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Objective To analyze plasma vitamin E and CoQ10 levels in patients with autosomal recessive cerebellar ataxia for finding the evidence of the related pathogenesis research and therapeutic strategies.Methods The plasma vitamin E and CoQ10 levels were detected by high performance liquid chromatography (HPLC) with diode array detector in 123 probands of autosomal recessive cerebellar ataxia pedigrees.Quantitation was performed using vitamin E and CoQ10 external standard and two 5-point calibration curve;clinical manifestations were analyzed simuhaneously.Results Vitamin E and CoQ10 levels of healthy subjects in the plasma were (8.77 ± 2.28) μg/ml and (1.31 ± 0.38) μg/ml,respectively;the plasma vitamin E and CoQ10 levels of patients were (5.61 ± 2.04) μg/ml and (0.79 ± 0.26) μg/ml,respectively,which were significantly lower than those in healthy controls (t =11.87,13.15;all P＜ 0.01).Clinical manifestations were characterized by cerebellar symptoms,and gait instability was usually the first recognized abnormality.Most of early onset occurred before the age of 25 years (111/123);dysarthria and abnormal eye movement were observed,with cerebellar atrophy on MRI;concomitant symptoms were also present.Conclusions HPLC analysis shows that the plasma vitamin E and CoQ10 levels of patients with autosomal recessive cerebellar ataxia are generally lower than those in the healthy controls.Several patients with significant reductions in these two levels have genetic defects.The combination of clinical phenotypes,biochemical indexes and genetic analyses will be helpful for the establishment of diagnosis and specific treatment.
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Objective To study the effect of coenzymes complex combined with vitamin E on liver and kidney inju-ry induced by neonatal hyperbilirubinemia. Methods One hundred and fifty full-term neonatals with hyperbilirubinemia were chosen as observation groups, who were divided into mild, moderate and severe groups according bilirubin. Forty five healthy full-term newborns in the same period, who are either healthy or with physiological jaundice, were selected as con-trol group. Serum total bilirubin, gamma-glutamyl transferase (γ-GGT) , Malondialdehyde (MDA) and Cys-C were measured within 24 hours of hospital admission.The observation groups were randomly divided into regular and combined treatment groups. Coenzymes complex combined with vitamin E were given in addition to regular method to combined group while reg-ular group only received regular methods.All above biochemical indexes were tested in the 7th day after medication admin-istration. Results Serum MDA were higher in all observation groups (mild, moderate and severe groups) than in control group (P＜0.05);but the levels ofγ-GGT and Cys-C increased in moderate and severe groups compared with control group (P＜0.05). There were positive correlations (P＜0.05) between levels of serum total bilirubin with MDA,γ-GGT and Cys-C. Positive linear correlation were found between MDA with γ-GGT and Cys-C(P＜0.01). After early intervention,γ-GGT, Cys-C and MDA declined with drop of bilirubin level. This is more prominent and faster in a in combined treatment group than regular group (P＜0.05).Conclusion In hyperbilirubinemia newborns, lipid peroxidation activated by bilirubin may lead to damages of liver and kidney. Coenzymes combined with vitamin E have protective effect to these damages.
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Objective To explore the changes and clinical significance of plasma redox status in patients with acute myocardial infarction,angina pectoris and people with normal coronary artery.Methods According to the clinical manifestation,electrocardiograms and the myocardium markers,68 acute myocardial infarction patients (group A) were involved.68 angina pectoris patients (group B) and 68 normal coronary artery people (group C) were also chosen according to the coronary artery radiographs.Peripheral venous blood of 3 groups were collected.Plasma glutathione (reduced form GSH and oxidized form GSSG) and nicotinamide adenine dinucleotide phosphate (reduced form NADPH and oxidized form NADP+) were detected.The GSH/GSSG and NADPH/NADP+ redox potentials were calculated according to Nernst equation.Results Along with the pathological aggravation of coronary artery (from group C to group A),the levels of GSH [(8.39±1.03)μmol/L,(6.54±0.94) μmol/L,(4.49±0.86) μmol/L,respectively] and GSH/GSSG (14.22±2.14,9.76±1.67,5.76±1.18,respectively) were gradually reduced; the levels of GSSG [(0.59±0.03) μmol/L,(0.67±0.04) μmol/L,(0.78 ± 0.05) μmol/L,respectively] and GSH/GSSG redox potential [(-150.43±3.43) mV,(-141.22±3.12) mV,(-135.21±2.31) mV,respectively] were gradually increased (all P＜ 0.05); the changes of NADPH/NADP+ redox status were similar to GSH/GSSG but milder.Conclusions The imbalance of plasma redox status deviating to oxidation has a relationship with atheromatous plaque formation,plaque rupture and plaque thrombosis in the coronary artery.
