Tag single nucleotide polymorphism rs1532624 located in cholesteryl ester transfer protein gene is associated with atherosclerosis cerebral ischemia

Objective: To investigate the relationship between polymorphisms of rs1532624 and rs289741 loci in cholesteryl ester transfer protein(CETP) genes and atherosclerotic cerebral infarction(ACI). Methods: The CETP gene rs1532624 and rs289741 in 95 patients with ACI and 177 healthy subjects were genotyped by Mass ARRAY mass spectrometry. Each locus genotype and allele frequency distributions were compared. Results: The difference of allele frequency distribution between the rs1532624(χ~2=1.723, P=0.189) and rs289741(χ~2=2.466, P=0.116) were not statistically significant. The frequency distribution of rs1532624 genotype between the cerebral infarction group and healthy control group was statistically significant(χ~2=7.096, P=0.029), while rs289741 genotype frequency distribution between the two groups was not statistically significant(χ~2=2.906, P=0.234). Conclusion: ACI have a positive correlation with rs1532624 polymorphism, and AA genotype may be susceptible factors of ACI.

Qualitative Distribution of Endogenous Cholesteryl Esters in Plasma of Humans and Three Rodent Species Using Stepwise UPLC-Q-Exactive-MS

Objective:Cholesteryl esters(CEs)are composed of various fatty acyl chains attached to the hydroxyl groups of cholesterol,and abnormalities in their metabolism are related to many diseases.This study aimed to develop an ultrahigh-performance liquid chromatography-quadrupole exactive mass spectrometry(UPLC-Q-Exactive MS)method to identify the CEs in plasma.Methods:First,the MS fragmentation patterns were investigated using seven commercial CE standards.Then,the CEs in plasma were characterized through the accurate mass data of precursor ions and characteristic product ions.A strategy of step-by-step m/z scans in a narrow range was proposed to identify more trace CEs by the full-scan data-dependent MS/MS(ddMS2)mode.Results:A total of 50 CE species consisting of 55 regioisomers were identified in human plasma.Among them,two species were reported for the first time.Conclusion:This study is the most comprehensive identification of CE species in human plasma to date.These results will contribute to the in-depth profiling of CEs in human plasma and provide guidance for animal model selection when studying lipid-related diseases.

SPECTROSCOPIC DETERMINATION OF THE AVERAGE NUMBER OF COAGGREGATES (Nco) OF CHOLESTERYL ESTERS AND THE FLUORESCENCE PROBE

Average aggregate number of coaggregates(N_co)of CE-n or BL-n and the fluoresc- ence probe(Np-16)have been determined by using time-resolved fluorescence spectroscopy. Chain-length,hydroxy-group and chain-foldability effects on the N_co have been discussed.

Synthesis and Characterization of Mesomorphic Behaviour of New Mesogenic Compounds Incorporating Cholesteryl Ester Moiety Connected to 1,3,4-Oxadiazole

The synthesis and characterization of new series of liquid crystals containing cholesteryl moiety connected via an ester linkage to 1,3,4-oxadiazole are reported. The molecular structures of the intermediates and target compounds were confirmed by elemental analyses and FT-IR, ^1H NMR and mass spectrometry. The mesomorphic behaviors were investigated by polarizing optical microscopy （POM） attached with hot stage and differential scanning calorimetry （DSC）. Enantiotropic smectic 1, smectic A, blue phase and cholesteric mesophases are exhibited by the newly synthesized compounds.

High level of serum cholesteryl ester transfer protein inactive hepatitis C virus infection

