The work was aimed to investigate the differential expressions of lipid metabolism related genes in the early stage of atherosclerosis in the young apolipoprotein E deficient (apoE(-/-)) mice at different ages with normal chow diet. The genotypes of mice were identified by using multiplex polymerase chain reaction (multi-PCR) analysis. The semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and real-time quantitative RT-PCR were used to analyze the expressions of lipid metabolism related genes in the liver of apoE(-/-) and age-matched wild type (WT) mice of 14-day old, 1-month old, 2-month old, 3-month old. The serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) contents were assayed using COD-PAP and GPO-PAP methods. The serum apolipoprotein B100 (apoB100) content was quantitated by immune turbidimetry. The hearts were perfusion-fixed in 4% formaldehyde, infiltrated with 30% gum sucrose for 24 h at 4 °C, and embedded in OCT compound. The aortic sinus tissues were serially sectioned at -15 °C, stained with Sudan IV, and counterstained with light green. The results were shown as follows. Compared with that in WT mice, the mRNA levels of apoA I and apoA IV in apoE(-/-) mice aged from 14-day old to 3-month old changed prominently (P<0.05), with apoA I up-regulated and apoA IV down-regulated. At the age of 1 month, the expression of apoB100 in apoE(-/-) mice was higher than that in WT mice (P<0.05). The expression of apoA V was up-regulated (P<0.05) and there was obvious lipid deposition in the aortic intima in apoE(-/-) mice at the age of 2 months. The expressions of fatty acid translocase (Fat/CD36) and angiopoietin-like protein 3 (Angptl 3) in apoE(-/-) mice were higher than those in WT mice at the age of 3 months (P<0.05), while the expressions of peroxisome proliferator-activated receptor α (PPARα), liver X receptor α (LXRα), carnitine palmitoyl transferase I (CPT I) and acyl coenzyme A oxidase 1 (ACOX1) showed no significant changes. The serum TC, TG, LDL-C and HDL-C contents in apoE(-/-) mice aged from 14-day old to 3-month old were higher than those in age-matched WT mice. apoE(-/-) mice showed a marked increase in serum apoB100 content, consistent with the trend of serum LDL-C content and apoB100 mRNA content in the liver. The results suggest that the mRNA expressions of apoA I, apoA IV, apoA V, apoB100 and Angptl 3 in apoE(-/-) mice change significantly compared with those in WT mice, and these genes might be relevant to the complicated lipid metabolism network, and involved in the early stage of atherogenesis.
Animals ; Apolipoprotein A-I ; metabolism ; Apolipoprotein B-100 ; blood ; Apolipoproteins A ; metabolism ; Apolipoproteins E ; genetics ; Atherosclerosis ; genetics ; Gene Expression ; Lipid Metabolism ; genetics ; Lipoproteins, HDL ; blood ; Lipoproteins, LDL ; blood ; Liver ; metabolism ; Mice ; Mice, Knockout ; Triglycerides ; blood

OBJECTIVETo identify potential mutation of apolipoprotein E (apoE) gene in a male patient affected with lipoprotein glomerulopathy (LPG), his mother and his sister.

METHODSThe patient and his mother both had histologically confirmed LPG. His sister and his father were asymptomatic. Genomic DNA was extracted from peripheral blood samples. PCR products of the coding region of exons 3 and 4 of the apoE gene were cloned into a pTA2 vector and sequenced. Genetic variations of the apoE gene were detected using PCR and restriction fragment length polymorphism (RFLP).

RESULTSAn apoE gene mutation was identified in the patient's family. Sequence analysis confirmed a 9-bp deletion in the exon 4 of apoE gene from nt 484 to 492. The 9-bp deletion resulted in loss of 3 amino acids at positions 143-145. The sister of the propositus carried the same mutation, though she had neither proteinuria nor elevated plasma apoE. Sequence analysis of exon 3 showed no abnormality. No abnormalities were found in the father's apoE gene sequence. Analysis of genetic variations of the apoE gene by PCR and RFLP confirmed a 57 bp fragment consistent with the 9-bp deletion in exon 4. The father had a normal ε 3 ε 3 genotype.

