通饲料（P—P）,60d后检测子代成年鼠总胆固醇（TC）、甘油三酯（TG）、高密度脂蛋白（HDL）和低密度脂蛋白（LDL）浓度,HE染色观察血管形态学变化及动脉粥样硬化（atherosclerosis,AS）斑块形成情况。结果G—G组和G—P组的TC、TG、LDL和HDL浓度较P—P组明显升高,且与P—P组相比Tc浓度存在显著性差异（P〈0．05）,TG、LDL和HDL浓度存在极显著性差异（P〈0．01）;P—P组无AS斑块形成,G—G组与G．P组均有AS斑块形成,前者斑块形成率及斑块面积与血管管腔面积之比（PA／LA）都高于后者,两组PA／LA存在极显著性差异（P〈0．01）。结论母体孕期高脂高胆固醇饮食对子代成年鼠动脉粥样硬化的发生具有重要影响。     

高脂膳食对SR-AⅠ/Ⅱ基因敲除小鼠白色脂肪组织中UCP2 mRNA表达的影响

目的：观察SR-AⅠ/Ⅱ基因敲除（SR-AⅠ/Ⅱ-/-）小鼠白色脂肪组织中UCP2 mRNA的表达，探讨其与SR-AⅠ/Ⅱ-/-小鼠肥胖易感的关系。方法：应用荧光定量 RT-PCR检测小鼠附睾周白色脂肪组织中UCP2 mRNA的表达；图像分析法评估附睾周脂肪细胞形态学变化；酶法检测甘油三酯（TG）、总胆固醇（TC）、低密度脂蛋白胆固醇（LDL-C）血脂水平。结果：经12周高脂饲料喂养，SR-AⅠ/Ⅱ-/-小鼠的体重、血脂（TG、TC、LDL-C）水平、附睾周脂肪垫重量、脂肪细胞面积和直径均大于野生（SR-AⅠ/Ⅱ+/+）对照小鼠（P<0.01）；白色脂肪组织UCP2 mRNA表达量低于SR-AⅠ/Ⅱ+/+小鼠（P<0.01）。结论：SR-AⅠ/Ⅱ-/-小鼠对膳食诱导肥胖（diet-induced obesity, DIO）易感可能与其白色脂肪组织UCP2 mRNA表达异常有关。     

3,4,5,6-四羟基酮抑制ApoE基因缺陷小鼠动脉粥样硬化形成及机制

目的：研究3,4,5,6-四羟基（口山）酮抑制ApoE基因缺陷小鼠动脉粥样硬化（AS）形成的机制及其与内源性一氧化氮合酶（NOS）抑制剂非对称性二甲基精氨酸（ADMA）的关系.方法：32只小鼠分为4组（n=8）：正常对照组（C57BL/6J 小鼠,等体积溶媒灌胃）;模型组（ApoE基因缺陷小鼠,等体积溶媒灌胃）;低剂量（口山）酮组[ApoE基因缺陷小鼠,（口山）酮10 mg/（kg&#183;d）灌胃];高剂量（口山）酮组[ApoE基因缺陷小鼠,（口山）酮20 mg/（kg&#183;d）灌胃];检测主动脉脂质斑块面积、血脂、红细胞变形性和血浆ADMA的水平.结果：与模型组比较,（口山）酮可以显著降低血浆ADMA水平和斑块面积,降低血浆总胆固醇（TC）、血浆总甘油三酯（TG）、血浆低密度胆固醇（LDL-C）,显著升高血浆高密度胆固醇（HDL-C）和改善红细胞变形性.结论：3,4,5,6-四羟基（口山）酮具有抗AS作用,其作用机制与改善脂质代谢和降低血浆ADMA的水平有关.     

