Schisantherin A alleviated alcohol-induced liver injury by the regulation of alcohol metabolism and NF-kB pathway.

Schisantherin A (SinA), one of the most abundant active ingredients of Schisandra chinensis, was reported to protect and benefit the liver, however, its effect on alcohol-induced liver injury (ALI) was still not clear. In the present study, an ALI mice model was induced by feeding mice an alcohol-containing liquid diet for four weeks. Then, 100 mg/kg or 200 mg/kg SinA was administered to mice every day by gavage for the last two weeks. Histopathological analysis showed that alcohol-induced liver lipid vacuoles were reduced by SinA. The activities of aspartate aminotransferase (AST, 61.90 ± 14.65 vs. 93.65 ± 20.50, 50.46 ± 13.21 vs. 93.65 ± 20.50) and alanine transaminase (ALT, 41.29 ± 9.20 vs. 64.04 ± 18.13, 36.52 ± 7.71 vs. 64.04 ± 18.13) in the serum of ALI mice were significantly reduced by 100 mg/kg or 200 mg/kg SinA when compared with control mice. Alcohol-induced oxidative stress and the inflammatory response in the liver were suppressed by SinA in a dose-dependent manner. Meanwhile, treatment with SinA decreased alcohol dehydrogenase (ADH) activity and increased acetaldehyde dehydrogenase (ALDH) activity in ALI mice. Alcohol-induced upregulation of CYP2E1 and CYP1A2 in the liver was inhibited by SinA. Further, SinA suppressed activation of the NF-kB pathway in ALI mice. In conclusion, our findings demonstrate that SinA is able to protect against ALI, and this may be, at least in part, caused by regulation of alcohol metabolism and the NF-kB pathway. Our data suggest a therapeutic potential of SinA in the treatment of ALI.

The activity of class I, II, III and IV alcohol dehydrogenase isoenzymes and aldehyde dehydrogenase in the sera of bladder cancer patients.

OBJECTIVES: Studies on alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activity in the sera of patients with malignant neoplasms show that cancer cells in many organs may release ADH isoenzymes into the blood. The aim of this study was to investigate the differences in the activity of ADH isoenzymes and ALDH in the sera of patients with bladder cancer (BCa), and with different grades of the disease. MATERIAL AND METHODS: Blood samples were taken from 39 patients with BCa (15 patients with low-grade and 24 with high-grade BCa) and from 60 healthy subjects. Class III and IV of ADH and total ADH activity were measured using the photometric method, while class I and II ADH and ALDH activity using the fluorometric method with class-specific fluorogenic substrates. RESULTS: The activity of the class I ADH isoenzyme and total ADH was significantly higher in the sera of BCa patients as compared to control group. Analysis of ALDH activity did not show statistically significant differences between the tested groups. Significantly higher total activity of ADH in comparison to control was found in both, low-grade and high-grade BCa group. The activity of ADH class I was also significantly higher in high-grade BCa group when compared to low-grade patients and controls. CONCLUSION: The increase of total ADH activity in the sera of BCa patients seems to be caused by isoenzymes released from cancerous cells. The higher activity of ADH I probably resulted from metastatic tumors as significant increase was detected only in the sera of high-grade bladder cancer patients.

Identification of 4-Trimethylaminobutyraldehyde Dehydrogenase (TMABA-DH) as a Candidate Serum Autoantibody Target for Kawasaki Disease.

Kawasaki disease (KD), an acute vasculitis that preferentially affects coronary arteries, is still the leading cause of acquired heart disease in children. Although the involvement of immune system malfunction in the onset of KD is suggested, its etiology still remains to be clarified. We investigated autoantibodies in KD patients, which are frequently found in sera from patients with autoimmune diseases, vasculitides and arteritides. We performed two-dimensional western blotting and LC-MS/MS to analyze the antigens of autoantibodies, detected two protein spots with 4 out of 24 sera from KD patients but not with 6 control sera, and identified the antigens as 4-trimethylaminobutyraldehyde dehydrogenase (TMABA-DH). A slot blot analysis with TMABA-DH as an antigen also revealed higher reactivities of patients' sera than control sera (positive rates: 18/43 vs 3/41). Using an enzyme-linked immunosorbent assay (ELISA), we found that the reactivity of anti-TMABA-DH antibodies in sera from KD patients was significantly higher than that in sera from age-matched controls. The optimal cut-off value of 0.043 had a sensitivity of 83.7% and a specificity of 80.0% in detecting KD patients (positive rates: 37/43 for KD patients, 9/41 for controls). Immunohistochemistry performed on thin sections of rat heart revealed that TMABA-DH colocalized with myosin light chains in cardiac myocytes. Patient sera with high reactivity gave similar immunostaining pattern. These results suggest that the detection of anti-TMABA-DH autoantibody could be a potential strategy for a diagnosis of KD.

