Association between apolipoprotein E gene polymorphism and nonalcoholic fatty liver disease in Southern China: A case‐control study

BackgroundApolipoprotein E (ApoE) polymorphisms have been reported to be associated with nonalcoholic fatty liver disease (NAFLD), but the conclusions of studies are inconsistent in different regions. The present study aims to investigate the role of ApoE genotypes on NAFLD in southern China.MethodsA total of 1064 subjects including 372 NAFLD patients and 692 controls who attended Meizhou People''s Hospital located in southern China from March 1, 2016 to April 30, 2020 were enrolled in this study. The ApoE genotypes were detected and the laboratory parameters were examined.ResultsSignificant differences were observed between NAFLD patients and controls in the prevalence of ε3/ε3 (p < 0.001) and ε3/ε4 (p = 0.004). NAFLD patients presented higher frequency of ε4 allele than controls (p = 0.013). Logistic regression analysis suggested that ε3/ε3 was an independent risk factor (OR: 1.435, 95% CI: 1.084–1.891, p = 0.010), while ε3/ε4 was an independent protective factor (OR: 0.578, 95% CI: 0.404–0.828, p = 0.003) for development of NAFLD. In addition, allele ε4 showed a protective effect on NAFLD with an adjusted OR of 0.588 (95% CI: 0.420–0.824, p = 0.002).ConclusionOur results suggested that ApoE genotype was associated with the development of NAFLD in the population of southern China. Individuals carrying ε3/ε3 were at higher risk of NAFLD, while those carrying ε3/ε4 were at lower risk of NAFLD.     

载脂蛋白E对结直肠癌细胞侵袭迁移能力的影响

目的 观察载脂蛋白E（apolipoprotein E, ApoE）对结直肠癌细胞侵袭迁移能力的影响。方法 在结直肠癌细胞系SW48中转染ApoE过表达质粒及其特异性siRNA,实时荧光定量PCR（Real timeqPCR）检测转染ApoE的表达,利用Transwell小室观察ApoE对细胞侵袭迁移能力的影响,通过免疫印迹检测ApoE对基质金属蛋白酶（matrix metalloproteinase, MMP）家族蛋白表达的影响。结果 RTPCR检测结果显示：过表达ApoE组细胞中ApoE mRNA水平显著高于空白对照白组和阴性对照组,差异有统计学意义（P=0.008）,ApoE沉默组中ApoE mRNA水平与阴性对照组比,差异有统计学意义（P=0.013）;Transwell侵袭实验结果显示,穿膜细胞数：空白对照组、阴性对照组、过表达ApoE组、si-ApoE组依次为：（209±17）、（228±11）、（67±9）、（428±15）个/视野,过表达ApoE组和si-ApoE组与阴性对照组相比,差异有统计学意义（P<0.05）;Transwell迁移实验结果显示,空白对照组（326±18）个/视野,阴性对照组（338±21）个/视野,过表达ApoE组（133±11）个/视野,与阴性对照组相比,差异有统计学意义（P<0.05）,si-ApoE组（518±13）个/视野与阴性对照组相比,差异有统计学意义（P<0.05）。同时免疫印迹实验显示,过表达ApoE组细胞中MMP-2及MMP-9的表达下降,组织基质金属蛋白酶抑制剂2（tissue inhibitors of metalloproteinase-2, TIMP-2）的表达升高,而si-ApoE组MMP-2及MMP-9表达升高,TIMP-2的表达降低。结论 ApoE可影响结直肠癌细胞的侵袭、迁移能力,并且可能通过影响TIMP-2、MMP-2及MMP-9的表达发挥上述功能。     

Functionalization of liposomes with ApoE-derived peptides at different density affects cellular uptake and drug transport across a blood-brain barrier model

A promising strategy to enhance blood-brain barrier penetration by drugs is the functionalization of nanocarriers with uptake-facilitating ligands. We studied the cellular uptake, by cultured RBE4 brain capillary endothelial cells, of nanoliposomes (NLs) covalently coupled with monomer or tandem dimer of apolipoprotein E (ApoE)-derived peptides (residues 141-150), at various densities. NLs without functionalization did not show either relevant membrane accumulation or cellular uptake, as monitored by confocal microscopy and quantified by fluorescence-activated cell sorting. Functionalization with peptides mediated an efficient NLs uptake that increased with peptide density; NLs carrying monomeric peptide performed the best. Moreover, we studied the ability of ApoE-NLs to enhance the transport of a drug payload through a RBE4 cell monolayer. The permeability of a tritiated curcumin derivative was enhanced after its entrapment into ApoE-NLs, in particular those functionalized with the dimer (+83% with respect to free drug, P < 0.01). Thus, these NLs appear particularly suitable for implementing further strategies for drug brain targeting.

