三磷酸腺苷结合盒转运体A1对糖尿病地鼠巨噬细胞内胆固醇外流的影响

目的探讨糖尿病地鼠巨噬细胞内胆固醇外流的改变,尤其是三磷酸腺苷结合盒转运体A1(ABCA1)的功能变化以及与载脂蛋白AI(apoA-I)的相互作用。方法用金黄地鼠高脂及糖尿病模型,分离纯化腹腔巨噬细胞,用外源性apoA-I及8-br-cAMP在体外干预巨噬细胞,测定干预前后巨噬细胞ABCA1表达和细胞内胆固醇含量的变化。结果细胞内胆固醇含量在糖尿病组高于高脂组和正常对照组。apoA-I和8-br-cAMP在体外均增加糖尿病组巨噬细胞ABCA1 mRNA的表达。apoA-I使各组细胞内胆固醇外流均增加,且与孵育时间成正比,胆固醇外流量糖尿病组高于高脂组和正常对照组。结论糖尿病鼠巨噬细胞内胆固醇大量沉积,可能与内源性apoA-I量和功能的改变所引起的ABCA1介导的细胞内胆固醇外流受阻有关。     

2型糖尿病患者的高密度脂蛋白和乏脂血清对细胞内胆固醇外流的影响

目的 研究2型糖尿病患者高密度脂蛋白（HDL）及乏脂血清（LPDS）介导的细胞内胆固醇外流变化。方法 选取2型糖尿病患者13例和正常对照组17例,分离HDL和LPDS。利用人皮肤纤维母细胞（HSF）和人肝癌细胞（HepG2细胞）为实验细胞系,测定ATP结合盒受体A1（ABCA1）、ATP结合盒受体G1（ABCG1）和清道夫受体B1（SR-B1）的mRNA和蛋白表达。以HDL、LPDS为接受体测定3H胆固醇在上述细胞的胆固醇流出率。 结果HepG2 细胞高表达SR-B1;HSF经22-羟基胆固醇刺激后高表达ABCA1。2型糖尿病患者的HDL诱导HepG2细胞的胆固醇流出率显著低于对照组（P＜0.001）;而LPDS介导的HSF细胞的胆固醇外流率在两组间无显著差异。 结论 2型糖尿病患者的HDL诱导HepG2细胞的胆固醇外流功能明显降低,提示HDL功能异常在糖尿病患者的细胞内胆固醇积聚及动脉粥样硬化发展过程中发挥重要作用。     

冠心病患者巨噬细胞肝X受体的活化与胆固醇外流

目的:研究冠心病患者巨噬细胞肝X受体（LXR）及其下游的一些目的基因表达和胆固醇外流特点。 方法:分离冠心病患者组和对照组外周血的单核细胞，并转化为巨噬细胞。在TO-90131７的刺激下，观察巨噬细胞的aopA-I介导胆固醇外流的变化和LXR以及其下游一些目的基因的mRNA表达。 结果:冠心病患者的巨噬细胞，其一些影响胆固醇代谢及炎症反应的基因表达受到了影响，aopA-I介导的胆固醇外流能力下调，对刺激LXR后的反应性也降低。 结论:冠心病患者的巨噬细胞胆固醇外流功能下降，其一些影响胆固醇代谢及炎症反应的基因表达发生了改变，对刺激LXR信号的反应性下降。提示巨噬细胞的LXR信号途径可能是冠心病发病机制中的重要环节，也是冠心病治疗潜在靶点。     