高脂高胆固醇膳食对小鼠PPARα基因表达及机体胆固醇水平的影响

目的:探讨高脂高胆固醇膳食对肝脏PPARα基因表达及机体胆固醇水平的影响。 方法:7-8周龄健康C57小鼠75只，随机分为5组，分别饲以相应配方饲料。 结果:与胆固醇膳食（Chol）相比，胆固醇＋多不饱和脂肪酸膳食（Chol＋PUFA）可增加(P<0.01)小鼠血清总胆固醇(TC)，增加的部分主要集中在HDL-C，降低肝脏胆固醇含量（P<0.01）；胆固醇＋单不饱和脂肪酸膳食（Chol＋MUFA）小鼠血清TC不变，肝脏胆固醇含量降低（P<0.01）；胆固醇＋饱和脂肪酸（Chol＋SFA）膳食可明显增加(P<0.01)小鼠血清TC，增加的部分主要集中在LDL-C，增加肝脏胆固醇含量（P<0.01）；此外，Chol+PUFA 膳食组小鼠肝脏PPARα mRNA和蛋白表达均高于Chol组（P<0.01），Chol+MUFA和Chol+SFA膳食组小鼠肝脏PPARα mRNA表达均明显低于Chol组（P<0.01）。 结论:在胆固醇基础上添加PUFA可诱导PPARα mRNA和蛋白的高表达,添加MUFA、SFA则抑制PPARα mRNA的表达；PPARα基因表达的改变可能进一步影响机体胆固醇水平。     

3,4,5,6-四羟基(口山)酮抑制ApoE基因缺陷小鼠动脉粥样硬化形成及机制

目的：研究3,4,5,6-四羟基口山酮抑制ApoE基因缺陷小鼠动脉粥样硬化(AS)形成的机制及其与内源性一氧化氮合酶(NOS)抑制剂非对称性二甲基精氨酸(ADMA)的关系。方法：32只小鼠分为4组(n=8):正常对照组(C57BL/6J小鼠，等体积溶媒灌胃)；模型组(ApoE基因缺陷小鼠，等体积溶媒灌胃)；低剂量口山酮组［ApoE基因缺陷小鼠，口山酮10mg/（kg·d）灌胃］；高剂量口山酮组［ApoE基因缺陷小鼠，口山酮20mg/（kg·d）灌胃］；检测主动脉脂质斑块面积、血脂、红细胞变形性和血浆ADMA的水平。结果：与模型组比较，口山酮可以显著降低血浆ADMA水平和斑块面积，降低血浆总胆固醇(TC)、血浆总甘油三酯(TG)、血浆低密度胆固醇(LDL-C)，显著升高血浆高密度胆固醇(HDL-C)和改善红细胞变形性。结论：3,4,5,6-四羟基口山酮具有抗AS作用, 其作用机制与改善脂质代谢和降低血浆ADMA的水平有关。     

高脂高糖食诱导的糖尿病小鼠肝脏清道夫受体B1的表达

目的：探讨糖尿病小鼠肝脏SR-B1表达变化，及其与血脂的关系。 方法： 进正常食C57BL/6J小鼠10只、进高脂高糖食8周小鼠5只、进高脂高糖食16周小鼠10只，测定血脂、血清胰岛素（INS）、空腹血糖（FBG），及肝脏SR-B1蛋白表达。 结果： ① 高脂高糖食16周的小鼠血清甘油三酯（TC）、FBG明显高于正常小鼠（P＜0.05)，其胰岛素水平与正常小鼠无显著差异（P＞0.05）。② 高脂高糖食小鼠肝脏SB-B1表达高于正常动物，16周强于8周的小鼠。 结论： 糖尿病小鼠肝脏SR-B1表达高于正常小鼠，血清HDL-C降低可能与肝脏的SR-B1蛋白表达增高有关。     

汉族人群脂联素基因-11377C/G多态性与子痫前期的相关性研究

目的探讨福建莆田地区汉族人群脂联素基因（APN）单核苷酸多态性（SNP）和子痫前期（pre-eclampsi-a）及其血脂水平的关联性。方法研究对象516例,包括对照组260例,子痫前期病例组256例。采用聚合酶链反应一限制性片段长度多态性（PCR-RFLP）法鉴定APN基因启动子-11377C/G单核苷酸多态性并测定血脂水平。结果 APN基因启动子-11377C/G位点G等位基因频率和GG＋CG基因型频率在子痫前期病例组明显升高,差异有统计学意义（P〈0.05）。子痫前期病例组内GG＋CG基因型患者血清甘油三酯（TG）、血浆总胆固醇（TC）和低密度脂蛋白（LDL）值均高于CC基因型（P〈0.05）。结论 APN基因启动子-11377C/G位点GG＋CG基因型与子痫前期及其血浆TG、TC和LDL水平升高关联,C-G多态性提高了子痫前期合并血脂代谢紊乱的风险性。     

