This study aimed to identify the type of carnitine palmitoyltransferase 2 (CPT2) gene mutation in the child with carnitine palmitoyltransferase II (CPT II) deficiency and her parents and to provide the genetic counseling and prenatal diagnosis for the family members. As the proband, a 3-month-old female baby was admitted to the hospital due to fever which had lasted for 8 hours. Tandem mass spectrometric analysis for blood showed an elevated plasma level of acylcarnitine, which suggested CPT II deficiency. The genomic DNA was extracted from peripheral blood of the patient and her parents. Five exon coding regions and some intron regions at the exon/intron boundaries of the CPT2 gene were analyzed by PCR and Sanger sequencing. Amniotic fluid was taken from the mother during the second trimester, and DNA was extracted to analyze the type of CPT2 gene mutation. Sanger sequencing results showed that two mutations were identified in the CPT2 gene of the proband: c.886C>T (p.R296X) and c.1148T>A (p.F383Y), which were inherited from the parents; the second child of the mother inherited the mutation of c.886C>T (p.R296X) and showed normal acylcarnitine spectrum and normal development after birth. It is concluded that the analysis of CPT2 gene mutations in the family suggested that the proband died of CPT II deficiency and that the identification of the mutations was helpful in prenatal diagnosis in the second pregnancy.
Carnitine O-Palmitoyltransferase ; deficiency ; genetics ; Female ; Humans ; Infant ; Metabolism, Inborn Errors ; diagnosis ; genetics ; Mutation ; Prenatal Diagnosis

OBJECTIVE: To report on the clinical, metabolic and genetic characteristics of a child with carnitine palmitoyl transferase 1A (CPT1A) deficiency. METHODS: Clinical data and the level of acylcarnitine for a child who initially presented as epilepsy were analyzed. Genomic DNA was extracted from peripheral blood samples of the child and her parents and subjected to next-generation sequencing (NGS). RESULTS: Mass spectrometry of blood acylcarnitine indicated increased carnitine 0 (C0) and significantly increased C0/ (C16+C18). DNA sequencing revealed that the child has carried compound heterozygous variants of the CPT1A gene, namely c.1846G>A and c.2201T>C, which were respectively inherited from her mother and father. CONCLUSION CPT1A presenting initially as epilepsy was unreported previously. Analysis of blood acylcarnitine C0 and C0/ (C16 + C18) ratio and NGS are necessary for the identification and diagnosis of CPT1A deficiency. The c.1846G>A and c.2201T>C variants of the CPT1A gene probably underlay the disease in this child. Above finding has also enriched the spectrum of CPT1A gene variants.
Carnitine/blood* ; Carnitine O-Palmitoyltransferase/genetics* ; Child ; DNA Mutational Analysis ; Female ; Humans ; Hypoglycemia/genetics* ; Lipid Metabolism, Inborn Errors/genetics*

OBJECTIVETo analyze the clinical and molecular features of a child with carnitine palmitoyltransferase 1A (CPT1A) deficiency.

METHODSClinical data of the child was collected. Blood acylcarnitine was determined with tandem mass spectrometry. DNA was extracted from the child and his parents. All exons and flanking regions of the CPT1A gene were analyzed by PCR and Sanger sequencing.

RESULTSAnalysis showed that the patient carried compound heterozygous mutations c.1787T>C and c.2201T>C of the CPT1A gene, which derived his father and mother, respectively. Both mutations were verified as novel through the retrieval of dbSNP, HGMD and 1000 genome databases. Bioinformatic analysis suggested that the mutations can affect protein function.

CONCLUSIONAcyl carnitine analysis has been the main method for the diagnosis of CPT1A deficiency. The c.1787T>C and c.2201T>C mutations of the CPT1A gene probably underlie the disease in this patient. Gene testing can provide important clues for genetic counseling and prenatal diagnosis.

Base Sequence ; Carnitine O-Palmitoyltransferase ; deficiency ; genetics ; Exons ; Female ; Humans ; Hypoglycemia ; enzymology ; genetics ; Infant ; Lipid Metabolism, Inborn Errors ; enzymology ; genetics ; Male ; Molecular Sequence Data ; Point Mutation ; Pregnancy

OBJECTIVETo investigate the therapeutic effect of Dachaihutang on the development of atherosclerosis (AS) in rabbits and its possible mechanism by detecting the expression level of carnitine patmitoyl transferase-1 (CPT-1) in vascular smooth muscle layer of atherosclerotic rabbits, and search the new way and evidence for AS cures.

