OBJECTIVETo explore the relationship between microsomal triglyceride transfer protein (MTP) gene variation and diabetic dyslipidemia among Chinese.

METHODSUsing PCR restriction fragment length polymorphism (PCR-RFLP) analysis and gene sequencing, we studied the influence of a common MTP gene polymorphism in the promoter region on the apoB-containing lipoproteins in 44 Chinese type 2 diabetic subjects and 32 non-diabetic volunteers.

RESULTSA common functional G/T polymorphism in 493 bp upstream from the transcriptional start point was detected among native Chinese. There were 41 carriers (53.9%) of the MTP-493 G/G genotype, 28 (36.8%) of the MTP-493 G/T genotype and 7 (9.3%) of the MTP-493 T/T genotype. The allele frequency of MTP-493 T in the diabetic group was 0.30. The MTP-493 T/T diabetic group had significantly higher TG (P < 0.05), VLDL-CH (P < 0.05) and smaller LDL particle size (P < 0.001) than the MTP-493 common genotype group.

CONCLUSIONGenetic variation in the MTP promoter is likely to be highly involved in the production of dyslipidemia in type 2 diabetic subjects.

Carrier Proteins ; genetics ; Diabetes Mellitus, Type 2 ; genetics ; Genotype ; Humans ; Hyperlipidemias ; etiology ; genetics ; Polymorphism, Genetic ; Promoter Regions, Genetic

The aim of this paper was to investigate the mechanism of the active peptide DP17 of Eupolyphaga steleophaga in the treatment of hyperlipidemia rats. HPLC and MADIL-TOF/TOF-MS were used for the amino acid sequence analysis and solid-phase synthesis on the active peptide of E. steleophaga which were obtained by biomimetic enzymatic hydrolysis, separation and purification. The hyperlipidemia model was established by feeding with high-fat diet.Twenty days later, the rats in the blank group and the model group were given the saline and the rats in remaining groups were given the corresponding drugs by oral administration. After administration for 4 weeks, the levels of triglyceride(TG), total cholesterol(TC) and low density lipoprotein(LDL) in serum, the levels of TG, TC, adenosine monophosphate(AMP), adenosine triphosphate(ATP) in liver tissues and TG in feces were detected, respectively. Hematoxylin-eosin(HE) staining was used to observe the pathological changes of liver tissues. The Real-time fluorescence quantitative PCR method was used to detect the expression of acetyl coa carboxylase(ACC) and hydroxymethylglutaryl-coa reductase(HMGCR) mRNA in liver tissues. The expression of mammalian target of rapamycin(mTORC1) protein and adenosine 5'-monophosphate-activated protein kinase(AMPK) in liver tissues were detected by Western blot. The analysis showed that the amino acid sequence of active peptide from E. steleophaga was DAVPGAGPAGCHPGAGP(DP17). The results of pharmacological experiments showed that after oral administration of DP17 in rats, the levels of TG, TC and LDL in serum as well as TG and TC levels in liver tissues were significantly decreased(P<0.05), while the levels of AMP, ATP in liver tissues and TG content in feces were significantly increased(P<0.05); the liver steatosis of rats was significantly relieved; the expression of ACC, HMGCR mRNA and mTORC1 protein in liver tissues were significantly reduced, while the expression of AMPK phosphorylated protein was significantly increased(P<0.05). DP17, the active peptide of E. steleophag can significantly reduce lipid accumulation in liver tissues, and it may play a role in reducing blood lipids by regulating the energy metabolism balance in the body and activating AMPK/mTOR signaling pathway.
Animals ; Diet, High-Fat/adverse effects* ; Hyperlipidemias/genetics* ; Lipids ; Liver ; Peptides ; Rats ; Triglycerides