丝瓜络对高脂血症小鼠LDL-R基因表达的影响

目的： 观察中药丝瓜络（RLF）对高脂血症小鼠低密度脂蛋白受体（LDL-R）基因表达的影响。方法：用高脂饲料喂养雄性昆明小鼠建立高脂血症模型,以血脂康作为阳性对照观察饲料中补充丝瓜络粉剂喂养对小鼠血清总胆固醇（TC）、低密度脂蛋白（LDL-C）的影响。按Trizol法提取小鼠肝脏总RNA,利用逆转录-聚合酶链反应(RT-PCR)测定LDL-R mRNA的表达,观察其在正常、高脂和给药3种条件下的差异。结果：（1）高脂组小鼠的血清TC和LDL-C分别为（5.71±0.82）和（3.99±1.12）mmol/L,显著高于正常对照组的TC（2.31±0.21）mmol/L和LDL-C（1.72±0.28）mmol/L（P＜0.01）,而高脂+丝瓜络组、高脂+血脂康组的TC分别为（3.65±0.28）mmol/L、（3.94±0.65）mmol/L和LDL-C分别为（2.74±0.54）mmol/L、（3.00±0.23）mmol/L,显著低于高脂组（P＜0.01）;（2）与正常对照组比较,高脂组小鼠肝脏LDL-R mRNA的表达减弱（P＜0.01）;与高脂组比较,高脂+丝瓜络组、高脂+血脂康组小鼠肝脏LDL-R mRNA的表达增强（P＜0.01）。结论： 丝瓜络对实验性高脂血症小鼠有明显的降血脂效应,且能使实验性高脂血症小鼠肝组织的LDL-R mRNA表达增强。     

ApoE/LDLR双基因突变小鼠肝脏差异表达蛋白组研究

目的：分析脂代谢相关双基因突变(apoE-/-/LDLR-/-)小鼠肝脏蛋白质表达特点，研究差异表达蛋白与基因突变小鼠血脂代谢紊乱和动脉粥样硬化的关系。 方法: 应用双向电泳及质谱技术对5周龄双基因突变和野生型小鼠肝组织差异蛋白进行比较研究。结果: 双基因突变和野生型小鼠肝脏中分别检测到（928±15）和（1 017±50）个蛋白点（n=3），两者之间的平均匹配率分别为78.7%和83.2%。双基因突变小鼠有108个蛋白点未能与野生型小鼠匹配，相差5倍以上的上调点和下调点分别为10个和45个，取其中6个点做质谱分析，鉴定为endoplasmin precursor,acidic leucin-rich nuclear phosphoprotein 32 family member A,serotransferrin precursor,stress-70 protein precursor,fibronectin precursor,complement C3 precursor,fibrinogen gamma polypeptide 7种蛋白质。 结论: 双基因突变小鼠与野生型小鼠的肝脏蛋白表达谱有明显差异，推测这些蛋白在脂代谢相关双基因突变引发的小鼠血脂代谢紊乱在动脉粥样硬化的发生发展中可能发挥重要作用。     

Blockade of IL-6 signaling exacerbates liver injury and suppresses antiapoptotic gene expression in methionine choline-deficient diet-fed db/db mice