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Objective: To establish a routine for the extraction of the total levels of CoQ10 in human plasma through the Ultra High Performance Liquid Chromatography (UHPLC). Methods: Two extraction protocols were tested: a) methanol: hexane and b) 1-propanol. The following parameters were analyzed: extraction temperature (19ºC and 4ºC), extraction tubes (glass and polypropylene), and surfactants (SDS, Triton X-100, Tween-20) at different concentrations, i.e., 1%, 3%, 5% and 10%. Results: The results showed that the method of extraction of CoQ10 in a sample of human plasma at 4ºC, using solvents methanol: hexane (85:15, v/v) in the presence of surfactant Tween-20 at 3% and polypropylene tubes showed better efficiency and reproducibility when compared to the method with 1-propanol. Conclusion: By the analyses performed, it was possible to observe that the addition of the surfactant Tween-20 promoted an increase in the recovery of CoQ10 by the methanol:hexane extraction method. This method showed good reproducibility, with a low coefficient of variation and high sensitivity, since CoQ10 was detected in samples of plasma of a control individual using a UV-type detector. The use of UHPLC equipment allowed a total analysis with total run time of 3.5 minutes, enabling the rapid achievement of results, considered mandatory for laboratory routines.
Objetivo: Estabelecer uma rotina de extração dos níveis totais de CoQ10 em plasma humano por meio da análise por Cromatografia Líquida de Ultra Alta Eficiência (UHPLC). Métodos: foram testados dois protocolos de extração: a) metanol:hexano e b) 1-propanol. Os seguintes parâmetros foram analisados: temperatura de extração (19ºC e 4ºC), tubos de extração (vidro e polipropileno), surfactantes (SDS, Triton X-100, Tween-20) em diferentes concentrações 1%, 3%, 5% e 10%. Resultados: Os resultados mostraram que o método de extração de CoQ10 em amostra de plasma humano, a 4ºC, utilizando-se os solventes metanol:hexano (85:15, v/v) na presença do surfactante Tween-20 a 3% e tubos de polipropileno apresentou melhor eficiência e reprodutibilidade quando comparado ao método com 1-propanol. Conclusão: A adição do surfactante Tween-20 no processo de preparação de amostra promoveu um aumento na recuperação da CoQ10 pelo método de extração metanol:hexano observada pela boa reprodutibilidade das prelicatas, pelo baixo coeficiente de variação e alta sensibilidade uma vez que a CoQ10 foi detectada em amostras de plasma de um indivíduo controle utilizando-se um detector do tipo UV. Além disso, a utilização de um equipamento de UHPLC proporcionou a obtenção de uma análise com tempo total de corrida de 3,5 minutos, o que viabiliza a obtenção rápida de resultados, considerado mandatório para rotinas laboratoriais.
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Chromatography, Liquid , Coenzymes , Surface-Active AgentsABSTRACT
Our study on the intracellular distribution of B12 and DBC on a normal and injured liver following the administration of these labelled test materials show that (1) with increasing interval, the pattern of distribution of these two closely related compounds in the various cellular fractions will simulate each other; (2) chromatography of B12 and DBC obtained from soluble fractions exhibited nearly identical Rf values; both findings lend support to our belief that the conversion of vitamin B12 to 5,6 dimethylbenzimidazole cobamide coenzyme took place; (3) in an injured liver, the initial uptake of either B12 or DBC activity is significantly different from the uptake, in almost all cellular fractions, of an intact liver; however, with prolonged interval, the pattern of distribution of B12 and DBC among the individual fractions will eventually be identical. (Summary)
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Objective To study the effect on bone formation by intermittent administration of simvastatin. Methods Seventy three month old virgin female rats, weighted (200?10) g each had been divided into two groups ovariectomized (50 rats) and pseudo ovarectomized (20 rats). Ten rats each of Group A and Group B were sacrified at the 30 th day for the confirmation of successful establishment models of the osteoporosis. Then the remaining animals were re divided into 5 groups: Group A, pseudo ovarectomized control, 10 rats; Group B, ovarectomized control, 10 rats; Group C, given Nylestriol 0.1 mg/(kg?d), 10 rats; Group D, 10 rats, given calcium 10 mg and vitamin D3 2 IU/(kg?d); Group E, 10 rats given simvastatin 5 mg/(kg?d) for 14 days; and the drug was suspended for 28 days, then simvastatin was given for another 14 days. All of the animals were given the agents through gastric tube 30 days after surgery. At 120 th day all the rats were sacrified for the measurements of bone mineral content and bone mineral density by the dual energy X ray absorptiometry for bone mophometrical study. Results The average of bone mineral content was (46?11.34) mg (P
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The author investigated the function of coenzyme NADH in increasing the level of energy metabolism, repairing cellular damage, improving cellular stress ability, decreasing cytotoxicity of chemotherapy drugs and radiation against normal tissue. The molecular regulation mechanism of NADH in cytoprotection was elucidated. A new prevention and cure way was provided in cytoprotection treatment for clinical disease.
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To investigate the role of NADH in preventing apoptotic damage of human hepatic cells induced by chemotherapy drug cisplatin, the ultrastructural changes in hepatic cells were examined with transmission and scanning electron microscopy, propidium iodide (PI) staining was used to measure the apoptotic rate with flow cytometry, the expressions of p53 and bcl 2 gene were detected with RT PCR, the change in caspase 3 and caspase 8 of apoptotic molecules was examined with colorimetric assay. The results showed that, compared with the group of cisplatin, morphological apoptotic changes were not obvious, the apoptosis rate was significantly decreased, p53 mRNA expression was decreased, bcl 2 mRNA expression was increased, caspase 3 and caspase 8 activity levels were kept in a low level in the group of NADH /DDP. The study indicates that NADH can prevent apoptosis of human hepatic cells, and increase the possibility of using NADH to reduce side effects of chemotherapy.