The aim of this paper was to study the improvement effect of ethanol extract from Citri Reticulatae Pericarpium(CRP) on triglyceride of hyperlipidemia model rats, and to explore the possible mechanism. SD rats were randomly divided into normal group, model group, positive control group, and high, medium and low-dose CRP ethanol extract groups, with 10 rats in each group. During the experiment, except for the normal group that was fed with distilled water and ordinary feed, rats in the other groups were given different concentrations of alcohol and fed with high-sugar and fat diets. All rats were given free diets. While being modeled, each group was administered with 0.01 mL·g~(-1) by gavage once a day for six weeks. Blood samples were collected after two weeks, four weeks and six weeks of drug treatment. After the completion of the experiment, blood, liver and adipose tissue were collected. Triglyceride(TG), alanine aminotransferase(ALT), aspartate aminotransferase(AST), alkaline phosphatase(ALP) in serum, TG in liver tissue and TG in fecal were detected. Free fatty acid(FFA) and triglyceride-related hydrolase, such as adipose tiglyceride lipase(ATGL), lipoprotein lipase(LPL), hepatic lipase(HL), hormone-sensitive triglyceride lipase(HSL) were detected by ELISA. The mRNA expressions of peroxisome proliferators-activated receptors(PPARγ), sterol regulatory element binding protein 1 c(SREBP-1 c) and farnesoid X receptor(FXR) were determined by RT-PCR. Compared with the model group, each administration group could reduce TG levels in serum and liver to varying degrees, reduce serum ALT, AST, ALP activities, significantly reduce free fatty acid content in serum, significantly increase triglyceride metabolism-related enzymes, including fat ATGL, LPL and liver HL content, and significantly reduced the content of fat HSL. According to the study of transcriptional regulation genes relating to triglyceride metabolism, extract from CRP could significantly increase the mRNA expressions of PPARγ and FXR. In conclusion, ethanol extract from CRP could ob-viously reduce the TG level of hyperlipidemia model rats, and might reduce plasma TG content by increasing PPARγ-LPL/ATGL and FXR-HL triglyceride hydrolysis pathways.
Animals ; Ethanol ; Hyperlipidemias/genetics* ; Liver ; Plant Extracts ; Rats ; Rats, Sprague-Dawley ; Triglycerides

Interleukin-8 is CXC chemokine that is initially discovered using chemotaxis and the activation of neutrophils and induces the migration and proliferation of smooth muscle cells. Interleukin-8 is a potent angiogenic factor that may play a role in atherosclerosis. To establish the temporal correlation between IL-8 expression and plaque development, we examined the expression during atherosclerosis of hyperlipemia rabbits using immunohistochemistry, ELISA, in situ hybridization. By location of immunohistochemistry, the expression of IL-8 protein increased obviously in intima of hyperlipemia rabbits at 8 and 12 week. Quantitative analysis of the expression of IL-8 Immunohistochemistry indicated that positive area of AS model was 4.48 times and 8.76 times that of control group at 8 and 12 week. The valuation of IOD of AS model was 4.16 times and 4.36 times that of control group at 8 and 12 week. By specific ELISA, the ratio of the IL-8 protein to total protein of AS model was 1.84 times and 2.06 times that of control group at 8 and 12 week. By location of in situ hybridization, positive location was strong in intima of hyperlipemia rabbits at 8 week. We observed the dynamic alteration of interleukin-8 protein and gene expression in atherosclerotic lesions of hyperlipemia rabbits with establishing model. Interleukin-8 protein and gene expression was up-regulation in the development of fatty streaks in hyperlipemia rabbit.
Animals ; Aorta ; metabolism ; Atherosclerosis ; etiology ; metabolism ; Hyperlipidemias ; complications ; Interleukin-8 ; biosynthesis ; genetics ; Male ; RNA, Messenger ; biosynthesis ; genetics ; Rabbits