阿托伐他汀对巨噬细胞肝X受体表达及胆固醇外流的影响

目的:他汀类药物是目前治疗脂代谢的有效药物,通过研究阿托伐他汀对巨噬细胞的肝X受体(LXR)及其下游的一些目的基因的表达和胆固醇外流的影响,探讨他汀类药物对LXR信号系统的作用。方法:分离人外周血的单核细胞,并转化为巨噬细胞。在阿托伐他汀的作用下,观察巨噬细胞的aopA-I介导的胆固醇外流的变化和LXR以及其下游目的基因ABCA1、SREBP2、CETP、PLTP、apoE、MMP-9和MIP-1α的mRNA及蛋白LXRα、ABCA1、MMP-9和MIP-1α的表达。结果:阿托伐他汀上调LXRα、ABCA1、SREBP2、CETP、PLTP基因的表达,而下调MMP-9和MIP-1α基因的表达,同时增强aopA-I介导的胆固醇外流。结论:巨噬细胞在阿托伐他汀的作用下,胆固醇外流增强,这种效应与阿托伐他汀上调LXR及其下游的影响胆固醇代谢的目的基因有关,同时,也抑制一些炎症反应的基因的表达。提示他汀类药物的治疗作用,与其影响巨噬细胞LXR信号途径有关,从而影响泡沫细胞的形成。     

Ⅰ型子宫内膜样癌中ABCA1、ApoA1及SR-B1的表达及意义

目的探讨HDL脂质转运蛋白ATP结合盒转运体A1(ATP-binding cassette transporter A1, ABCA1)、载脂蛋白A1(apolipoprotein A1, ApoA1)和吸收蛋白B类1型清道夫受体(scavenger receptor class B type 1, SR-B1)是否在Ⅰ型子宫内膜样癌(endometrial carcinoma, EC)中存在表达失衡。方法选取80例Ⅰ型EC和47例对照增生改变子宫内膜组织,采用免疫组化EnVision两步法检测ABCA1和ApoA1的表达;分析Ⅰ型EC中ABCA1、ApoA1与SR-B1、ER、PR蛋白表达的相关性及与临床病理特征的关系。结果 (1)Ⅰ型EC中ABCA1阳性率为86.1%(62/72),低于对照增生改变子宫内膜组织(95.5%,42/44),差异有统计学意义(P0.05);Ⅰ型EC组织中ApoA1阳性率为63.8%(44/69),高于对照增生改变子宫内膜组织(53.3%,23/43),差异无统计学意义(P0.05);(2)在Ⅰ型EC中,ABCA1与SR-B1、ApoA1、ER之间呈负相关;ApoA1与PR之间呈正相关;(3)Ⅰ型EC中ABCA1和ApoA1蛋白表达与临床病理特征无明显相关性(P0.05)。结论与对照增生改变子宫内膜组织相比,Ⅰ型EC中可能存在HDL转运蛋白ABCA1和HDL吸收蛋白SR-B1的表达失衡。     

烟酸经LXRα/NPC1途径促进巨噬细胞溶酶体胆固醇外流

目的:探讨烟酸(NA)对巨噬细胞溶酶体游离胆固醇外流的作用及其机制。方法:以佛波酯诱导人单核-巨噬细胞THP-1形成的巨噬细胞为细胞模型,利用激光扫描共聚焦成像技术观察NA对经氧化型低密度脂蛋白(oxLDL)孵育的巨噬细胞溶酶体胆固醇外流的作用,以及烟酸腺嘌呤二核苷酸磷酸(NAADP)拮抗剂Ned-19、Ca²⁺螯合剂BAPTA、肝X受体α(LXRα)siRNA和尼曼-皮克C1蛋白(NPC1)siRNA对NA效应的影响。RT-qPCR和Western blot检测NA、Ned-19和BAPTA对LXRαmRNA和NPC1蛋白表达的影响。结果:NA呈剂量依赖性地促进oxLDL孵育的巨噬细胞溶酶体胆固醇外流;这种效应可被Ned-19和BAPTA显著抑制。NA可明显促进NPC1蛋白及LXRαmRNA表达;这种效应也可被Ned-19和BAPTA明显抑制。LXRαsiRNA可明显抑制NA对NPC1蛋白表达的促进作用。siRNA沉默LXRα和NPC1表达也可明显减弱NA促溶酶体胆固醇外流的效应。结论:NA可显著促进巨噬细胞溶酶体胆固醇外流,其机制可能与其增加NAADP的生成,...     