Alcohol dehydrogenase (ADH) isoenzymes and aldehyde dehydrogenase (ALDH) activity in the sera of patients with gastric cancer.

BACKGROUND: Investigations have shown that alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) are present in some cancer cells and can play role in carcinogenesis. In recent experiments we found elevated alcohol dehydrogenase class IV activity in gastric cancer cells. This suggests these changes may be reflected by enzyme activity in the serum. AIM: In this work we measured the activity of alcohol dehydrogenase isoenzymes and aldehyde dehydrogenase in the sera of patients with gastric cancer matched on gender. METHODS: Serum samples were taken for routine biochemical investigation from 55 patients with gastric cancer, before treatment, and from 55 control subjects. Total ADH activity was measured photometrically and ALDH activity by a fluorimetric method. For measurement of the activity of class I isoenzymes we used a fluorimetric method, with class-specific fluorogenic substrates. The activity of class III and IV alcohol dehydrogenase was measured photometrically. RESULTS: The activity of the class IV ADH isoenzyme was significantly higher in the sera of patients with gastric cancer. The median activity of this isoenzyme in the total cancer group was approximately 47% higher (7.45 mU/l) than the control level (5.08 mU/l). For this reason total ADH activity was also significantly increased. The activities of other tested ADH isoenzymes and ALDH were unchanged. CONCLUSION: Changes in the activity of, especially, class IV ADH in the sera of patients with gastric cancer seems to be caused by release of the isoenzyme from cancer cells.

Alcohol dehydrogenase (ADH) isoenzymes and aldehyde dehydrogenase (ALDH) activity in the sera of patients with pancreatic cancer.

BACKGROUND: Numerous experiments have shown that alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) are present in cells of various cancer and can play role in carcinogenesis. In previous investigations we have found elevated levels of alcohol dehydrogenase class III activity in pancreatic cancer cells. It can suggest that these changes may be reflected by enzyme activity in the serum. AIM: In this paper we have measured the activity of alcohol dehydrogenase isoenzymes, and aldehyde dehydrogenase in the sera of patients with pancreatic cancer. METHODS: Serum samples were taken for routine biochemical investigation from 56 patients with pancreatic cancer before treatment. Total ADH activity was measured by photometric method with p-nitrosodimethylaniline (NDMA) as a substrate, and ALDH activity by the fluorometric method with 6-methoxy-2-naphtaldehyde as a substrate. For the measurement of the activity of class I isoenzymes, we employed the fluorometric methods, with class-specific fluorogenic substrates. The activity of class III alcohol dehydrogenase was measured by the photometric method with n-octanol and class IV with m-nitrobenzaldehyde as a substrate. RESULTS: A statistically significant increase of class III alcohol dehydrogenase isoenzymes was found in the sera of cancer patients. The median activity of this class isoenzyme in the total cancer group increased about 22% (13.52 mU/l) in the comparison with the control level (11.08 mU/l). The total alcohol dehydrogenase activity was significantly higher (19.7%) among patients with cancer than healthy ones. The activities of other tested ADH isoenzymes and total ALDH were unchanged. The activity of the class I ADH isoenzyme was significantly higher in the sera of drinkers with pancreatic cancer than non-drinkers. CONCLUSION: The increased total activity of alcohol dehydrogenase and class III isoenzyme in the sera of patients with pancreatic cancer can be caused by release of this isoenzyme from cancer cells.