OBJECTIVE: To construct a subtracted cDNA library of chronic intermittent hypoxia (CIH) rabbit liver by suppression subtractive hybridization (SSH). METHODS: Twenty-four rabbits were divided into 4 groups: ordinary feeding group, full-fat food group, ordinary feeding in chronic intermittent hypoxia group, and full-fat food in chronic intermittent hypoxia group. The mRNAs were extracted from different rabbit livers and converted into double-strand cDNA. After digestion with restriction enzyme, the cDNA of hyperlipidemia-sensitive rabbit group was subdivided into 2 portions and each one was lighted with different adaptors. Two rounds of both hybridization and suppression PCR obtained the differentially expressed cDNA. The PCR products were inserted into T/A vector to set up the subtractive cDNA library. The clones were selected and amplified by PCR and identified. RESULTS: Based on the pathology of the abdominal aorta and liver, and the amplified library contained 500 positive bacteria clones, including 462 clones, which had inserts from 250 to 700 bp by PCR analysis. A novel rabbit gene, Cthrc1, involved in CHI had been cloned. The GenBank Accession Number is XM_418373. CONCLUSION The molecular mechanism of CIH promoting atherogenesis formation is made clear.
Animals ; Gene Library ; Genetic Vectors ; Hyperlipidemias ; Hypoxia ; metabolism ; pathology ; Liver ; metabolism ; pathology ; Nucleic Acid Hybridization ; methods ; RNA, Messenger ; genetics ; Rabbits

Apolipoprotein E (apo E) plays a role in the regulation of the lipid metabolism of humans. Apo E, 229 amino acid polypeptide, is classified into three major isoform (E2, E3, E4) according to the differences of amino acid in position 112 and 158. In the normal population apo E3 isoform is most prevalent and apo E2 or E4 is frequently associated with hyperlipoproteinemia. To find out the frequency of apo E isoform distribution in the Korean population, apo E genotyping was performed. After amplification of apoE gene by polymerase chain reaction (PCR), restriction isotyping was done by cleavage with restriction enzyme Hha I and polyacrylamide gel electrophoresis. The apo E allele frequency in 73 normal subjects was 4.8% for E2, 84.9% for E3 and 10.3% for E4. In diabetic patient with hyperlipoproteinemia, the frequency of apo E allele was 6.3% for E2, 81.0% for E3 and 12.7% for E4. There was no significant difference in apo E isoform distribution between diabetics and normal populations. But in patients with cardiovascular disease with hyperlipidemia, the apo E4 allele frequency was significantly higher than normal (20.0% vs 10.3%, p<0.005). Apo E3 was the most common isoform in normal and diabetic subjects and apo E2 isoform was rather low frequency compared to Caucasians. This pattern is similar to the Japanese population but somewhat different from other populations. From the data of a high association of apo E4 allele and cardiovascular disease with hypercholesterolemia, apo E isoform may be one of the determinants of hyperlipoproteinemia. The PCR method may be useful in apo E genotyping.
Apolipoproteins E/*genetics ; Arteriosclerosis/*genetics ; Base Sequence ; Genotype ; Humans ; Hyperlipidemias/*genetics ; Korea/epidemiology ; Molecular Sequence Data ; Polymerase Chain Reaction ; Predictive Value of Tests ; Reference Values ; Restriction Mapping

OBJECTIVETo investigate the association between polymorphism of leukotriene A4 hydrolase (LTA4H) gene among ischemic stroke patients and the related subtypes in the discordant sib pairs.

METHODSFamilies including ischemic stroke patients and their siblings were recruited. Four single nucleotide polymorphisms (SNPs) of LTA4H as rs2072512, rs2540489, rs2540500 and rs6538697 were selected. Generalized estimating equation (GEE) was used to adjust for in-family correlation in the analysis of discordant sib pairs. Conditional logistic regression model and family based association test (FBAT) were both used to test the associations of LTA4H gene with ischemic stroke and its subtypes.

RESULTSIn total, data from 356 discordant sib pairs from 234 ischemic stroke patient pedigrees were analyzed. Results of GEE showed that hypertension, diabetes mellitus and hyperlipoidemia were associated with ischemic stroke. According to the association test results, rs2540489 G allele was found to be associated with ischemic stroke, both in the additive model (OR = 0.62, 95% CI: 0.41-0.94) and in the recessive model (OR = 0.48, 95% CI: 0.23-1.02). For two main ischemic stroke subtypes, rs2540489 G allele was found to be associated with large-artery atherosclerosis stroke in the additive model (OR = 0.48, 95% CI: 0.24-0.95) and in the recessive model (OR = 0.25, 95% CI: 0.07-0.93). After adjusting age, sex and other factors, the associations mentioned above, still existed.