Our previous study revealed that blockade of interleukin-6 (IL-6)-STAT3 signaling ameliorated liver injury, although hepatic STAT3(-/-) or GP130(-/-) mice have been reported to develop severe liver injury, in a murine methionine choline deficient (MCD) diet-induced model of non-alcoholic steatohepatitis (NASH). In this study, to determine whether profound blockade of IL-6-STAT3 signaling may still ameliorate liver injury, we studied db/db mice, which have impaired leptin-mediated STAT3 activation, using the MCD diet-induced NASH model. Male lean and db/db mice (6 weeks old) were fed either control chow or an MCD diet for 8 or 12 weeks. Half of the mice were treated with 15 mg/kg rat anti-mouse IL-6 receptor neutralizing antibody (MR16-1) intraperitoneally twice weekly, the remainder were injected with 15 mg/kg rat IgG as a control. Hepatic steatosis, injury, fibrosis, markers of lipid peroxidation/oxidant stress and antiapoptotic gene expression were evaluated. Plasma IL-6 levels were elevated in all groups of db/db mice. Although hepatic IL-6/ GP130 signaling was activated in chow-fed db/db mice, this was suppressed in MCD diet-fed db/db mice, accompanied by downregulation of hepatic IL-6 receptor and GP130 mRNA expression. MR16-1 treatment of MCD diet-fed db/db mice further repressed STAT3 activities and expression of STAT3-related antiapoptotic genes, such as Bcl-2 and Ref-1, but increased plasma-free fatty acid and hepatic markers of lipid peroxidation/oxidant stress, leading to increased liver injury, hepatocyte apoptosis and liver fibrosis. Although 'moderate' blockade of enhanced IL-6-STAT3 signaling may be beneficial in NASH, as we reported previously, these findings demonstrate that a profound defect in STAT3 activation is detrimental in terms of liver injury, hepatocyte apoptosis and liver fibrosis, indicating the hepato-protective role of IL-6 signaling in this severe NASH model.     

甜菜碱对老龄db/db小鼠脂肪性肝损害的影响

 目的：通过高脂饮食诱发db/db小鼠非酒精性脂肪性肝病（NAFLD）模型,探索甜菜碱对遗传性小鼠脂肪肝脂质代谢的影响。方法：50只7月龄db/db鼠随机分为低、中、高剂量组、生理盐水对照组和阳性药物组。所有小鼠均饲以高脂饲料, 以诱发NAFLD 模型。 小鼠分别以200 mg/kg(低剂量组)、400 mg/kg(中剂量组)和800 mg/kg(高剂量组)甜菜碱溶液灌胃,连续6周。测定血清丙氨酸氨基转移酶(ALT)、甘油三酯(TG)、总胆固醇（TC）和低密度脂蛋白(LDL)水平,并行葡萄糖耐量测定和肝组织病理学观察。结果：甜菜碱可显著降低血清ALT、TC和LDL的水平(P<0.05或P<0.01)。组织学结果表明甜菜碱可显著减少小鼠肝细胞的脂肪样变性。结论: 甜菜碱可以显著改善老龄db/db小鼠的脂类代谢紊乱和肝功能,明显降低脂肪在肝细胞中的积蓄。     

apoE/LDLR双基因突变小鼠肝脏脂代谢相关基因的表达研究

目的探讨载脂蛋白E和低密度脂蛋白受体双基因缺失（apoE^-/-/LDLR^-/-）小鼠肝脏脂代谢相关基因表达特征与动脉粥样硬化早期病变的关系及发生机制。方法应用RT-PCR技术检测apoE^-/-／LDLR^-/-与野生型小鼠肝脏脂代谢相关基因表达差异,并进行血生化指标检测及主动脉形态学观察。结果所检测的11个脂代谢相关基因中,apoE^-/-／LDLR^-/-小鼠与野生型小鼠相比,载脂蛋白B100和脂肪酸转运体（FAT／CD36）的mRNA水平从14天龄至3月龄均升高。载脂蛋白AⅣ、载脂蛋白AⅤmRNA水平分别于14天龄和3月龄时明显降低。载脂蛋白AⅠ、载脂蛋白F、过氧化物增殖物激活型受体α、肝X受体α、血管生成素样蛋白3、酰基辅酶A氧化酶1及肉碱棕榈酰转移酶ImRNA表达水平无明显差异。血清总胆固醇、总甘油三酯和低密度脂蛋白胆固醇水平分别高于同龄野生型小鼠约7、2和30倍。随年龄增长apoE^-/-／LDLR^-/-小鼠主动脉内膜出现典型的动脉粥样硬化早期病变。结论上述脂代谢相关基因表达变化表明它们在动脉粥样硬化发生发展过程中起重要作用。     