Effects of acute acetaldehyde, chronic ethanol, and pargyline treatment on agonist responses of the rat aorta.

The purpose of this investigation was to determine the mechanism(s) underlying the vasorelaxant effects of acute versus chronic acetaldehyde (ACA) exposure, in particular, the role of intact endothelium on contractile responses of rat aortic ring segments to potassium chloride (KCl) or norepinephrine (NE). The acute effects of ACA were assessed in preparations from normal animals maintained on a standard rat chow (non-ethanol-ingesting). The monoamine oxidase inhibitor, pargyline, elevates plasma ACA levels by decreasing acetaldehyde dehydrogenase activity. Accordingly, preparations from pargyline-treated ethanol-ingesting animals (PE) were used to assess the effects of chronic (12 weeks) ACA and results were compared to those of pargyline-treated non-ethanol-ingesting (P) rats fed a standard liquid diet. In normal and P preparations, maximal inotropic response to KCl and NE were greater in endothelium-denuded than in endothelium-intact preparations. However, in aortic rings obtained from PE rats, the maximal inotropic response to KCl was depressed only in endothelium-denuded rings and that to NE was nearly identical in endothelium-denuded compared to endothelium-intact preparations. Acute ACA (30 mM) exposure significantly reduced both NE- and KCl-induced contractile responses in muscles from all groups. The magnitude of the vasorelaxant effect of this [ACA] on NE-induced responses was endothelium-independent and was similar between groups. However, the vasorelaxant effect of ACA (30 mM) on KCl-induced contractile responses was significantly attenuated in muscles from PE animals with greater inhibition occurring in endothelium-denuded preparations. These results suggest that chronic acetaldehyde exposure leads to an impairment in the inotropic response to membrane depolarization in endothelium-denuded preparations resulting in depressed responsiveness. In addition, the acute vasorelaxant effect of ACA on KCl-induced contractures is significantly attenuated in preparations chronically exposed to ACA which suggests a possible development of tolerance.

Comparative study of alcohol metabolism in stroke-prone spontaneously hypertensive rats and Wistar-Kyoto rats fed normal or low levels of dietary protein.

Our previous experiments have shown that the appetite or preference for alcohol is affected by the rat strain and nutritional status, such as dietary protein levels. To determine the affected factors in alcohol preference, the alcohol metabolism in SHRSP (stroke-prone spontaneously hypertensive rats) and WKY (Wistar-Kyoto) rats fed with the standard level (15%) or low level (5%) purified egg protein diet (PEP) was investigated. The animals were kept on the experimental diets for 4 weeks. After 12 h fasting, a 15% ethanol solution was given in a dose of 100 mg ethanol per 100 g body weight with a gastric probe to all animals and the blood ethanol and acetaldehyde levels were determined. Compared with 15% PEP diet-fed SHRSP, WKY showed higher levels of blood ethanol and acetaldehyde. Furthermore, the same results were also observed in SHRSP and WKY fed with 5% PEP diet. On the other hand, regardless of the rat strain, rats fed a low level protein diet showed higher blood ethanol and acetaldehyde levels. We also found that there was no significant change in alcohol dehydrogenase (ADH) activity and acetaldehyde dehydrogenase (ALDH) activity between SHRSP and WKY. However, both SHRSP and WKY fed a 15% PEP diet showed higher ADH and ALDH activity compared with rats fed the 5% PEP diet. These results suggested that the affected factors of preference for alcohol may be correlated with blood ethanol and acetaldehyde levels after alcohol intake.

Vitamin B6 metabolism and binding to proteins in the blood of alcoholic and nonalcoholic men.