AIM: To determine the significance of cholesteryl ester transfer protein(CETP) in lipoprotein abnormalities in chronic hepatitis C virus(HCV) infection.METHODS: We evaluated the significance of the serum concentration of CETP in 110 Japanese patients with chronic HCV infection. Fifty-five patients had active HCV infection, and HCV eradication had been achieved in 55. The role of CETP in serum lipoprotein abnormalities, specifically, in triglyceride(TG) concentrations in the four major classes of lipoproteins, was investigated using Pearson correlations in conjunction with multiple regression analysis and compared them between those with active HCV infection and those in whom eradication had been achieved. RESULTS: The serum CETP levels of patients with active HCV infection were significantly higher than those of patients in whom HCV eradication was achieved(mean ± SD, 2.84 ± 0.69 μg/m L vs 2.40 ± 1.00 μg/m L, P = 0.008). In multiple regression analysis, HCV infection status(active or eradicated) was an independent factor significantly associated with the serum CETP level. TG concentrations in low-density lipoprotein(mean ± SD, 36.25 ± 15.28 μg/m L vs 28.14 ± 9.94 μg/m L, P = 0.001) and high-density lipoprotein(HDL)(mean ± SD, 25.9 ± 7.34 μg/m L vs 17.17 ± 4.82 μg/m L, P < 0.001) were significantly higher in patientswith active HCV infection than in those in whom HCV eradication was achieved. The CETP level was strongly correlated with HDL-TG in patients with active HCV infection(R = 0.557, P < 0.001), whereas CETP was not correlated with HDL-TG in patients in whom HCV eradication was achieved(R =-0.079, P = 0.56). CONCLUSION: Our results indicate that CETP plays a role in abnormalities of lipoprotein metabolism in patients with chronic HCV infection.

Pharmacophore-based design,synthesis,and biological evaluation of novel 3-((3,4-dichlorophenyl)(4-substituted benzyl)amino)propanamides as cholesteryl ester transfer protein(CETP)inhibitors

Cholesteryl ester transfer protein （CETP） is a plasma glycoprotein that plays an important role in decreasing high-density lipoprotein cholesterol （HDL-C） levels and increasing low-density lipoprotein cholesterol （LDL-C） levels. Inhibition of CETP may be a new therapy for treating atherosclerosis. Herein, we report the development of a ligand-based pharmacophore model and pharmacophore-based virtual screening of the ZINC/big-n-greasy database, leading to the identification of compound H-10 as a potential CETP inhibitor in vitro. Based on H-10, a series of 3-（（3,4-dichlorophenyl）（4-substituted benzyl）amino） propanamides were designed, synthesized, and evaluated against CETP. Compound 41 was found to have the best activity, resulting in 85.0% inhibition of CETP at l0 μmol/L.

Association between TaqIB polymorphism of cholesteryl ester transfer protein and coronary artery disease in the Chinese population

Objective: To assess whether the TaqIB polymorphism of cholesteryl ester transfer protein (CETP) is associated with coronary artery disease (CAD) in Chinese population, we performed a meta-analysis in this paper. Methods: We searched PubMed, Embase, the Science Citation Index (SCI), the China Biological Medicine database (CBM), the China National Knowledge Infrastructure (CNKI), and the Wanfang database for relevant articles. Data were extracted, and pooled odds ratios (ORs) and 95% confidence intervals (CIs) were calculated. Results: The lit- erature search yielded 448 studies, in which 10 case-control studies including 1 694 cases and 1 456 controls matched the selection criteria. The combined B1 and B2 allele frequencies were 0.587 and 0.413, respectively. The pooled OR was 1.10 (95% CI, 0.89-1.34) for comparing the B1B1 or B1B2 carriers with B2B2 carriers, and was 1.27 (95% CI, 1.09-1.49) in the B1B1 carriers versus B2B2 or B1B2 carriers. Conclusions: In the present study, the TaqIB poly- morphism of CETP was found to be associated with CAD in the Chinese population.

Taq1B polymorphism and plasma activity of cholesteryl ester transfer protein and restenosis after coronary stent placement

Cholesteryl ester transfer protein （CETP）, an important regulatory factor in reverse cholesterol transport, has been investigated in relation to plasma HDL cholesterol, which is associated with restenosis after percutaneous transluminal coronary angioplasty （PTCA） and stent implantation. We examined the effect of plasma CETP activity and the Taq1B polymorphism on restenosis in Chinese HaM population. Methods and Results Our study included 228 patients with symptomatic coronary artery disease （CAD）. The genotype of the subjects for Taq1B polymorphism of CETP was analyzed by using polymerase chain reaction-restriction fragment length poly- morphism （PCR-RFLP）. In-stent restenosis （ISR） was observed in 93 patients （40.8 %）. Baseline clinical, angiographic,and procedural characteristics data were not significantly different in patients with and without restenosis,except previous history of myocardial infarction （MI）, left ventricular ejection fraction （LVEF）, statin medication, and stent type. Taq1B polymorphism and plasma activity of CETP were not associated with the incidence of ISR （P = 0.68, P = 0.30, respectively）. Linear regression analysis revealed that the risk of ISR was statistically significant correlation with stent type, LVEF, statin medication, and lesion length （P 〈 0.05）. Conclusions Useful markers for risk of ISR were stent type, LVEF, statin medication and lesion length but not the Taq1B polymorphism and plasma activity of CETP.