LXR-ABCA1/ABCG1通路对BCG感染的巨噬细胞凋亡和炎性反应的调控作用

目的 揭示LXR-ABCA1/ABCG1通路对BCG感染后巨噬细胞凋亡和炎性反应的调控作用。方法 采用T0901317预处理RAW264.7巨噬细胞2 h和BCG感染24 h,设置4个实验组：对照组、T0901317组、BCG感染组和T0901317+BCG感染组。采用Western blot方法检测凋亡相关蛋白cleaved-Caspase3、cleaved-Caspase8、cleaved-Caspase9和TLR信号通路相关蛋白TLR2、TLR4、MyD88、TRAF6、p-NF-κB p65、IRF3、TBK1的表达,采用ELISA方法检测细胞培养上清中促炎细胞因子TNF-α、IL-6、IL-1β的含量。结果 与对照组比较,BCG感染组凋亡相关蛋白cleaved-Caspase3、cleaved-Caspase8、cleaved-Caspase9及依赖MyD88途径蛋白TLR2、TLR4、MyD88、TRAF6、p-NF-κBp65的表达均上调（P<0.01）,促炎细胞因子TNF-α、IL-6、IL-1β表达水平上升（P<0.01）;与BCG感染组相比,T09013...     

肺炎衣原体下调THP-1源性巨噬细胞ABCA1、ABCG1表达的机制研究

目的:观察c-Jun氨基末端激酶(JNK)对肺炎衣原体(Cpn)诱导的THP-1源性巨噬细胞ATP结合盒转运蛋白A1(ABCA1)、ATP结合盒转运蛋白G1(ABCG1)和过氧化物酶体增殖物激活受体γ(PPARγ)表达的调控作用,探讨Cpn下调THP-1源性巨噬细胞ABCA1/ABCG1表达的信号转导机制。方法:将THP-1单核细胞诱导分化为巨噬细胞后,观察Cpn感染对细胞内ABCA1/ABCG1及PPARγ mRNA和蛋白表达的影响。并用不同浓度的JNK特异性抑制剂SP600125对细胞进行预处理,观察SP600125对Cpn诱导的ABCA1/ABCG1、PPARγ mRNA和蛋白表达的影响。分别用RT-PCR和Western blotting检测各组ABCA1/ABCG1及PPARγ mRNA和蛋白表达。结果:Cpn下调THP-1源性巨噬细胞ABCA1/ABCG1及其上游调控基因PPARγ mRNA和蛋白表达。而SP600125能呈浓度依赖性地抑制Cpn感染所带来的上述影响。结论:Cpn可能通过JNK-PPARγ信号转导通路下调AB-CA1/ABCG1表达,减少巨噬细胞内胆固醇流出,促进动脉粥样硬化发生发展。     

ATP结合盒转运子A1与胆固醇外流

高密度脂蛋白(HDL)及其载脂蛋白具有抗动脉粥样硬化作用;这种血管保护作用主要是通过胆固醇逆转运来实现的.胆固醇外流是胆固醇逆转运的第一步,目前已明确ATP结合盒转运子A1(ABCA1)在胆固醇外流方面起关键作用.有关ABCA1的作用机制、表达调控和相互作用等方面的研究,近年来取得了很大的进展.对ABCA1的深入研究有利于进一步阐明脂质代谢过程,为防治脂质异常血症和动脉粥样硬化提供新思路和新手段.     