Blood obtained from nonalcoholic and alcoholic subjects was incubated with 100 nM [3H]pyridoxine to study its uptake and metabolism by erythrocytes and the binding of vitamin B6 metabolites to proteins in plasma and erythrocytes. Erythrocytes of the alcoholics accumulated tritium faster than those of the controls; however, they contained the same total amount of tritiated compounds by 15 min. After incubation for 30 min, the erythrocytes had converted most of the pyridoxine to pyridoxal phosphate and pyridoxal. Pyridoxal-P remained in the erythrocytes, and approximately 40% of the pyridoxal diffused into the plasma. [3H]Pyridoxal and [3H]pyridoxal-P levels in the erythrocytes and plasma of the alcoholics were similar to those in the controls. However, dialyzed hemolysates of the alcoholics had more [3H]pyridoxal and a lower percentage of [3H]pyridoxal-P than those of the controls. The total concentration of plasma pyridoxal-P was lower in the alcoholics than in the controls and did not change upon incubation of whole blood with pyridoxine or upon dialysis. The erythrocytes of the alcoholics and controls had similar concentrations of pyridoxal-P that increased 2.5-fold upon incubation of whole blood with pyridoxine for 30 min and returned to the initial concentrations upon dialysis. The amount of [3H]pyridoxal and [3H]pyridoxal-P bound to protein was assessed by treating hemolysate and plasma samples with borohydride before dialysis. More 3H was bound to protein in the erythrocytes than in the plasma. The amount of protein-bound 3H in the erythrocytes of the alcoholics was lower than that of the controls, whereas the amount of protein-bound 3H in plasma was similar in both groups.(ABSTRACT TRUNCATED AT 250 WORDS)

A fluorimetric assay for succinic semialdehyde dehydrogenase activity suitable for prenatal diagnosis of the enzyme deficiency.

Succinic semialdehyde dehydrogenase (SSAD) is an enzyme involved in the turnover of the neurotransmitter 4-aminobutyrate (GABA). Deficiency of SSAD results in developmental delay, ataxia, seizures and 4-hydroxybutyric aciduria. We have developed a simple fluorimetric assay for the enzyme and applied it to measurement of SSAD activity in a range of cell types often used for prenatal and postnatal diagnosis of enzyme defects. Lymphocytes from children with SSAD deficiency were found to have < 5% of the activity found in lymphocytes from normal children. Heterozygotes are asymptomatic and have intermediate enzyme activities. Although SSAD activity has been detected previously in uncultured chorionic villi, we found that SSAD was not expressed in cultured chorionic villus cells nor in some fibroblast-like amniocytes from control fetuses. Lymphocytes from fetal blood and non-fibroblastic amniocytes have high SSAD activities, and should be suitable for prenatal diagnosis of SSAD deficiency.

A double-blind placebo controlled study of healthy volunteers given a subcutaneous disulfiram implantation.

A study was carried out to elicit information of the pharmacological effect of implanted disulfiram (DS). Healthy volunteers were randomized to either of two groups; six were given 1 g DS implant and six subjects placebo (PL) implants. Each participant was subjected to one intravenous ethanol challenge before, and six intravenous ethanol challenges as well as one oral ethanol challenge after the implantation. No clinical signs of disulfiram-ethanol reactions (DER) were observed. The intravenous challenges did not result in any significant differences between pre- and post-implantation values for blood acetaldehyde concentrations in the DS group, and these values were not significantly different from the corresponding values in the PL group. However, somewhat higher blood acetaldehyde concentrations were recorded after an oral ethanol dose (0.8 g/kg) in the DS group. In a longitudinal study with oral ethanol challenges after implanted DS, one of three healthy volunteers had a significant higher blood acetaldehyde concentration three weeks after the implantation, while no such tendency was found in any subject tested earlier or later in the post-implantation period. The blood acetaldehyde levels after oral challenges were, however, far too low to produce a DER.

Concentration of vitamin B6 and activities of enzymes of B6 metabolism in the blood of alcoholic and nonalcoholic men.