CONCLUSIONThe 9 bp deletion of apoE may be associated with the pathogenesis of LPG.

Adolescent ; Apolipoproteins E ; blood ; genetics ; Exons ; Female ; Genetic Predisposition to Disease ; Genetic Variation ; Humans ; Kidney Diseases ; blood ; genetics ; Kidney Glomerulus ; metabolism ; pathology ; Lipoproteins ; blood ; Male ; Mutation ; Pedigree

OBJECTIVES: Plasma lipid profiles and Apolipoprotein E (ApoE) are established risk factors for cardiovascular disease (CVD). The knowledge of lipid profile may estimate the potential victims of cardiovascular disease before its initiation and progression and offers the opportunity for primary prevention. The most common ApoE polymorphism has been found to influence plasma lipid concentrations and its correlation with CVD has been extensively investigated in the last decade. METHODS: The ApoE polymorphism and its influence on plasma lipid were investigated in healthy woman workers. The information on confounding factors was obtained through a self-administered questionnaire and ApoE polymorphism was investigated using PCR. RESULTS: The relative frequencies of alleles E2, E3 and E4 for the study population (n=305) were 0.127, 0.750 and 0.121, respectively. ApoE polymorphism was associated with variations in plasma HDL-cholesterol lipid profile. In order to estimate the independent effects of alleles E2 and E4, as compared with E3, on lipid profile, multiple regression was performed after adjustment for confounding variables such as age, BMI, blood pressure, education status, insulin, fasting glucose, HOMA-IR, menopause. ApoE2 had a negative association with HDL cholesterol and ApoE4 had a positive association with LDL cholesterol. CONCLUSIONS: This study identified that the ApoE and CVD risk factors contribute to the lipid profiles, similar to other studies. The analysis including dietary intake and other gene in further studies may help to identify clear effects on lipid profiles as risk factor for CVD.
Adult ; Apolipoproteins E/blood/*genetics/metabolism ; Cardiovascular Diseases/epidemiology/prevention & control ; Cholesterol, HDL/genetics ; Female ; *Genotype ; Humans ; Lipid Metabolism/*genetics ; Polymerase Chain Reaction ; Primary Prevention

OBJECTIVECoronary heart disease (CHD) is one of the most common causes of death in the world. Some studies suggested that CHD begins in childhood. Obesity and dyslipidemia are important risk factors of coronary heart disease. Apolipoprotein (apo)E gene associated with dyslipidemia and coronary heart disease. The present study was designed to investigate the expression status of apoE gene in peripheral blood monocyte and association of apoE gene expression with lipids in children with obesity.

METHODSAmong 32 children with obesity and 32 healthy children without obesity or overweight, ApoE gene expressions were determined by competitive reverse transcription-polymerase chain reaction in peripheral blood monocyte. The concentrations of plasma triglyceride, total cholesterol, low density lipoprotein-cholesterol, high density lipoprotein-cholesterol, lipoprotein(a), apoA I, apoB(100) and apoE were measured.