中国健康人血清胆固醇酯转运蛋白水平

为研究中国健康人血清胆固醇酯转运蛋白水平和分布特征 ,采用“Sandwich”酶免疫法测定了 112 8例健康人群血清胆固醇酯转运蛋白水平 ,结果为 1.84± 1.5 5mg/L ,胆固醇酯转运蛋白水平倾向于低浓度的偏态分布 ,女性胆固醇酯转运蛋白水平明显高于男性 (分别为 2 .40± 1.6 5mg/L和 1.49± 1.37mg/L ,P <0 .0 0 1)。胆固醇酯转运蛋白水平与年龄之间呈显著的负相关 (r=- 0 .19,P <0 .0 0 1) ,与其他各项血脂浓度间均无相关性。

豚鼠早期动脉粥样硬化模型的建立、机制研究及与大鼠模型的比较

目的建立豚鼠早期动脉粥样硬化模型,探讨模型形成机制,并与大鼠模型进行比较,为豚鼠模型优势提供科学依据。方法应用高脂饲料诱导法,观察豚鼠和大鼠是否可形成早期动脉粥样硬化模型,并应用免疫组化和酶联免疫分析技术探讨模型形成机制。结果豚鼠模型组摄入高脂饲料6周后主动脉内膜-中膜明显增厚、内膜单核细胞、巨噬细胞浸润与聚集增多,同时比例为0.40的动物动脉内膜表层进一步发展形成脂纹脂斑病变。而大鼠模型组未见类似病变。机制研究表明,血清脂蛋白(a)[Lipoprotein(a),LP(a)]、胆固醇酯转运蛋白(Cholesteryl ester transfer protein,CETP)、氧化型低密度脂蛋白(ox-LDL)浓度,肝脏脂酰辅酶A:胆固醇酰基转移酶(Acyl-CoA:cholesterol acyltransferase,ACAT)活性和主动脉增殖细胞核抗原(Proliferating cell nu-clear antigen,PCNA)、分化抗原簇-36(Cluster of differentiation36,CD36)和血管细胞黏附分子(Vascular cell adhesion mole-cule,VCAM-1)表达的变化是动脉病理改变的重要机制。结论豚鼠可能是一种模拟早期动脉粥样硬化的适宜动物模型。

脂肪分化相关蛋白通过酰基辅酶A∶胆固醇酰基转移酶1促进巨噬细胞脂质蓄积

目的观察高表达脂肪分化相关蛋白对酰基辅酶A∶胆固醇酰基转移酶1表达及脂质蓄积的影响。方法构建表达载体pcDNA3.1-HA-脂肪分化相关蛋白,使用THP-1巨噬细胞,通过瞬时转染使之高表达脂肪分化相关蛋白,依次用氧化型低密度脂蛋白和(或)丙泮尼地处理。逆转录聚合酶链反应和Western blot检测酰基辅酶A∶胆固醇酰基转移酶1及脂肪分化相关蛋白的表达,油红O染色和高效液相色谱检测细胞内脂质的蓄积。结果随着氧化型低密度脂蛋白浓度的增加,巨噬细胞脂肪分化相关蛋白及酰基辅酶A∶胆固醇酰基转移酶1表达明显增强,两者呈伴行关系。丙泮尼地能抑制酰基辅酶A∶胆固醇酰基转移酶1表达上调,且随处理时间延长,其表达逐渐减少,并且能减少细胞内脂滴生成。与对照组相比,瞬时转染pcDNA3.1-HA-脂肪分化相关蛋白能使脂肪分化相关蛋白表达明显升高。高表达脂肪分化相关蛋白的巨噬细胞能使酰基辅酶A∶胆固醇酰基转移酶1表达增加,促进细胞内胆固醇酯蓄积,并协同增强氧化型低密度脂蛋白的作用。加入丙泮尼地后,高表达脂肪分化相关蛋白的作用被减弱。结论高表达脂肪分化相关蛋白能上调THP-1巨噬细胞酰基辅酶A∶胆固醇酰基转移酶1表达,促进细胞内脂质蓄积。脂肪分化相关蛋白可能通过酰基辅酶A∶胆固醇酰基转移酶1促进细胞内胆固醇酯的蓄积。