The purpose of this study was to compare concentrations of vitamin B6 compounds and the activities of enzymes that synthesize or catabolize pyridoxal 5'-phosphate in the plasma and erythrocytes of nonalcoholic and alcoholic subjects. Blood was obtained from male nonalcoholics and chronic alcoholics with minimal liver damage and normal hematology. Plasma, erythrocyte, and urinary B6 compounds were analyzed by high performance liquid chromatography, and pyridoxal phosphate was also measured enzymatically. Erythrocyte pyridoxine kinase and pyridoxine phosphate oxidase and erythrocyte and plasma pyridoxine phosphate phosphatases were assayed. Plasma pyridoxal phosphate concentration was significantly lower in the alcoholics (31.3 +/- 3.6 nmol/liter) than in the nonalcoholics (58.7 +/- 7.5 nmol/liter). The concentrations of the other B6 compounds in plasma, erythrocytes, and urine were not different in the two groups. Plasma alkaline pyridoxine phosphate phosphatase activity was significantly higher in the alcoholics (4.05 +/- 0.36 nmol/(h.mg] than in the nonalcoholics (3.01 +/- 0.18 nmol/(h.mg]. The activities of erythrocyte kinase, oxidase, and phosphatases were not significantly different in the two groups. The relationship of plasma pyridoxal phosphate concentration to its metabolites and the activities of the enzymes involved in its metabolism was determined. Plasma pyridoxine phosphate phosphatase activity assayed at pH 9.0 or 7.4 correlated negatively with plasma pyridoxal phosphate concentration. The low pyridoxal phosphate concentration observed in the plasma of the alcoholic subjects may in part be related to increased plasma phosphatase activity.

Oxidation of [U-14C]succinic semialdehyde in cultured human lymphoblasts: measurement of residual succinic semialdehyde dehydrogenase activity in 11 patients with 4-hydroxybutyric aciduria.

The oxidation of [U-14C]succinic semialdehyde to 14CO2 has been investigated in cultured lymphoblasts to develop a whole cell assay for succinic semialdehyde dehydrogenase. We have previously demonstrated deficiency of this enzyme in extracts of white cells derived from 13 patients with 4-hydroxybutyric aciduria. Major goals were the demonstration of greater residual succinic semialdehyde dehydrogenase activity in patient cell lines and the better representation of physiology in vivo. In 18 control lymphoblast lines, the conversion of [U-14C]succinic semialdehyde to 14CO2 was 1579 +/- 310 dpm. The mean value in lymphoblasts derived from 11 patients with deficiency of succinic semialdehyde dehydrogenase was 112 +/- 36 dpm approximating 7% of the mean control value. Analysis of organic acids produced from [U-14C]succinic semialdehyde in control lymphoblasts indicated that 14CO2 emanated from the tricarboxylic acid cycle; the major metabolic products were succinic and lactic acids. In the presence of 5mM malonic and 2-propylpentanoic (valproic) acids, 14CO2 production in a control lymphoblast line was decreased by 68 and 45%, respectively. The whole cell assay is less laborious than our previously described assay employing cell extracts, and the general trend was the demonstration of higher residual levels of activity for lymphoblasts derived from patients.

Succinic semialdehyde dehydrogenase activity in bovine adrenal medulla and blood platelets: a comparative study with the brain enzyme.

In the present paper we report the presence of succinic semialdehyde dehydrogenase (SSADH) in bovine adrenal medulla and blood platelets. Both enzymes present some analogies with the brain enzyme in terms of cofactor requirements, optimal pH, mitochondrial localizaton and inhibition by AMP. However, the activity of the platelet enzyme is 100 times lower than that of the brain and affinities of both enzymes for their specific substrate succinic semialdehyde and NAD are different. The presence of SSADH in adrenal medulla and blood platelets allows us to confirm the presence of a complete GABA bypass in these tissues, where the neurotransmitter could have important regulator functions.

Multiple forms of formaldehyde dehydrogenase from human red blood cells.

Red cell hemolysates from nonrelated Finns were analyzed by electrofocusing on polyacrylamide gel, and formaldehyde dehydrogenase (EC 1.2.1.1) was located by an activity-staining method. Three forms of the enzyme were constantly found for all the individuals studied but no variants were observed in this population (n = 217). Human liver also had three formaldehyde dehydrogenase forms with locations identical to those of the red cell formaldehyde dehydrogenase. Population genetic studies of formaldehyde dehydrogenase can easily be performed with red cell hemolysates with the techniques described here, and there is no need to use liver biopsy samples.

Biochemical markers of chronic alcoholism.

