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Asian Pacific Journal of Tropical... Oct 2012To investigate and compare the inhibitory properties of free and bound phenolic extracts of clove bud against carbohydrate hydrolyzing enzymes (alpha-amylase &...
In vitro inhibition activity of polyphenol-rich extracts from Syzygium aromaticum (L.) Merr. & Perry (Clove) buds against carbohydrate hydrolyzing enzymes linked to type 2 diabetes and Fe(2+)-induced lipid peroxidation in rat pancreas.
OBJECTIVE
To investigate and compare the inhibitory properties of free and bound phenolic extracts of clove bud against carbohydrate hydrolyzing enzymes (alpha-amylase & alpha-glucosidase) and Fe(2+)-induced lipid peroxidation in rat pancreas in vitro.
METHODS
The free phenolics were extracted with 80% (v/v) acetone, while bound phenolics were extracted from the alkaline and acid hydrolyzed residue with ethyl acetate. Then, the interaction of the extracts with alpha-amylase and alpha-glucosidase was subsequently assessed. Thereafter, the total phenolic contents and antioxidant activities of the extracts were determined.
RESULTS
The result revealed that both extracts inhibited alpha-amylase and alpha-glucosidase in a dose-dependent manner. However, the alpha-glucosidase inhibitory activity of the extracts were significantly (P<0.05) higher than their alpha-amylase inhibitory activity. The free phenolics (31.67 mg/g) and flavonoid (17.28 mg/g) contents were significantly (P<0.05) higher than bound phenolic (23.52 mg/g) and flavonoid (13.70 mg/g) contents. Both extracts also exhibited high antioxidant activities as typified by their high reducing power, 1,1 diphenyl-2- picrylhydrazyl (DPPH) and 2, 2-azinobis-3-ethylbenzo-thiazoline-6-sulfonate (ABTS) radical scavenging abilities, as well as inhibition of Fe(2+)-induced lipid peroxidation in rat pancreas in vitro.
CONCLUSIONS
This study provides a biochemical rationale by which clove elicits therapeutic effect on type 2 diabetes.
Topics: Animals; Antioxidants; Carbohydrate Metabolism; Diabetes Mellitus, Type 2; Dose-Response Relationship, Drug; Enzyme Inhibitors; Ferrous Compounds; Flavonoids; Glycoside Hydrolase Inhibitors; Inhibitory Concentration 50; Lipid Peroxidation; Pancreas; Phenols; Plant Extracts; Polyphenols; Rats; Syzygium; alpha-Amylases; alpha-Glucosidases
PubMed: 23569846
DOI: 10.1016/S2221-1691(12)60228-7 -
Journal of Biochemistry May 1984DNase I in human urine was purified to an electrophoretically homogeneous state by column chromatographies on DEAE-lignocellulose, hydroxyapatite, DEAE-cellulose,... (Comparative Study)
Comparative Study
DNase I in human urine was purified to an electrophoretically homogeneous state by column chromatographies on DEAE-lignocellulose, hydroxyapatite, DEAE-cellulose, Sephadex G-75 and elastin-celite. The purified enzyme was immunologically identical with human pancreatic DNase I, but not with bovine pancreatic DNase I. The molecular weight and isoelectric point of the enzyme were estimated to be 4.1 X 10(4) and 3.6, respectively. The amino acid analysis revealed that 1 mol of the enzyme contained 8 mol of half-cystine. The N-terminal amino acid was identified as leucine by the dansyl chloride method. The enzyme was active in the presence of Mg2+, Co2+, or Mn2+, The optimum pH was around 6.5. The enzyme was stable in the pH range from 5.0 to 9.0 and at temperatures lower than 45 degrees C. The rate of hydrolysis of native DNA by the enzyme was twice as fast as that observed with heat-denatured DNA. This enzyme exhaustively degraded about 20% of the phosphodiester bonds in native DNA. The enzyme also degraded poly(dA) and poly(dT), but hardly degraded poly(dG) and poly(dC).
Topics: Amino Acids; Animals; Cations, Divalent; Cattle; Deoxyribonuclease I; Endodeoxyribonucleases; Epitopes; Humans; Hydrogen-Ion Concentration; Immunodiffusion; Kinetics; Molecular Weight; Pancreas; Species Specificity
PubMed: 6204972
DOI: 10.1093/oxfordjournals.jbchem.a134747 -
Gut Feb 1995The pathogenesis of alcoholic pancreatitis is not fully understood. An increase in pancreatic digestive and lysosomal enzyme synthesis because of ethanol consumption...
The pathogenesis of alcoholic pancreatitis is not fully understood. An increase in pancreatic digestive and lysosomal enzyme synthesis because of ethanol consumption could contribute to the development of pancreatic injury in alcoholics. This study aimed, firstly, to determine the effect of ethanol on the content and messenger RNA levels of pancreatic digestive enzymes and on the messenger RNA level of the lysosomal enzyme cathepsin B, and secondly, to examine the influence of concomitant protein deficiency (a known association of alcoholism and pancreatic injury) on these effects. A rat model of chronic ethanol administration was used in which rats were fed in groups of four, and for four weeks, protein sufficient and protein deficient diets with or without ethanol. Ethanol increased the pancreatic content of lipase but did not influence chymotrypsinogen or trypsinogen values. mRNA levels for lipase, trypsinogen, and chymotrypsinogen were raised in rats fed ethanol. Protein deficiency resulted in reduced tissue levels of lipase, chymotrypsinogen, and amylase but did not influence trypsinogen values. mRNA levels for proteases were increased in protein deficient rats, while those for lipase remained unaltered. Both ethanol and protein deficiency increased mRNA levels for cathepsin B. It is concluded that chronic ethanol consumption, in both protein sufficient and protein deficient states, increases the capacity of the pancreatic acinar cell to synthesise digestive and lysosomal enzymes.