普伐他汀对泡沫细胞内胆固醇酯的影响及与小凹蛋白-1的关系

本文旨在观察普伐他汀对鼠源巨噬细胞性泡沫细胞内胆固醇酯含量的影响,探讨此作用与小凹蛋白-1的关系。采用体外培养的鼠源性巨噬细胞株作为研究对象,加入氧化低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)使其形成泡沫细胞,运用高效液相色谱测定细胞内胆固醇酯的改变,同时运用Western blot检测细胞中小凹蛋白-1的表达,并观察普伐他汀对细胞内胆固醇酯和小凹蛋白-1影响的量效和时效关系。结果显示:普伐他汀可明显降低泡沫细胞内的胆固醇酯与总胆固醇的比值,且在一定范围内呈剂量依赖性和时间依赖性。在泡沫细胞中加入普伐他汀后能够促进小凹蛋白-1的表达,呈剂量依赖性和时间依赖性。上述结果提示普伐他汀通过降低细胞内胆固醇酯的含量,减轻细胞泡沫化程度。普伐他汀的这一作用可能与促进小凹蛋白-1表达上调有关。

氧化低密度脂蛋白诱导主动脉平滑肌细胞胆固醇酯聚集和凋亡(英文)

细胞内胆固醇代谢的失衡和细胞凋亡都与动脉粥样硬化的发生有关 .为了研究两者之间的关系 ,我们把猪的主动脉平滑肌细胞与 15mg/L氧化低密度脂蛋白共同孵育 72h ,发现细胞内胆固醇酯与总胆固醇的比值由 2 6 2 %增加到 6 4 1% ,并且细胞内胆固醇酯的积聚有剂量依赖关系 ,表明细胞已经转化为平滑肌源性的泡沫细胞 .另外 ,使用荧光显微镜、激光共聚焦显微镜和流式细胞仪分别发现 ,与氧化低密度脂蛋白共孵育的细胞有典型的凋亡形态改变 .从实验可以推测 ,由氧化低密度脂蛋白诱导的平滑肌细胞凋亡 ,除了低密度脂蛋白氧化的因素外 ,也可能与细胞内胆固醇酯与总胆固醇的比值升高有关 .

CETP基因TaqⅠB位点多态性与高胆固醇血症的关系

目的探讨胆固醇酯转运蛋白CETP基因TaqⅠB位点多态性与汉族人群高胆固醇血症的关系。方法应用聚合酶链技术-限制性片段长度多态性(PCR-RFLP)技术检测汉族人群464例高胆固醇血症(高胆固醇血症组)、852例边缘高胆固醇血症(边缘高胆固醇血症组)及1 309例正常血脂(正常对照组)人群CETP基因内含子1的TaqⅠB位点多态性。结果按TaqⅠ酶切位点存在与否分为B1B1、B1B2、B2B2 3种基因型和B1、B2 2种等位基因。高胆固醇血症组较边缘高胆固醇血症组、正常对照组LDL-C、CETP升高,HDL-C降低(P<0.05)。CETP基因TaqⅠB多态性基因型及等位基因频率在3组人群中的分布差异有统计学意义,高胆固醇血症组和边缘高胆固醇血症组的B1B1基因型及B1等位基因频率显著高于正常对照组(P<0.05)。CETP基因TaqⅠB多态性B1B1基因型者血清TC、LDL-C和CETP水平明显高于B1B2基因型和B2B2基因型(P<0.05)。多因素logistic回归分析显示CETP基因TaqⅠB B1B1基因型是高胆固醇血症的独立危险因素。结论胆固醇酯转运蛋白基因TaqIB位点存在基因多态性,B1B1基因型及B1等位基因与高胆固醇血症有着密切的关系,可能是汉族人群高胆固醇血症的原因之一。

冠心病患者血浆胆固醇酯转运蛋白水平与高密度脂蛋白亚类组成的关系

目的探讨冠心病患者胆固醇酯转运蛋白(CETP)水平与血浆高密度脂蛋白(HDL)亚类组成的关系。方法收集冠心病患者血浆116例和健康对照组血浆87例,采用双向电泳-免疫印迹检测法分析其HDL亚类浓度,用酶联免疫法测定CETP浓度。按CETP浓度均值加或减去一个标准差作为分割点,将冠心病患者分为3组:低CETP组(CETP≤0.69 mg/L)、中CETP组(0.69<CETP<1.59 mg/L)、高CETP组(CETP≥1.59 mg/L)。结果随着CETP浓度的升高,冠心病患者血浆总胆固醇、甘油三酯、低密度脂蛋白胆固醇水平呈增加趋势,高密度脂蛋白胆固醇、载脂蛋白A1水平和载脂蛋白A1/B100比值呈减少趋势。与低CETP组相比,中CETP组和高CETP组preβ_1-HDL及HDL_(3c)含量增加,而HDL_(2b)含量降低(P<0.05或P<0.001)。直线相关和多元回归分析中发现,血浆CETP水平与preβ_1-HDL浓度呈正相关,与HDL_(2b)浓度呈负相关。结论随着血浆CETP水平的升高HDL颗粒有变小的趋势,CETP可能阻碍了HDL的成熟代谢。

CETP基因TaqI B多态性与瑞苏伐他汀对糖耐量异常合并冠心病患者疗效的相关性

目的:比较血糖异常合并冠心病组瑞舒伐他汀治疗前后的血脂指标和冠状动脉粥样硬化斑块变化间的关系。方法:血糖异常合并冠心病患者196例及血糖正常160例给予瑞苏伐他汀治疗,分析病例组治疗前后冠脉斑块的血管内超声(intravascular ultrasound,IVUS)结果以及血脂变化情况,用飞行时间质谱检测研究对象血标本的CETP基因TaqI B单核苷酸多态性(SNPs)位点。结果:CETP基因TaqI B B1B1、B1B2、B2B基因型在病例组的分布频率分别为35.7%、48.0%,16.3%;对照组为31.3%、53.1%、15.6%。HDL-C及MLA在瑞苏伐他汀治疗后呈上升趋势,LDL-C、TG、TCH、Lpa、PA、EEMA及PB治疗后呈下降趋势。结论:CETP基因Taql B多态性与瑞苏伐他汀对冠心病患者的血脂调节以及血管斑块退缩作用可能有相关性。

CETP基因启动子区多态与他汀类药物调血脂效果的关联分析

目的探讨胆固醇酯转运蛋白基因启动子区-69G/A、-629C/A、-971G/A和-1337C/T单核苷酸多态性与他汀类药物调血脂效果的关系。方法采用聚合酶链反应-限制性片段长度多态方法检测207例经辛伐他汀调脂治疗的血脂异常患者胆固醇酯转运蛋白基因4个多态性位点的基因型频率分布情况,并比较各项血脂水平的改变量在各多态位点基因型之间的差异。结果 -69G/A多态未检出A等位基因;辛伐他汀治疗后,总胆固醇和低密度脂蛋白胆固醇水平的改变量在-629C/A多态各基因型组间差异均有显著性(P=0.025;P=0.000),-629AA基因型个体的总胆固醇水平下降幅度较CC基因型个体要高出0.495 mmol/L(P=0.020),-629AA和CA基因型个体中的低密度脂蛋白胆固醇水平下降幅度比CC基因型个体分别高出0.577 mmol/L和0.352 mmol/L(P=0.000,P=0.012);高密度脂蛋白胆固醇水平的改变量在-1337C/T多态各基因型组间差异有显著性(F=3.064,P=0.044),高密度脂蛋白胆固醇水平升高的幅度在-1337TT、TC和CC基因型组间有逐渐增大的趋势;各项血脂水平的改变量在-971G/A多态3种基因型组间的差异无统计学意义。结论 -629C/A多态和-1337C/T多态与他汀类药物调血脂效果有关联。