Usefulness of several biochemical markers for the monitoring of chronic alcoholism were studied. Among generally used markers, only gamma-GTP showed a significant difference between alcoholic and non-alcoholic liver diseases. Serum glutamate dehydrogenase (GDH) activity was significantly high in alcoholic liver disease. When the ratios of GDH to ornithine carbamyl transferase (OCT) were calculated, differences between alcoholic and non-alcoholic liver diseases became clearer without overlapping of any value. Serum desialo-transferrin was found in about 60% of the alcoholics, and disappeared by abstinence. Microheterogeneity of serum protein was also found in other glycoproteins. Serum prealbumin level was significantly high in alcoholics without severe liver disease. Acetaldehyde dehydrogenase (ALDH) activity of erythrocytes was significantly low in alcoholics, and gradually increased after abstinence. These results indicate that microheterogeneity of glycoproteins, serum prealbumin level and erythrocyte ALDH activity are good markers of alcohol abuse, and serum GDH/OCT ratio is the most sensitive marker of alcoholic liver injury. Serum gamma-GTP activity is a good marker of both conditions.

Cefmetazole: clinical evaluation of efficacy and safety in Japan.

During the period February 1980-March 1982, clinical evaluation was carried out on 23,855 cases given cefmetazole (CMZ) in 3,916 medical treatment centres in Japan. The drug was found to have superior efficacy and to be of value for all age groups, ranging from infants and small children to the elderly, in infections due to Gram-positive cocci, Gram-negative bacilli and anaerobic bacteria sensitive to this drug. Experiments were also conducted to elucidate the mechanism of development of the disulfiram-like reaction found in cephems with a methyltetrazolylthiomethyl group at the 3 position, i.e., cefmetozole (CMZ), cefoperazone (CPZ) and latamoxef (LMOX). These experiments provided clear evidence that the reaction is due to a rise in the blood concentration of acetaldehyde, as a result of inhibition of acetaldehyde dehydrogenase activity caused by these cephems. The extent of increase in the blood concentration of AcH is proportional to the urinary excretion rate of mercaptomethyltetrazole (Me-TZ), being in the order CPZ greater than LMOX greater than CMZ. This order is believed to be due to the extent of distribution in bile by various antibiotics and to their stability in the tissue fluids.

Demonstration of 4-aminobutyric acid aminotransferase deficiency in lymphocytes and lymphoblasts.

Lysates of lymphocytes, isolated from whole blood, and Epstein-Barr virus transformed cultured lymphoblasts catalysed the transamination of 4-aminobutyric acid with 2-oxoglutaric acid as co-substrate. 4-Aminobutyric acid aminotransferase activity in lymphocyte and lymphoblast sonicates derived from 12 unrelated control individuals (6 each) was 39 +/- 19 pmol min(-1) (mg protein (-1] (mean +/- 1 SD). Activities in lysates of both types of cell derived from a Flemish patient were less than 3% of control. 4-Aminobutyric acid aminotransferase activity in sonicates derived from the parents and a healthy sibling were 15-37% of the control mean for lymphocytes and 13-20% of the control mean in lymphoblasts, respectively. Km values in a control lymphoblast sonicate were 0.63 and 0.08 mmol L(-1) for 4-aminobutyric and 2-oxoglutaric acids, respectively. These data indicate that the parents and healthy sibling are heterozygous and the patient is homozygous for a defective gene responsible for 4-aminobutyric acid aminotransferase deficiency, and that inheritance is autosomal recessive.

Effect of alcohol consumption on hepatic aldehyde dehydrogenase activity in alcoholic patients.

Two liver biopsies were performed 6 months apart in each of 29 patients with alcoholic liver disease. Hepatic aldehyde dehydrogenase activity was measured on each occasion. The patients were seen regularly and their alcohol consumption was assessed independently. Hepatic aldehyde dehydrogenase activity was unchanged in 17 patients who continued to drink to excess; it rose in 10 patients who significantly reduced their alcohol intake; and it fell dramatically in 2 patients who were virtually abstinent initially, but then began drinking heavily. These results clearly demonstrate that alcohol consumption itself depresses hepatic aldehyde dehydrogenase activity. It is unlikely that the low hepatic aldehyde dehydrogenase activity reported in alcoholics represents a primary abnormality predisposing to alcoholism or alcoholic liver disease.