CONCLUSIONrs2540489 polymorphism in LTA4H gene seemed to be associated with large-artery atherosclerosis stroke.

Alleles ; Atherosclerosis ; Brain Ischemia ; genetics ; Diabetes Mellitus ; Epoxide Hydrolases ; genetics ; Humans ; Hyperlipidemias ; Hypertension ; Logistic Models ; Pedigree ; Polymorphism, Single Nucleotide ; Siblings ; Stroke ; genetics

BACKGROUNDInflammation is a major cause of restenosis after coronary stenting. Intercellular adhesion molecule-1 (ICAM-1) is an important adhesion molecule that plays a key role in the tight adhesion between leukocytes and vascular endothelium. The object of this study was to investigate the association between the K469E polymorphism of the ICAM-1 gene and restenosis after coronary stenting in North Chinese population.

METHODSThe ICAM-1 K469E polymorphism was genotyped using polymerase chain reaction-restriction fragment length polymorphism method in 124 patients who had undergone coronary stenting and coronary angiography at least 3 months earlier. Information on clinical risk factors and procedure-related data were also collected.

RESULTSOf 124 enrolled patients in total, there were 72 cases of in-stent restenosis. The restenosis rate in this population was 58.1%. The frequencies of the three possible genotypes of the ICAM-1 K469E polymorphism were: KK genotype 50.8%, EE genotype 41.9%, and EK genotype 41.9%. Among restenosis patients, the frequency of the KK genotype was 58.3% and the frequency of E allele carriers was 41.7%. Among non-restenosis patients, the frequency of the KK genotype was 40.4%, and the frequency of E allele carriers was 59.6%. The distribution of these two genotype groups between restenosis and non-restenosis patients was significantly different (P = 0.049). Using multivariate logistic regression, the difference between the two groups was more apparent. The odds ratio of KK homozygotes vs E allele carriers was 2.6, with 95% confidence interval 1.2 - 5.8 (P = 0.018). After grading of risk factors, we found that the KK genotype was a stronger predictor of in-stent restenosis in obesity or hyperlipemia patients, with an odds ratio of 9.3 and 3.7, respectively (P < 0.05).

CONCLUSIONIn our study population, KK homozygotes of the ICAM-1 codon 469 mutation had a higher risk of restenosis after coronary stenting, especially in the case of obese or hyperlipemia patients.

Asian Continental Ancestry Group ; genetics ; China ; Codon ; Coronary Restenosis ; genetics ; Female ; Genotype ; Humans ; Hyperlipidemias ; complications ; Intercellular Adhesion Molecule-1 ; genetics ; Male ; Middle Aged ; Obesity ; complications ; Polymorphism, Genetic ; Stents

OBJECTIVETo study the relationship between the polymorphism of ApoE gene and TCM syndrome type of primary hyperlipemia.

METHODSApoE genotype of 102 patients with hyperlipemia was detected by gene PCR sequencing.

RESULTSA total of five genotypes were detectable, they were E2/2, E3/3, E4/4, E2/3 and E3/4. The frequency of E3/4 + E4/4 and epsilon4 allelotype detected in the patients of Gan-Shen Yin deficiency syndrome type were significantly higher than those in patients of Pi-Shen Yang-deficiency type or of phlegm stagnation type (P < 0.05, P < 0.01), and which in patients of Qi-stagnation caused blood stasis type were significantly higher than those in patients of phlegm stagnation type ( P < 0.05).

CONCLUSIONPolymorphism of ApoE gene is related in a certain degree to TCM syndrome type of primary hyperlipemia.

Adult ; Aged ; Aged, 80 and over ; Apolipoproteins E ; genetics ; Diagnosis, Differential ; Female ; Genotype ; Humans ; Hyperlipidemias ; diagnosis ; genetics ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Polymorphism, Genetic ; Yin Deficiency ; genetics

Humans ; Diabetes Mellitus, Type 2/genetics* ; Hyperlipidemias ; Mutation ; Insulin Resistance/genetics* ; Metabolic Diseases ; Proto-Oncogene Proteins c-akt ; Perilipin-1/genetics*