METHODThirty six male New Zealand white rabbits were divided randomly into control group, model control group, simvastatin group and Chinese traditional medicine dachaihutang group. After 9 weeks and 20 weeks of treatment, serum total cholesterol (TC) and triglyceride (TG) and low-density lipoprotein (LDL) levels were examined. At the end of 25 th weeks, histological changes in ascending aorta were studied by HE staining and histomorphometric analysis. The gene expression of CPT-1 in vascular smooth muscle layer of thoracic aorta was detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR).

RESULTCompared with model control group, in dachaihutang group serum TC and TG and LDL levels attenuated. Pathomorphology indicated that intima and media (I + M) became thinned, and the ratios of the thickness of intima to media(I/M) and the area of intima to media (SI/SM) were decreased (P < 0.05). Aortic intimal proliferation in Dachaihutang group was associated with a marked increase in CPT-1 expression in vascular smooth muscle layer of thoracic aorta. Compared to simvastatin group, except TG value, other values were higher in Dachaihutang group, however, there were no significant differences between the two groups.

CONCLUSIONThese findings suggest that early treatment with Dachaihutang not only induces a significant regression of arterial lesions of high cholesterol diet rabbits, but also has a crucial inhibited genesis and development of atherosclerosis effect by up-regulating CPT-1 expression in vascular smooth muscle layer.

Animals ; Aorta ; cytology ; drug effects ; enzymology ; Atherosclerosis ; drug therapy ; enzymology ; genetics ; Carnitine O-Palmitoyltransferase ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression ; drug effects ; Humans ; Male ; Myocytes, Smooth Muscle ; drug effects ; enzymology ; Rabbits ; Random Allocation

BACKGROUNDCurrently it is unclear whether lipid accumulation occurs in a particular sequence and its relationship with whole body insulin resistance (IR). This study aimed to answer this question.

METHODSMale Sprague-Dawley (SD) rats were fed on a normal or a high-fat diet for 20 weeks. Serum triglycerides (TG), serum free fatty acids (FFA), fasting plasma glucose (FPG), and liver and skeletal muscle TG were measured. The glucose infusion rate (GIR) and mRNA levels of acetyl-CoA carboxylase (ACC) and carnitine palmitoyltransferase-1 (CPT-1) in the liver and skeletal muscle were determined at different stages.

RESULTSCompared with rats fed on the normal diet, serum FFA was not significantly increased in rats fed on the high-fat diet until 20 weeks. In contrast, liver TG was significantly increased by the high-fat diet by four weeks (20-fold; P < 0.01), and remained elevated until the end of the study. However, skeletal muscle TG was not significantly increased by the high-fat diet until 20 weeks (10.6-fold; P < 0.01), and neither was the FPG. The GIR was significantly reduced (1.6-fold; P < 0.01) by the high-fat diet after 8 weeks. The mRNA levels of ACC gradually increased over time and CPT-1 decreased over time, in both the liver and skeletal muscle in rats fed the high-fat diet.

CONCLUSIONSLipid accumulation in the liver occurs earlier than lipid accumulation in the skeletal muscle. Fatty liver may be one of the early markers of whole body IR. Changes in the gene expression levels of ACC and CPT-1 may have important roles in the process of IR development.

Acetyl-CoA Carboxylase ; genetics ; Animals ; Blood Glucose ; analysis ; Carnitine O-Palmitoyltransferase ; genetics ; Fatty Acids, Nonesterified ; blood ; Fatty Liver ; etiology ; Insulin Resistance ; Lipid Metabolism ; Liver ; metabolism ; Male ; Muscle, Skeletal ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Triglycerides ; metabolism

OBJECTIVETo investigate the effects of maternal nutritional manipulation on fetal mRNA abundance of uncoupling protein UCP2, UCP3 and carnitine palmityl transferase 1 (CPT1), and find out an optimal maternal diet and targets for pharmacological prevention and treatment of obesity.

METHODSWistar pregnant rats were assigned to two groups which received a standard diet (SD) and a high protein diet (HPD) during pregnancy, respectively. After delivery, the male offspring were assigned to control group (CON) and high protein group (HP) according to their maternal diet, which were suckled by dams that received SD during pregnancy. Offspring were fed with SD from weaning (week 3) to week 8. Then CON were allocated to two groups: CON (SD during the whole experiment); HFCON (high fat control). HFCON and HP group rats were fed with high-fat diet (HFD) for 6 wk to induce obesity. At 0, 3, 8 and 14 wk of age, blood and tissue were collected for analyzing blood fat and abundance of UCP2, 3 and CPT1 mRNA.