ABCG2/Bcrp1可能是大鼠气管上皮干细胞的标记物

目的： 探索气管上皮干细胞的表面标记物。方法: 取2周龄的Wistar大鼠，取出气管环， 5-FU损伤12 h后，取气管上皮涂片进行增殖细胞核抗原、ABCG2/Bcrp1抗原染色、RT-PCR检测ABCG2/Bcrp1基因表达。结果: 5－FU损伤后大鼠气管上皮气管环的上皮全部脱落，残留间隔分布的裸核样细胞，基底膜完整。气管上皮涂片免疫组织化学染色可见PCNA染色阳性的细胞与阴性细胞间隔分布，间接免疫荧光染色可见ABCG2/Bcrp1阳性细胞；RT-PCR检测ABCG2/Bcrp1阳性反应产物与预计长度符合。结论: ABCG2/Bcrp1可以作为气管上皮干细胞的标记，为进一步分离纯化扩增奠定基础。     

用核酸序列测定1型糖尿病HLA-DPB1和DQB1第2外显子基因

目的 研究山东省汉族1型糖尿病与HLA－DPB1和HLA－DQB1等位基因的相关性。方法 采用基于核酸序列测定的基因分型技术对52例1型糖尿病患者及38例正常对照进行了DPB1和DQB1基因分析。结果 DPB1＊2201（P＜0．01）和DQB1＊0201（P＜0．01）、＊0303（P＜0．05）及＊0604（P＜0．05）等位基因频率在糖尿病患者组显著高于对照组,而PB1＊0402（P＜0．01）和DQB1＊0301（P＜0．01）等位基因在糖尿病患者组显著低于对照组。结论 DPB1＊2201和DQB1＊0201、＊0303及＊0604等位基因可能是山东省汉族1型糖尿病的易感性等位基因,而DPB1＊0402和DQB1＊0301等位基因可能是山东省汉族1型糖尿病的保护性等位基因。     

Crystallization of oral fluid components in patients with type 1 diabetes mellitus

We systematized and described morphological criteria characterizing crystalline aggregates in mixed saliva from patients with type 1 diabetes mellitus.__________This revised version was published online in August 2005 with the addition of the issue titleTranslated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 139, No. 1, pp. 22–24, January, 2005     

Paraoxonase-1 is not associated with coronary artery calcification in type 2 diabetes: Results from the PREDICT study

Objectives: To determine any association between serum paraoxonase-1 (PON1) activity, protein and coding region genetic polymorphisms and coronary artery calcification (CACS) and to determine factors which modulate serum PON1 in type 2 diabetes (T2DM). Methods and results: 589 patients (419 Caucasian, 120 South Asian, 50 other) from the PREDICT Study were investigated. All patients were asymptomatic for coronary disease and had established T2DM. CACS, lipids, lipoproteins, inflammatory markers, insulin resistance and PON1 activity, concentration and Q192R and L55M genotypes were measured. Independent associations were: 1) PON1 activity negatively with insulin resistance, triglycerides and PON1-55 genotype and positively with PON1-192 genotype; 2) PON1 concentration negatively with Caucasian ethnicity, duration of diabetes and statin use and positively with plasma creatinine and PON1-192 genotype. There was no association between CACS and any of the PON1 activity, concentration or genotype and this finding was not different in the various ethnic groups within the PREDICT study. Conclusion: PON1 is modulated by a number of factors, some of which are reported here for the first time, including ethnicity and insulin resistance in subjects with T2DM. No association between CACS and PON1 was found.     

DC-SIGN polymorphisms are associated to type 1 diabetes mellitus

Type I diabetes mellitus (T1DM) is an autoimmune disorder featured by raised glucoses levels. It has been hypothesised that raised glucose levels in T1DM might be recognised as PAMPs, leading to immune response by overloading the cell receptors for pathogens recognition. DC-SIGN is a transmembrane protein, present in dendritic cells (DC) and macrophages: it has an important role in inflammatory response and T cells activation. Notably, DC-SIGN activation and triggering of the immune response depend on the type of ligand, which may lead to a pro or anti-inflammatory pathway. In our association study, we analysed the SNPs rs4804803 (−336 A>G) and rs735239 (−871 A>G), both at DC-SIGN promoter region, in 210 T1DM patients and 157 healthy controls, also looking for a correlation with the age of onset of the disease. We found that the allele G and genotypes G/G and A/G of SNP-871 (rs735239), as well as the alleles G-G (rs735239-rs4804803) and genotypes combined AA-GG (rs735239-rs4804803) were associated with protection of T1DM development. We did not find association between these variations with the age of onset of the disease and the presence of other autoimmune disorders. Our results suggest that SNPs in DC-SIGN promoter region can be associated to protection for T1DM in the Northeast Brazilian population.     