冠心病患者血清磷脂转运蛋白及胆固醇脂转运蛋白水平与中医证型相关性研究

目的探讨冠心病患者血清磷脂转运蛋白(phospholipid transfer protein,PLTP)、胆固醇酯转运蛋白(cholesteryl ester transfer protein,CETP)水平与中医病证的相关性,为冠心病证候客观化和开发临床治疗药物探索新的途径。方法选择经冠状动脉造影确诊为冠心病的患者201例,采用国内外公认标准,构建证候四诊合参数据库,进行中医辨证。随机选取健康体检者20名为健康对照组。取静脉血分离血清,分别以酶联免疫法测定血清PLTP、CETP表达;采用乙酰丙酮显色法测定甘油三酯(triglyceride,TG)含量;采用沉淀漂浮酶联法测定总胆固醇(total cholesterol,TC)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-C)、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-C)含量。结果各证型冠心病患者血清PLTP、CETP、TC明显高于健康对照组,差异有统计学意义(P<0.05,P<0.01);各证型组血清TG、HDL-C、LDL-C与健康对照组比较,差异有统计学意义(P<0.05,P<0.01)。标实证组各证型血清PLTP与本虚组各证型比较,差异有统计学意义(P<0.05)。标实证组各证型血清TG、HDL-C、LDL-C与本虚组各证型比较,差异有统计学意义(P<0.05、P<0.01)。结论标实证冠心病患者血清PLTP表达明显升高。标实证冠心病患者血清脂质改变较本虚证更为显著。

人胆固醇酯水解酶在RAW 264.7巨噬细胞内瞬时表达对胆固醇代谢的影响

目的:以RAW264.7小鼠单核巨噬细胞为研究对象,探讨人胆固醇酯水解酶(Human cholesteryl ester hydrolase,hCEH)在鼠巨噬细胞内瞬时表达,对细胞内胆固醇代谢产生的影响。方法:利用FuGENE HD转染试剂将pEGFP-N1-hCEH质粒,转染RAW264.7小鼠单核巨噬细胞。24 h后将巨噬细胞与氧化型低密度脂蛋白(Oxidized low density lipoprotein,oX-LDL)(100 mg/L),共同孵育48 h。荧光显微镜观察及Western blot检测细胞内CEH的表达,油红O染色观察细胞内脂滴堆积情况,高效液相色谱检测细胞内游离胆固醇(Free cholesterol,FC)和胆固醇酯(Cholesterol esters,CE)含量。结果:转染72 h后,荧光细胞数最多,转染率接近40%。Western blot检测显示,hCEH蛋白在鼠巨噬细胞内大量表达并部分抑制鼠胆固醇酯水解酶(Cholesteryl ester hydrolase,CEH)表达。转染hCEH组与ox-LDL诱导组相比,油红O染色阳性细胞数显著减少,细胞内总胆固醇、游离胆固醇和胆固醇酯含量明显下降。结论:hCEH基因在RAW264.7小鼠单核巨噬细胞内成功表达,减少细胞内胆固醇水平,抑制泡沫细胞的形成。

高效液相色谱分析氧化型低密度脂蛋白处理的U937细胞胞内胆固醇及胆固醇酯

为找到一种判断泡沫细胞与非泡沫细胞的简便而较为实用的方法，本文采用正己烷-异丙醇从U937单核细胞内提取胆固醇，并用醇溶性氢氧化钾除去其中的甘油三酯成分，用三氯乙酸去其中的蛋白成分。以异丙醇：正己烷：乙腈为溶剂洗脱系统，采用疏水性较弱的核酸分离柱，进行高效液相色谱分析，分离纯化胆固醇和胆固醇酯成分，并采用L－B（LiebermanBurchard）显色法结合波谱分析法对所胆固醇峰进行鉴定。结果表明胆固醇含量在0．05～1g／L之间与其峰面积有较好的线性关系，其最低检出量为4mg／g细胞蛋白（100mg细胞蛋白相当于1×108个细胞），通过定性与定量分析，结果表明，U937单核细胞株经氧化型低密度脂蛋白处理48h后，其胞内的胆固醇酯占胞内总胆固醇60％以上。