Elevation of blood lipids in hepatocyte-specific fatty acid transport 4-deficient mice fed with high glucose diets

Fatty acid transport protein4 (FATP4) is upregulated in acquired and central obesity and its polymorphisms are associated with blood lipids and insulin resistance. Patients with FATP4 mutations and mice with global FATP4 deletion exhibit skin abnormalities characterized as ischthyosis prematurity syndrome (IPS). Cumulating data have shown that an absence of FATP4 increases the levels of cellular triglycerides (TG). However, FATP4 role and consequent lipid and TG metabolism in the hepatocyte is still elusive. Here, hepatocyte-specific FATP4 deficient (Fatp4^L?/?) mice were generated. When fed with chow, these mutant mice displayed no phenotypes regarding blood lipids. However when fed low-fat/high-sugar (HS) or high-fat/high-sugar (HFS) for 12?weeks, Fatp4^L?/? mice showed a significant increase of plasma TG, free fatty acids and glycerol when compared with diet-fed control mice. Interestingly, Fatp4^L?/? mice under HS diet had lower body and liver weights and they were not protected from HFS-induced body weight gain and hepatic steatosis. Male mutant mice were more sensitive to HFS diet than female mutant mice. Glucose intolerance was observed only in female Fatp4^L?/? mice fed with HS diet. Lipidomics analyses revealed that hepatic phospholipids were not disturbed in mutant mice under both diets. Thus, hepatic FATP4 deletion rendered an increase of blood lipids including glycerol indicating a preferential fatty-acid channeling to TG pools that are specifically available for lipolysis. Our results imply a possible risk of hyperlipidemia as a result of abnormal metabolism in liver in IPS patients with FATP4 mutations who consume high-sugar diets.     

Microarray profiling of gene expression patterns in glomerular cells of astaxanthin-treated diabetic mice: a nutrigenomic approach

We have demonstrated that astaxanthin reduces glomerular oxidative stress as well as inhibits the increase in urinary albumin in diabetic db/db mice. The aim of the present study was to determine the gene expression patterns in the glomerular cells of the diabetic mouse kidney, and to investigate the effects of astaxanthin on the expression of these genes using a high-density DNA microarray. The diet administered to the astaxanthin-supplementation group was prepared by mixing a control powder with astaxanthin at a concentration of 0.02%. Glomerular cells were obtained from the kidneys of mice by laser capture microdissection. Preparation of cRNA and target hybridization were performed according to the Affymetrix GeneChip eukaryotic small sample target labeling assay protocol. The gene expression profile was evaluated by the mouse expression set 430A GeneChip. Array data analysis was carried out using Affymetrix GeneChip operating and Ingenuity Pathway analysis software. Comparison between diabetic db/db and non-diabetic db/m mice revealed that 779 probes (3.1%) were significantly affected, i.e. 550 probes were up-regulated, and 229 probes were down-regulated, both at levels of >/=1.5-fold in the diabetic mice. Ingenuity signal analysis of 550 up-regulated probes revealed the mitochondrial oxidative phosphorylation pathway as the most significantly affected caronical pathway. The affected genes were associated with complexes I, III, and IV located on the mitochondrial inner membrane, and the expression levels of these genes were decreased in mice treated with astaxanthin as compared to the levels in the control mice. In addition, the expression of many genes associated with oxidative stress, collagen synthesis, and transforming growth factor-beta signaling was enhanced in the diabetic mice, and this enhancement was slightly inhibited in the astaxanthin-treated mice. In conclusion, this genome-wide nutrigenomics approach provided insight into genes and putative genetic pathways that are thought to be affected by stimulation by high-glucose concentrations. In addition, the present approach may help us gain a better understanding of the genes and pathways involved in the anti-diabetic mechanism of astaxanthin.