Good glycaemic control is associated with a better prognosis in breast cancer patients with type 2 diabetes mellitus

Although diabetes mellitus (DM) is one of the risk factors associated with increased breast cancer (BC) mortality, the effects of glycaemic control on the prognosis of BC have not been thoroughly evaluated. This retrospective study aimed to evaluate the relationship between glycaemic control and BC prognosis and to determine an optimal target of glycaemic control for BC patients with diabetes. We included 2812 stage 0–3 BC women, of whom 145 were diabetic and were 2667 non-diabetic. In those with diabetes, a mean haemoglobin A1C (HbA1C) <?7% (n?=?77) was defined as well-controlled diabetes, while a mean HbA1C >?9% (n?=?16) was defined as poorly controlled diabetes. All of the BC populations were followed from the date on which BC was diagnosed until 31 December 2015. Cox regression analysis was performed to estimate the adjusted hazards for all-cause mortality and BC-specific mortality. After controlling for the baseline and BC-related confounders, the adjusted hazard ratio (HR) for all-cause mortality and the HR for BC-specific mortality were 3.65 (95% confidence interval [95% CI] 1.13–11.82) and 8.37 (95% CI 1.90–36.91), respectively, for poorly controlled diabetic women and non-DM women. However, for the diabetic women with good glycaemic control, the HRs of all-cause mortality and BC-specific mortality were not significantly different (HR 0.91, 95% CI 0.42–1.01; HR 0.77, 95% CI 0.18–3.32, respectively) from those for both mortalities in non-DM patients. For moderate controlled diabetic women, the HRs for all-cause mortality and BC-specific mortality were 1.95 (95% CI 0.89–4.27) and 3.55 (95% CI 1.369–9.30), respectively. This pilot and retrospective cohort study reveals a relationship between glycaemic control and BC prognosis in diabetic women. In addition, well-controlled HbA1C, with maintained mean HbA1C values under 7%, may be associated with a better progression outcome of BC.     

Impairment of the ABCA1 and SR-BI-mediated cholesterol efflux pathways and HDL anti-inflammatory activity in Alzheimer's disease

The aim of our study was to investigate the effect of Alzheimer's disease (AD) on the cholesterol efflux capacity and anti-inflammatory activity of HDL. HDL and apoA-I were isolated from 20 healthy subjects and from 39 AD patients. Our results showed that serum- and HDL-mediated cholesterol efflux is significantly impaired in AD patients. This impairment of serum and HDL cholesterol efflux capacity was significantly inversely correlated to the AD severity as evaluated by MMSE scores. Results obtained from SR-BI-enriched Fu5AH and ABCA1-enriched J774 cells revealed that AD impaired the interaction of HDL and apoA-I with both the ABCA1 transporter and SR-BI receptor. Purified apoA-I from AD patients also failed to remove free excess cholesterol from ABCA1-enriched J774 macrophages. Interestingly, the decrease in plasma α-tocopherol content and the increase in MDA formation and HDL relative electrophoretic mobility indicated that AD patients had higher levels of oxidative stress. The anti-inflammatory activity of HDL was also significantly lower in AD patients as measured by the level of ICAM-1 expression. In conclusion, our study provides evidence for the first time that the functionality of HDL is impaired in AD and that this alteration might be caused by AD-associated oxidative stress and inflammation.