From the Clinical Editor

Re and her collaborators present a method for delivering nanoliposomes via the blood brain barrier by utilizing peptide fragments including monomers or tandem dimers ApoE. This method may enable enhanced nanoliposome associated drug delivery via the blood-brain barrier, which would have enormous significance in neurodegenerative and other CNS disorders.     

ApoE基因多态性与长寿及长寿相关表型的关联研究及Meta分析

目的 该研究在多群体中筛查验证ApoE基因多态性与长寿的关联,并调查ApoE基因变异与长寿相关表型的关联.方法 该研究采用群体1筛查,群体2、3验证的策略,共收集814个长寿老人(年龄≥90岁)和1136个无长寿家族史、当地一般人群的对照组(年龄30～65岁).通过PCR-RFLP和测序方法对ApoE基因进行基因分型.结果 在3个群体中长寿老人的ApoE ε3/ε3频率明显高于对照人群,与长寿存在正关联,这个结果与以往其他群体的研究结果并不相同;而长寿老人中ApoE ε3/ε4 频率明显低于对照群体,与长寿存在负关联.同时,在长寿老人中,ApoE ε3/ε3携带者比未携带ApoEε3/ε3者的高密度脂蛋白水平高(F=6.970,P=0.005),ApoE ε4携带者(ε3/ε4和ε4/ε4基因型总和)比非ApoEε4携带者的总胆固醇和低密度脂蛋白水平高(F=4.810,P=0.003和F=4.21 8,P=0.012).另外,该文对世界不同群体进行Meta分析,结果显示ApoE ε4携带者与长寿负关联(合并OR=0.42,95%CI:0.36～0.49),是长寿的风险因素.ε3/ε3基因型与长寿正关联(合并 OR=1.47,95%CI:1.25～1.74),是长寿的保护因素.结论 多个群体验证ApoE ε3/ε3可能是长寿的保护因素,这与以往报道ApoE ε2/ε2是长寿保护因素的结果不同;而ApoEε4是长寿的风险因素.

Abstract：

Objective The present study is to identify and replicate the association of ApoE longevity in multi-populations, and observe the association of ApoE with longevity-related phenotype. Methods By multi-population design:identifying ApoE gene associated with longevity in population 1, and replicating ApoE gene associated with longevity in population 2 and 3. A total of 814 "longevity cases" were classified as participants who had survived to age 90 years or more, with a total of 1136" younger controls" less than 65 years of age. ApoE gene was genotyped by PCR-RFLP and sequencing. Results Results showed the homozygous ε3/ε3 genotype in long-lived population was significantly higher frequent than those in controls,positively associated with longevity, whereas the heterozygous ε3/ε4 genotype were significantly lower frequent than those in controls, negatively associated with longevity in 3 different populations. ApoE ε3/ε3 was associated with higher HDL-C levels(F=6.970,P=0.005),and ApoE ε4-carrier(the ε3/ε4 and ε4/ε4 genotypes) was associated with significantly higher TC and LDL-C levels(F=4.810,P =0.003 and F=4.218,P=0.012)in long-lived individuals.In addition, the meta-analysis in 14 populations in world suggested ε4-carriers were negatively associated with longevity(pool ORs=0. 42,95% CI:0. 36～0.49);whereas the ApoE ε3/ε3 genotype was positively associated (pool ORs=1.47,95% CI:1.25~1.74) with longevity. Conclusions It was confirmed ApoE gene was associated with longevity in multi-population. ApoE ε3 gene could conferred protective effect for healthy longevity which was inconsistent the previous results published , whereas ApoE ε4 gene increased the risk effect for successful aging and longevity.     

中国人载脂蛋白E基因多态性与脑出血关系的Meta分析

目的 探讨中国人群载脂蛋白E(ApoE)基因多态性与脑出血的相关性.方法 通过文献检索收集脑出血组和对照组的病例对照研究,剔除不符合要求的文献,以漏斗图检验人选文献的发表偏倚,并根据各入选文献结果的同质性检验结果进行数据合并,计算总OR值,Meta分析采用Review Manager 4.2版统计软件.结果 共有10篇符合条件的文献纳入分析,Meta分析结果表明,以野生型E3/3基因型为参照,携带E2/2,E2/3,E2/4,E3/4,E4/4基因型的个体发生脑出血的危险性的OR值和95%可信区间分别为1.25(0.61～2.58)、1.35(0.86～2.11)、1.89(1.08～3.29)、1.79(1.44～2.23)和1.97(0.91～4.28),携带E4的个体OR值和95%可信区间为1.01(1.40～2.22)(P＜0.01),携带E2的个体OR值和95%可信区间为1.45(0.99～2.14)(P＞0.05).结论 E4等位基因携带者可能是脑出血的遗传危险因素.     