RESULTSIn HP body weight and TG were decreased after weaning (F = 4.589, P = 0.039; F = 27.001, P = 0.000) and HFD (F = 16.076, P = 0.00; F = 71.518, P = 0.000). Obesity rates were significantly decreased in HP after HFD (chi2 = 8.076, P = 0.004). The abundance of UCP3 and CPT1 mRNA was persistently higher in HP than in CON or HFCON, and the abundance of UCP2 mRNA was also persistently higher than in CON or HFCON after weaning. Moreover the abundance of CPT1 mRNA was significantly increased after weaning and HFD compared with that after SD, the abundance of UCP2, UCP3 mRNA was also increased after HFD compared with that after SD.

CONCLUSIONSIncreasing protein intake during pregnancy might prevent offspring from HFD-induced obesity in adult, moreover might increase offspring the expression of UCP2, UCP3 and CPT1 mRNA. UCP2, UCP3 and CPT1 might participate in prevention and treatment of obesity by mediating fatty acid oxidation.

Animal Feed ; Animals ; Animals, Newborn ; Carnitine O-Palmitoyltransferase ; metabolism ; Dietary Proteins ; Female ; Fertile Period ; Ion Channels ; metabolism ; Male ; Mitochondrial Proteins ; metabolism ; Obesity ; metabolism ; Pregnancy ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Uncoupling Protein 2 ; Uncoupling Protein 3

OBJECTIVETo study the effect of Jinlida on changes in expression of skeletal muscle lipid transport enzymes in fat-induced insulin resistance ApoE -/- mice.

METHODEight male C57BL/6J mice were selected in the normal group (NF), 40 male ApoE -/- mice were fed for 16 weeks, divided into the model group (HF), the rosiglitazone group ( LGLT), the Jinlida low-dose group (JLDL), the Jinlida medium-dose group (JLDM), the Jinlida high-dose group (JLDH) and then orally given drugs for 8 weeks. The organization free fatty acids, BCA protein concentration determination methods were used to determine the skeletal muscle FFA content. The Real-time fluorescent quantitative reverse transcription PCR ( RT-PCR) and Western blot method were adopted to determine mRNA and protein expressions of mice fatty acids transposition enzyme (FAT/CD36), carnitine palm acyltransferase 1 (CPT1), peroxide proliferators-activated receptor α( PPAR α).

RESULTJinlida could decrease fasting blood glucose (FBG), cholesterol (TC), triglyceride (TG), free fatty acid (FFA) and fasting insulin (FIns) and raise insulin sensitive index (ISI) in mice to varying degrees. It could also up-regulate mRNA and protein expressions of CPT1 and PPARα, and down-regulate mRNA and protein levels of FAT/CD36.

CONCLUSIONJinlida can improve fat-induced insulin resistance ApoE -/- in mice by adjusting the changes in expression of skeletal muscle lipid transport enzymes.

Animals ; Apolipoproteins E ; deficiency ; genetics ; Blood Glucose ; metabolism ; CD36 Antigens ; genetics ; metabolism ; Carnitine O-Palmitoyltransferase ; genetics ; metabolism ; Dietary Fats ; adverse effects ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hypoglycemic Agents ; administration & dosage ; Insulin ; metabolism ; Insulin Resistance ; Lipid Metabolism ; drug effects ; Male ; Metabolic Diseases ; drug therapy ; enzymology ; genetics ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Muscle, Skeletal ; drug effects ; metabolism

OBJECTIVETo investigate the modulating effect on lipid and gene expressions of CPT I A caused by berberine (Ber) in experimental hyperlipidemia rats.

METHODMale SD rats were randomly divided into 5 groups according to the blood lipid values: normal group, hyperlipidemia group, 300 mg x kg(-1) x d(-1) Ber-treated group, 60 mg x kg(-1) x d(-1) Ber-treated group, and 7.2 mg x kg(-1) x d(-1) lovastatin-treated group. Normal group were fed with base diet and other groups were fed with high fat and cholesterol diet. 12 weeks after drugs were given the TC, TG, LDL-C, and HDL-C from rat blood samples were tested by automatic biochemistry analyzer. Gene expressions of CPT I A and PPARalpha were evaluated by RT-PCR and Western blot, respectively.

RESULTIt was shown that Ber significantly decreased TC and LDL-C, but increased HDL-C in dose-dependent manner, elevated expressions of CPT I A mRNA and protein without influence on PPARalpha expression. Similar effects from lovastatin on lipidemia were observed except the Ber effect on CPT I A gene expression.

CONCLUSIONBer has modulating effect on the lipid metabolism, the mechanism of which may be by promoting the CPT I A gene expression.

Animals ; Berberine ; administration & dosage ; Carnitine O-Palmitoyltransferase ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Gene Expression ; drug effects ; Humans ; Hyperlipidemias ; drug therapy ; enzymology ; genetics ; metabolism ; Lipid Metabolism ; drug effects ; Male ; PPAR gamma ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar