Carnosine (beta-Ala-L-His) has high antioxidant activity, and it is widely used in biology, chemical engineering, medicine and other fields. Its analogue syntheised in non-aqueous solvent and catalyzed by enzymes is high-effective but low-price, so it has great prospect. Here, we synthesized a carnosine analogue imidazole 4(5)-alanylamide-5(4)-carboxylic acid with imidazole-4,5-dicarboxylic acid and L-Alanine as substrates, alpha-chymotrypsin as catalyst in tetrahydrofuran (THF) solvent. Based on the orthogonal experiments, the optimized synthetic conditions are 4,5-dicarboxylic acid: L-alanine = 1:3 (m/m), alpha-chymotrypsin: substrates (4,5-dicarboxyl acid and L-alanine) = 1:200 (m/m), pH 8 phosphate buffer:THF = 1.6:10 (V/V), reaction temperature 35 degrees C, time 1.5 h. We separated the product with silica gel G60 thin-layer chromatography (TLC), and a new spot appeared at Rf (ratio to front) = 0.81; then the new spot was purified and characterized with UV spectra, high performance liquid chromatogram (HPLC) and 13C NMR (13C nuclear magnetic resonance). The UV spectra shows a new absorption peak at 310 nm, and the peak in 253 nm is largely strengthened; HPLC reserve times are all 4.5 min at 253 nm, 310 nm, 330 nm; 13C NMR shows 8 carbons. Combing with the catalytic mechanism of alpha-chymotrypsin, structure of the analogue is confirmed, i.e., imidazole 4(5)-alanylamide-5(4)-carboxylic acid.
Carnosine ; analogs & derivatives ; biosynthesis ; chemistry ; Catalysis ; Chromatography, High Pressure Liquid ; Chymotrypsin ; metabolism ; Furans ; chemistry ; Solvents

OBJECTIVETo investigate the protective effects of carnosine against experimental closed head injury (CHI) in mice.

METHODSThe CHI model was established by free-falling weight-drop. Carnosine (250 mg/kg or 500 mg/kg) was administered intraperitoneally 30 min before brain trauma, then q.d for 7 d; while normal saline was administrated for control group. The neurological defect was evaluated by neurological severity score (NSS) within 7 d; the survival rate and the histological alternations were observed.

RESULTSCarnosine prevented the body weight loss of mice at dose of 500 mg/kg; significantly increased the survival rate, and reduced the neurological defect and histological damage at dose of 250 and 500 mg/kg.

CONCLUSIONCarnosine can attenuate closed head injury in mice.

Animals ; Carnosine ; therapeutic use ; Disease Models, Animal ; Head Injuries, Closed ; drug therapy ; pathology ; Male ; Mice ; Mice, Inbred ICR

AIMClinical studies stated that low molecular weight compounds (< 1.0 kd) extracted from the new born calf liver could effectively inhibit the proliferation of tumor cells. In this report, we observed inhibition effects and their regulative mechanisms of taurine, ornithine, carnosine on the proliferation of HL-60 cells.

METHODSThree active ingredients, i.e., taurine, ornithine and carnosine were separated by ion-exchange chromatographic column and identified from the low molecular weight filtrate of new born calf liver. MTT assay was used to test the survival rate of HL-60 cells and normal lymphocytes treated by the three ingredients. The various effects of the three compounds on HL-60 cells were respectively evaluated by agarose gel electrophoresis, ESR and immunohistochemical methods.

RESULTSThese compounds effectively inhibited the proliferation of HL-60 cells and induced apoptosis which was determined by apoptotic changes in morphology and nuclear DNA degradation. Whereas no inhibition effects on normal lymphocytes were observed. In addition, the results of ESR showed that the activity of oxygen radical within HL-60 cells treated with there compounds decreased to trace level. Furthermore, in the immunohistochemical experiments, we found that the level of p45/skp2 in HL-60 cells decreased while the level of p27/kip increased.

CONCLUSIONThe taurine, ornithine and carnosine compounds can selectively suppress tumor cells proliferation by regulating the level of cell cycle proteins.

Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Carnosine ; pharmacology ; Cattle ; HL-60 Cells ; Humans ; Liver ; chemistry ; Ornithine ; pharmacology ; Taurine ; pharmacology

The effects of carnosine and related compounds (CRCs) including anserine, homocarnosine, histidine, and beta-alanine on monosaccharide autoxidation and H2O2 formation were investigated. The incubation of CRCs with D-glucose, D-glucosamine, and D, L-glyceraldehyde at 37degreeC increased the absorption maxima at 285 nm, 273 nm, and 290 ~ 330 nm, respectively. D, L-glyceraldehyde was the most reactive sugar with CRCs. The presence of copper strongly stimulated the reaction of carnosine and anserine with D-glucose or D-glucosamine. Carnosine and anserine stimulated H2O2 formation from D-glucose autoxidation in a dose-dependent manner in the presence of 10 muM Cu (II). The presence of human serum albumin (HSA) decreased their effect on H2O2 formation. Carnosine and anserine has a biphasic effect on alpha-ketoaldehyde formation from glucose autoxidation. CRCs inhibited glycation of HSA as determined by hydroxymethyl furfural, lysine residue with free epsilon-amino group, and fructosamine assay. These results suggest that CRCs may be protective against diabetic complications by reacting with sugars and protecting glycation of protein.
Absorption ; Anserine ; beta-Alanine ; Carbohydrates ; Carnosine* ; Copper ; Diabetes Complications ; Free Radicals ; Fructosamine ; Furaldehyde ; Glucose ; Histidine ; Humans ; Lysine ; Serum Albumin

Objective: To explore carnosine dipeptidase 1 (CNDP1) potential value as a diagnostic and prognostic evaluator of hepatocellular carcinoma (HCC). Methods: A gene chip and GO analysis were used to screen the candidate marker molecule CNDP1 for HCC diagnosis. 125 cases of HCC cancer tissues, 85 cases of paracancerous tissues, 125 cases of liver cirrhosis tissues, 32 cases of relatively normal liver tissue at the extreme end of hepatic hemangioma, 66 cases from serum samples of HCC, and 82 cases of non-HCC were collected. Real-time fluorescent quantitative PCR, immunohistochemistry, western blot, and enzyme-linked immunosorbent assay were used to detect the differences in mRNA and protein expression levels of CNDP1 in HCC tissue and serum. Receiver operating characteristic (ROC) curves and Kaplan-Meier survival were used to analyze and evaluate the value of CNDP1 in the diagnosis and prognosis of HCC patients. Results: The expression level of CNDP1 was significantly reduced in HCC cancer tissues. The levels of CNDP1 were significantly lower in the cancer tissues and serum of HCC patients than those in liver cirrhosis patients and normal controls. ROC curve analysis showed that the area under the curve of serum CNDP1 in the diagnosis of HCC patients was 0.753 2 (95% CI 0.676-0.830 5), and the sensitivity and specificity were 78.79% and 62.5%, respectively. The combined detection of serum CNDP1 and serum alpha-fetoprotein (AFP) significantly improved the diagnostic accuracy (AUC = 0.820 6, 95% CI 0.753 5-0.887 8). The diagnostic sensitivity and specificity of serum CNDP1 for AFP-negative HCC patients were 73.68% and 68.75% (AUC = 0.793 1, 95% CI 0.708 8-0.877 4), respectively. In addition, the level of serum CNDP1 distinguished small liver cancer (tumor diameter < 3 cm) (AUC = 0.757 1, 95% CI 0.637 4-0.876 8). Kaplan-Meier survival analysis showed that CNDP1 was associated with a poor prognosis in HCC patients. Conclusion: CNDP1 may be a potential biomarker for the diagnostic and prognostic evaluation of HCC, and it has certain complementarity with serum AFP.
Humans ; Carcinoma, Hepatocellular/genetics* ; Liver Neoplasms/pathology* ; Prognosis ; Carnosine ; alpha-Fetoproteins/metabolism* ; Biomarkers, Tumor/genetics* ; Liver Cirrhosis/diagnosis* ; ROC Curve

BACKGROUND: It is well known that neuronal degeneration can occur after a brief deprivation of energy source. To investigate whether glial astrocyte can induce a phenomenon of delayed cell death after transient energy loss and to see how different are the effects of nifedipine, lidocaine, carnosine and hypothermia on delayed toxicity in astrocyte. METHODS: Human astrocytoma cells (U1242MG) were used in this study. To assess the astrocyte survival during post-ischemic period after transient histotoxic hypoxia, 3-[4,5-dimethylthiazol-2yl]-2,5, diphenyl tetrazolium bromide (MTT) test was used. Compared to MTT test, tryphan blue test was also used to demonstrate membrane damage of affected cells. Studies on intracellular calcium dynamics during ischemic and post-ischemic period were carried out with fluo-3 and flow cytometry system. RESULTS: The percentage survival of astrocyte during post-ischemic period was decreasing with time. Calcium channel blocker nifedipine, sodium and calcium channel blocker lidocaine and free radical scavenger carnosine could not prevent post-ischemic cell damage. But, hypothermia was only an effective method in ameliorating post-ischemic cell death. Intracellular calcium increase during ischemia and post-ischemia was dependent on extracellular calcium influx. CONCLUSIONS: Only hypothermia was effective in reducing astrocyte death during post-ischemia after transient energy depletion. Intracellular calcium alterations during post-ischemia was from extracellular space.
Anoxia* ; Astrocytes* ; Astrocytoma ; Calcium ; Calcium Channels ; Carnosine ; Cell Death ; Extracellular Space ; Flow Cytometry ; Humans ; Hypothermia* ; Ischemia ; Lidocaine ; Membranes ; Neurons ; Nifedipine ; Sodium

The rapid increase in the prevalence of metabolic syndrome, which is associated with a state of elevated systemic oxidative stress and inflammation, is expected to cause future increases in the prevalence of diabetes and cardiovascular diseases. Oxidation of polyunsaturated fatty acids and sugars produces reactive carbonyl species, which, due to their electrophilic nature, react with the nucleophilic sites of certain amino acids. This leads to formation of protein adducts such as advanced glycoxidation/lipoxidation end products (AGEs/ALEs), resulting in cellular dysfunction. Therefore, an effective reactive carbonyl species and AGEs/ALEs sequestering agent may be able to prevent such cellular dysfunction. There is accumulating evidence that histidine containing dipeptides such as carnosine (beta-alanyl-L-histidine) and anserine (beta-alanyl-methyl-L-histidine) detoxify cytotoxic reactive carbonyls by forming unreactive adducts and are able to reverse glycated protein. In this review, 1) reaction mechanism of oxidative stress and certain chronic diseases, 2) interrelation between oxidative stress and inflammation, 3) effective reactive carbonyl species and AGEs/ALEs sequestering actions of histidine-dipeptides and their metabolism, 4) effects of carnosinase encoding gene on the effectiveness of histidine-dipeptides, and 5) protective effects of histidine-dipeptides against progression of metabolic syndrome are discussed. Overall, this review highlights the potential beneficial effects of histidine-dipeptides against metabolic syndrome. Randomized controlled human studies may provide essential information regarding whether histidine-dipeptides attenuate metabolic syndrome in humans.
Amino Acids ; Anserine ; Carbohydrates ; Cardiovascular Diseases ; Carnosine ; Chronic Disease ; Dipeptides ; Fatty Acids, Unsaturated ; Histidine ; Humans ; Inflammation ; Metabolism ; Oxidative Stress ; Prevalence ; Sequestering Agents

OBJECTIVETo investigate the effect of camosine on the injury of rat vascular endothelial cells(VECs) induced by hypoxia.

METHODSThe model of the injury of rat VECs induced by hypoxia was established. The effect of camosine on injury of VECs activity induced by hypoxia was determined by MTT assay. The levels of lactate dehydrogenase (LDH) activity in cell medium were measured with corresponding kit. The cell structure was observed under microscope after Coomassie brilliant blue R-250 staining.

RESULTSAfter culturing VECs with camosine (10 to 20 mmol/L) for 6 hours, the decrease in VECs activity induced by 12 and 24 hour hypoxia was inhibited. The release of LDH was also inhibited, and the integrity of cell structure remained.

CONCLUSIONCamosine has the protective effect on hypoxia injured VECs.

Animals ; Carnosine ; pharmacology ; Cell Hypoxia ; Cells, Cultured ; Endothelial Cells ; pathology ; Female ; Free Radical Scavengers ; pharmacology ; Male ; Protective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effect of alahistidine on brain histamine content and seizure development.

METHODSThe kindling seizure was induced by ip injection with subconvulsant dose of pentylenetetrazole every 48 h. Monoamines and their metabolites were measured using a HPLC system and fluorometric assay.

RESULTChronic low histamine feeding markedly decreased histamine content in cortex and hypothalamus, and promoted seizure development induced by pentylenetetrazole. However, alahistidine feed reversed the decreased histamine content and slowed seizure development caused by low histamine feed. Both low histamine and alahistidine feed had no effect on norepinephrine, dopamine and its metabolites.

CONCLUSIONAlahistidine may affect histaminergic system and seizure development.

Animals ; Brain Chemistry ; drug effects ; Carnosine ; analogs & derivatives ; pharmacology ; Histamine ; analysis ; Male ; Pentylenetetrazole ; Rats ; Rats, Sprague-Dawley ; Receptors, Histamine H1 ; physiology ; Seizures ; chemically induced

Animals ; Carnosine ; pharmacology ; therapeutic use ; Liver ; drug effects ; pathology ; Male ; Rats ; Rats, Wistar ; Reperfusion Injury ; prevention & control ; Shock, Hemorrhagic ; complications ; pathology