RESULTSExpression of apoE gene was detected in peripheral blood monocyte. Expression of apoE gene was significantly reduced in children with obesity as compared with control group (0.29 +/- 0.14 moles/mole GAPDH mRNA vs. 0.36 +/- 0.10 moles/mole GAPDH mRNA, t = 2.15, P < 0.05). The more severe was the degree of obesity, the more significantly reduced the expression of apoE gene; the degree of obesity was negatively correlated with the levels of expression of apoE gene (correlation coefficient = -0.40, P < 0.05). Compared with control group, the levels of triglyceride, total cholesterol, low density lipoprotein-cholesterol, and apoB(100) were higher, and those of high density lipoprotein-cholesterol, apoA I and apoE were lower in children with obesity [(1.68 +/- 0.50) mmol/L vs. (0.99 +/- 0.54) mmol/L, (4.47 +/- 0.91) mmol/L vs. (3.33 +/- 0.90) mmol/L, (2.23 +/- 0.71) mmol/L vs. (1.13 +/- 0.96) mmol/L, (94.48 +/- 9.97) mg/dl vs. (83.81 +/- 15.64) mg/dl, (1.47 +/- 0.39) mmol/L vs. (1.73 +/- 0.36) mmol/L, (112.71 +/- 27.86) mg/dl vs. (134.80 +/- 45.36) mg/dl, (24.50 +/- 10.92) mg/L vs.(35.07 +/- 9.79) mg/L, respectively, P < 0.05]. ApoE gene expression was associated with plasma lipids metabolism in children with obesity. The quantity of apoE gene expression was inversely associated with low density lipoprotein-cholesterol, positively correlated with apoE (correlation coefficient = -0.33, 0.35, respectively, P < 0.05). The quantity of apoE gene expression was not associated with total cholesterol, triglyceride, high density lipoprotein-cholesterol, lipoprotein(a), apoA I, and apoB(100) (correlation coefficient = -0.19, -0.11, 0.16, 0.09, 0.18, 0.22, P > 0.05).

CONCLUSIONExpression of apoE gene was significantly reduced in peripheral blood monocyte in children with obesity. The quantity of apoE gene expression was associated with degree of obesity and abnormality of blood lipids.

Apolipoproteins E ; genetics ; Child ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Gene Expression ; genetics ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Obesity ; blood ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Triglycerides ; blood

OBJECTIVETo explore the influence of apolipoprotein E (ApoE) gene polymorphism on the lipid metabolism regulatory effect of Xuezhikang Capsule (XZKC).

METHODSApoE polymorphism of 74 patients with hyperlipidemia was detected by gene sequencing method, and their plasma levels of total cholesterol (TC), triglyceride (TG), high- and low-density lipoprotein cholesterol (HDL-C and LDL-C) were determined before and after they received a 6-week treatment of XZKC, for analyzing the relationship between ApoE gene polymorphism and the changes of various blood lipids associated indices.

RESULTSThe effect of XZKC on reducing TG in the epsilon2 allele (E2/E2 and E2/E3 genotypes) was higher than that in the E3/E3 genotypes and epsilon4 allele (E3/E4 and E4/E4 genotypes), while on increasing HDL-C, it showed more effect in the epsilon4 allele (E3/E4 and E4/E4 genotypes) than that in the epsilon2 allele (E2/E2 and E2/E3 genotypes) and E3/E3 genotypes.

CONCLUSIONPatients' ApoE gene polymorphism could influence the lipid regulatory effect of XZKC, embodying mainly by raising HDL-C and reducing TG in patients with different ApoE genotypes to different extents.

Adult ; Aged ; Apolipoproteins E ; genetics ; Cholesterol, HDL ; blood ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Genotype ; Humans ; Hyperlipidemias ; blood ; drug therapy ; genetics ; Lipid Metabolism ; drug effects ; Male ; Middle Aged ; Polymorphism, Genetic ; Triglycerides ; blood

BACKGROUNDLipoprotein glomerulopathy (LPG) is a renal disease characterized by thrombus-like lipoproteins in the glomerular capillaries and its abnormal lipoprotein profiles with marked elevation of apolipoprotein E (apoE). In this study, 15 Chinese patients with LPG were involed in exploring the association of the genetic variation and its plasma level in the pathogenesis of LPG.

METHODSA retrospective analysis of the clinical and pathological features was made in 15 patients with LPG. Plasma concentrations of apoE were measured with radial immunodiffusion assay. Genetic variations of apoE gene were detected using polymerase chain reaction and restriction fragment length polymorphism. Glomerular deposition of apoA, apoB and apoE in these patients were detected by immunofluorescence staining using monoclonal antibodies.

RESULTSBiochemical profiles of lipids and lipoproteins revealed markedly elevated levels of triglyceride, apoB and apoE, but approximately normal levels of total cholesterol, apoA1 and lipoprotein(a) [Lp(a)], which resembled familial hypertriglyceridemia. Genetic analysis demonstrated that the genotype distribution of apoE were 7 cases with epsilon3/epsilon4, 4 cases with epsilon3/epsilon3 and 2 cases with epsilon2/epsilon3. The other 2 cases (a mother and her son) showed a same distinct band. The band pattern of later 2 cases was quite similar to the apoE variant of Tokyo type. The calculated allele frequency of epsilon 4 was relatively high in cases with LPG in comparison with that in the normal controls. We further divided the 13 patients into three groups according to their genotypes of apoE. Patients with the genotype of apoE epsilon2/epsilon3 showed a lower level of plasma apoE as compared to those with apoE epsilon3/epsilon4 (P < 0.05). The serum level of high-density lipoprotein (HDL) was the lowest in patients with the genotype of apoE epsilon3/epsilon4. No difference was found among the patients with different apoE genotype in the other clinical and pathological characteristics.

CONCLUSIONSThe genotype of apoE epsilon3/epsilon4 is the predominant one in Chinese patients with LPG. Patients with this genotype tend to have a higher plasma level of apoE and more severe lipid dysmetabolism. No correlation was found between the genotype of apoE and the clinical features in patients with LPG.

Adolescent ; Adult ; Apolipoprotein E2 ; Apolipoprotein E3 ; Apolipoproteins E ; blood ; genetics ; Child ; Female ; Genetic Variation ; Genotype ; Humans ; Kidney Diseases ; blood ; genetics ; pathology ; Kidney Glomerulus ; blood supply ; pathology ; Lipoproteins ; metabolism ; Male ; Middle Aged

OBJECTIVE: To explore the effect of chronic iron overload on the lesion of atherosclerosis (AS) in apolipoprotein (apo) E knockout mice. METHODS: Twenty-four ApoE knockout mice were randomly divided into ApoE knockout group (0.1 mL saline for 4 weeks) and iron overload group (10 mg iron dextran for 4 weeks). The levels of serum iron (SI), total iron binding capacity, contents of malondialdehyde (MDA), and activity of superoxide dismutase (SOD) in the liver were measured. Iron deposition in the liver and heart was determined, and atherosclerotic plaque areas of the sinus aortae were analyzed. RESULTS: In the iron overload group, the levels of SI increased by 377.86%, the saturation of transferrin increased by 121.98% and the levels of iron in the liver increased by 2,548.15% (P<0.01). The contents of MDA in the liver increased by 32.51% (P<0.01), and the activity of SOD in the liver decreased by 17.2% in the ApoE knockout group (P<0.05). The level of MDA in the liver increased by 411.15%, and the activity of SOD in the liver decreased by 46.84% in the iron overload group (P<0.01). There was a significant deposition of iron in the liver and heart of mice, and the areas of atherosclerotic plaque of sinus aortae increased markedly in the iron overload group. CONCLUSION Chronic iron overload may promote the development of AS lesion in the ApoE knockout mice, in which the increased oxidative stress and lipid oxidation may involve.
Animals ; Apolipoproteins E ; genetics ; metabolism ; Atherosclerosis ; etiology ; metabolism ; pathology ; Iron ; blood ; Iron Overload ; complications ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; metabolism ; Oxidative Stress ; Random Allocation ; Superoxide Dismutase ; metabolism ; Transferrin ; metabolism

OBJECTIVETo observe the regulatory effect of assorted use of Chinese drugs for detoxifying and activating blood circulation on serum high sensitive C-reactive protein (hs-CRP) in apolipoprotein E gene knock-out [ApoE (-/-)] mice.

METHODSApoE (-/-) mice of 13-week old were devided into two groups, 12 in Group A fed with common forage and 98 in Group B with high lipid forage. Besides, 12 C57 BL/6J mice, 13-week old, were set as Group C fed with common forage. After being fed for 19 weeks, and the formation of vulnerable plaque had been confirmed in 2 mice of Group B, the other 96 mice were sub-divided into 8 groups, 12 in each group. Except that 0.4 mL normal saline was infused to the Group B1 as well as Group A and C, the other 7 sub-groups of Group B were respectively medicated with various drugs as follows: B2, Polydatin (PD, an extract from giant knotweed rhizome for detoxicating) 26. 6 mg/kg; B3, Xiongshao Capsule (XS, a Chinese herbal preparation for activating blood circulation), 110 mg/kg; B4, PD 53.2 mg/kg + XS 220 mg/kg; B5, PD 26. 6 mg/kg + XS 110 mg/kg; B6, PD 13.3 mg/kg + XS 55 mg/kg; B7, Lovastatin 3.3 mg/kg; and B8, Xuezhikang (XZK, a Chinese patent drug) 0.2 g/kg, all were administered by dissolving in 0.4 mL of distilled water for gastrogavage. The serum contents of hs-CRP were detected, with blood sample drawing from inferior vena cava, using enzyme-linked immunosorbent assay 17 weeks after medication.

RESULTSThe hs-CRP level was significantly higher in Group B1 than in Groups A and C (P <0.05, P <0.01). Comparisons between various sub-groups of Group B in hs-CRP content showed that hs-CRP in Group B2, B4 and B7 was lower than that in B1 (P <0.01), hs-CRP in Group B4 was the lowest, and hs-CRP was lower in Group B2 than in Group B3.

CONCLUSIONAssorted use of Chinese drugs for detoxifying and activating blood circulation could reduce serum hs-CRP level in ApoE (-/-) mice.

Animals ; Apolipoproteins E ; genetics ; metabolism ; Atherosclerosis ; drug therapy ; genetics ; metabolism ; physiopathology ; Blood Circulation ; drug effects ; C-Reactive Protein ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Random Allocation

OBJECTIVETo investigate the effect of nano-TiO(2) intratracheal instillation on the progression of dyslipidemia and atherosclerosis in apolipoprotein E-knockout mice.

METHODSThe nano-TiO(2) was ultrasound with phosphate-buffered saline solutions (PBS) into its suspension for exposure. A total of 46 specific pathogen free (SPF) level of 11-week-old male apolipoprotein E-knockout mice were randomly divided into groups by their body weights: non-treatment group (8 mice), PBS control group (9 mice), high dose group (1.0 mg/ml, 10 mice), medium dose group (0.5 mg/ml, 10 mice), and low dose group (0.1 mg/ml, 9 mice). Except the non-treatment group, mice from other groups were intratracheally instilled with 0.05 ml each time, twice a week. After exposure of 6 weeks, viscera index, blood TC, TG, HDL-C, LDL-C, and organic lipid ratio were assessed as biomarkers. Artery and aortic root issues were assessed by histopathology.

RESULTSAfter 5 weeks exposure, mice body weights in high dose group ((29.7 ± 1.9) g) started to drop, compared to PBS control ((31.3 ± 1.9) g, t = -1.58, P < 0.05) and low dose group ((31.4 ± 1.4) g, t = -1.17, P < 0.05); after 6 weeks, high dose group ((28.8 ± 1.5) g) was lower than PBS control ((30.4 ± 1.9) g, t = -1.60, P < 0.05), non-treatment group ((30.2 ± 1.3) g, t = -1.43, P < 0.05) and low dose group ((30.6 ± 1.0) g, t = -1.83, P < 0.05). TC levels of non-treatment, PBS control, high dose group, medium dose group and low dose group were (2.92 ± 1.18), (3.12 ± 0.73), (4.19 ± 1.86), (3.46 ± 0.72) and (2.57 ± 0.64) mmol/L, respectively; TG levels were (0.39 ± 0.13), (0.39 ± 0.08), (0.60 ± 0.21), (0.55 ± 0.19) and (0.41 ± 0.11) mmol/L, respectively; HDL-C levels were (1.67 ± 0.45), (1.54 ± 0.67), (0.93 ± 0.50), (1.02 ± 0.48) and (1.31 ± 0.64) mmol/L; TG levels of high dose group were higher than that of non-treatment group (t = 1.27, P = 0.03) and low dose group (t = 1.62, P = 0.01); TG levels of medium dose group was higher than PBS control (t = 0.16, P = 0.04), and TC levels of high dose group were higher than PBS control (t = 0.22, P = 0.01), non-treatment group (t = 0.22, P = 0.04) and low dose group (t = 0.20, P = 0.03), and HDL-C levels of high dose group were lower than PBS control (t = -0.61, P = 0.04) and non-treatment group (t = -0.74, P = 0.04); organic lipid ratio of each group were (2.27 ± 0.51)%, (2.06 ± 0.53)%, (2.90 ± 0.50)%, (2.60 ± 0.23)%, (2.24 ± 0.45)%; high dose group were higher than PBS control (t = 0.85, P = 0.00), non-treatment group (t = 0.64, P = 0.03) and low dose group (t = 0.67, P = 0.01); medium dose group was higher than PBS control (t = 0.54, P = 0.02). The plaque lipid content and calcium content which showed the progression of atherosclerosis and plaque rupture were elevated in medium and high dose groups.

CONCLUSIONIntratracheal instillation of nano-TiO(2) can induce dyslipidemia and accelerate the development of atherosclerosis and plaque rupture in ApoE-/-mice.

Animals ; Apolipoproteins E ; genetics ; Atherosclerosis ; blood ; chemically induced ; Dyslipidemias ; blood ; chemically induced ; Instillation, Drug ; Lipid Metabolism ; Lipids ; blood ; Male ; Mice ; Mice, Knockout ; Nanoparticles ; Specific Pathogen-Free Organisms ; Titanium ; administration & dosage ; pharmacology

OBJECTIVETo study the gene expression profile of liver of young apoE(-/-)/LDLR(-/-)/Lepr(db/db) treble genes mutant mice and disclose its relationship to hyperlipidemia and the following atherosclerotic lesion.

METHODSThe gene expression profile was investigated using cDNA microarray technique; the plasma total cholesterol(TC) and triglyceride(TG) levels were analyzed by COD-PAP and GPO-PAP method. And morphological observations of the aorta were made.

RESULTSAmong the 4000 target genes, 92 genes were up-regulated and 105 genes were down-regulated in the treble genes mutants, compared with wild type control. Among the differentially expressed lipid metabolism related genes, cholesterol synthesis gene coding for farnesyl diphosphate farnesyl transferase was down-regulated, while triglyceride metabolism gene e.g. pancreatic lipase related protein 1 gene (Pnliprp1) was up-regulated. Expression profile of carbohydrate, cell skeleton and immune related genes were also altered. On the other hand, in the plasma from the treble genes mutant mice at 5 weeks of age, hyperlipidemia was found to be combined with atheroslerotic lesion. All these biochemical and pathological changes were aggravated following aging.

CONCLUSIONThe data suggested that the multiple genes mutations, especially those involved in lipid metabolism, were contributing to the alteration of liver gene expression profile that might lead to hyperlipidemia and atherosclerotic lesion in the young apoE(-/-)/LDLR(-/-)/Lepr(db/db) mutants.

Animals ; Apolipoproteins E ; genetics ; Cholesterol ; blood ; Farnesyl-Diphosphate Farnesyltransferase ; genetics ; metabolism ; Female ; Gene Expression Profiling ; methods ; Hyperlipidemias ; blood ; genetics ; metabolism ; Lipase ; genetics ; metabolism ; Lipid Metabolism ; Male ; Mice ; Mice, Knockout ; Oligonucleotide Array Sequence Analysis ; methods ; Receptors, LDL ; genetics ; Receptors, Leptin ; genetics ; Triglycerides ; blood