GSK3: A possible link between beta amyloid peptide and tau protein

Tau is a neuronal microtubule-associated phosphoprotein that is highly phosphorylated by glycogen synthase kinase 3 (GSK3). Tau phosphorylation by GSK3 regulates tau binding to microtubules, tau degradation and tau aggregation. Tau phosphorylation is important in Alzheimer disease pathology and in other tauopathies. In Alzheimer disease, it has been proposed that the peptide beta amyloid promotes GSK3 activation, resulting in tau phosphorylation. In this work, we review the links between beta amyloid peptide, tau protein and GSK3 that occur in familial Alzheimer disease. We also discuss the possible links between GSK3 and sporadic Alzheimer disease. Finally, we include a brief review of the pathology of animal models overexpressing GSK3.     

蛋氨酸诱导ApoE基因敲除小鼠主动脉不稳定斑块形成模型的研究

目的通过2%（wt/vol）L-蛋氨酸负荷诱导ApoE基因缺陷小鼠,建立高同型半胱氨酸（Hcy）血症的动脉粥样硬化（AS）易损斑块模型。方法将60只6周龄C57BL/6J系ApoE基因缺陷小鼠随机分为三组,每组20只。对照组：给予同剂量饮用水灌胃;高蛋氨酸组：给予2%（wt/vol）L-蛋氨酸灌胃;极高蛋氨酸组：给予[3%（wt/vol）L-蛋氨酸灌胃。另设正常小鼠作为正常对照组（n=20）。1个月和2月后分别测定血清Hcy和血脂变化,并通过斑块HE、VanGieson（EVG）弹性纤维染色和EVG苦味酸复红法胶原染色及anti-MAC3单克隆抗体标记来检测斑块大小、纤维帽厚度、胶原比例和炎症浸润程度的变化。TUNEL法检测主动脉血管组织中细胞凋亡情况。结果高蛋氨酸负荷可诱导ApoE基因缺陷小鼠AS易损斑块产生,且随负荷提高和时间延长程度加重,与对照组比较差异有统计学意义（P〈0.05）;凋亡细胞在脂核区大量积聚;内皮细胞水肿变性;血管平滑肌细胞（SMCs）由收缩型向合成型转变,胞质疏松,水肿变性,内质网扩张,胞质内空泡增多以及肌丝溶解;并可见核固缩的凋亡SMCs。结论高蛋氨酸饮食可诱导apoE基因缺陷小鼠出现高Hcy血症,从而生成AS易损斑块。斑块内SMCs增生与凋亡现象并存。     

ApoE与SLCO1B1基因多态性与老年脑梗死患者颅内动脉粥样硬化狭窄的相关性

目的 分析ApoE与SLCO1B1基因在老年脑梗死患者颅内动脉粥样硬化狭窄中的分布情况,及动脉狭窄程度与血脂的关系.方法 选取2018年2月至2019年2月北京中医药大学东方医院脑动脉粥样硬化患者200例为脑梗死组,以北美症状性颈动脉内膜剥脱术研究法分级标准划分动脉粥样硬化狭窄分级,其中无狭窄患者26例、轻度狭窄患者4...     

脂欣康对ApoE基因敲除小鼠海马形态学及胆碱乙酰转移酶和突触素表达的影响

目的研究脂欣康对ApoE基因敲除（ApoEKO）小鼠海马形态学及脑胆碱乙酰转移酶（ChAT）和突触素（P38）表达的影响。方法以ApoEKO小鼠为研究对象,随机分为ApoEKO组、脂欣康组、联合干预组,分别给予生理盐水、脂欣康、脂欣康＋维生素E灌胃,每日一次,连续灌胃60d,以同龄C57BL/6J小鼠作为正常对照组,之后将所有小鼠断头处死,采用HE染色观察各组小鼠海马神经元组织形态学改变,采用免疫组化技术与计算机图像分析技术检测ApoEKO小鼠海马内ChAT和P38的表达。结果与C57BL/6J小鼠相比,ApoEKO组小鼠海马区神经细胞结构破坏,ChAT阳性神经细胞数目与P38阳性染色颗粒明显减少,染色变浅,ChAT与P38平均灰度值明显增高（P〈0.05）。干预组的反应结果介于两者之间,联合干预组表达优于脂欣康组。结论脂欣康能够减轻ApoEKO小鼠海马区神经损害,增强ChAT与P38的表达,与维生素E间有协同作用。