Topics: Amylases; Animals; Blotting, Northern; Cathepsin B; Chymotrypsinogen; Ethanol; Lipase; Male; Pancreas; Protein Deficiency; RNA, Messenger; Rats; Rats, Sprague-Dawley; Trypsinogen
PubMed: 7533742
DOI: 10.1136/gut.36.2.287 -
Biochimica Et Biophysica Acta Apr 1998In the hypertrophic pancreas, we studied the oxidative degradation of polyamines, which are endogenous polycations important for cell division, growth and...
In the hypertrophic pancreas, we studied the oxidative degradation of polyamines, which are endogenous polycations important for cell division, growth and differentiation. To induce pancreatic hypertrophy, rats were fed on a semi-synthetic diet containing a daily dose of 42 mg phytohaemagglutinin per rat for 5 or 10 days. In the model, the activities of polyamine oxidase (the enzyme that degrades spermidine, spermine and mainly their acetyl derivatives) and diamine oxidase (the key enzyme of terminal catabolism of polyamines in vivo) increased by 100-180% and 90-100%, respectively, parallel to an elevation in polyamine content (40-100%). The results suggest that in pancreas hypertrophy, which does not exhibit stimulation of spermidine/spermine N1-acetyltransferase activity, increases in the activity of polyamine and diamine oxidases are related events that lead to putrescine formation and removal of excess polyamines.
Topics: Administration, Oral; Animals; Hypertrophy; Male; Organ Size; Oxidation-Reduction; Oxidoreductases Acting on CH-NH Group Donors; Pancreas; Phytohemagglutinins; Polyamines; Putrescine; Rats; Spermidine; Spermine; Substrate Specificity; Polyamine Oxidase
PubMed: 9630703
DOI: 10.1016/s0925-4439(98)00020-9 -
The Journal of Biological Chemistry Oct 2006Pancreatic acini release ATP in response to various stimuli, including cholecystokinin octapeptide (CCK-8), as we show in the present study. There were indications that...
Pancreatic acini release ATP in response to various stimuli, including cholecystokinin octapeptide (CCK-8), as we show in the present study. There were indications that pancreatic juice also contains enzymes that could hydrolyze ATP during its passage through the ductal system. The aim of this study was to determine which ATP-degrading and possibly ATP-generating enzymes were present in pancreatic secretion. For this purpose, pancreatic juice was collected from anesthetized rats stimulated with infusion of CCK-8. Purine-converting activities in juice samples were assayed by TLC using either [gamma-(32)P]ATP or (14)C/(3)H-labeled and unlabeled nucleotides as appropriate substrates. Data show that the juice contains the enzyme ecto-nucleoside triphosphate diphosphohydrolase that can hydrolyze both [(14)C]ATP and [(3)H]ADP about equally well, i.e. CD39. Reverse-phase high-performance liquid chromatography analysis additionally shows that this enzyme has broad substrate specificity toward other nucleotides, UTP, UDP, ITP, and IDP. In addition, secretion contains ecto-5'-nucleotidase, CD73, further converting [(3)H]AMP to adenosine. Along with highly active hydrolytic enzymes, there were also ATP-generating enzymes in pancreatic juice, adenylate kinase, and NDP kinase, capable of sequentially phosphorylating AMP via ADP to ATP. Activities of nonspecific phosphatases, nucleotide pyrophosphatase/phosphodiesterases, and adenosine deaminase were negligible. Taken together, CCK-8 stimulation of pancreas causes release of both ATP-consuming and ATP-generating enzymes into pancreatic juice. This newly discovered richness of secreted enzymes underscores the importance of purine signaling between acini and pancreatic ducts lumen and implies regulation of the purine-converting enzymes release.
Topics: 5'-Nucleotidase; Adenosine Triphosphate; Adenylate Kinase; Animals; Antigens, CD; Apyrase; Cells, Cultured; Female; Nucleoside-Diphosphate Kinase; Pancreas, Exocrine; Rats; Rats, Wistar
PubMed: 16885159
DOI: 10.1074/jbc.M602480200 -
Asian Pacific Journal of Tropical... Nov 2012To study the antioxidant efficacy of Commiphora mukul (C. mukul) gum resin ethanolic extract in streptozotocin (STZ) induced diabetic rats.
OBJECTIVE
To study the antioxidant efficacy of Commiphora mukul (C. mukul) gum resin ethanolic extract in streptozotocin (STZ) induced diabetic rats.
METHODS
THE MALE WISTAR ALBINO RATS WERE RANDOMLY DIVIDED INTO FOUR GROUPS OF EIGHT ANIMALS EACH: Control group (C), CM-treated control group (C+CMEE), Diabetic control group (D), CM- treated diabetic group (D+CMEE). Diabetes was induced by intraperitoneal injection of STZ (55 mg/kg/ bwt). After being confirmed the diabetic rats were treated with C. mukul gum resin ethanolic extract (CMEE) for 60 days. The biochemical estimations like antioxidant, oxidative stress marker enzymes and hepatic marker enzymes of tissues were performed.
RESULTS
The diabetic rats showed increased level of enzymatic activities aspartate aminotransaminase (AST), alanine aminotransaminase (ALT) in liver and kidney and oxidative markers like lipid peroxidation (LPO) and protein oxidation (PO) in pancreas and heart. Antioxidant enzyme activities were significantly decreased in the pancreas and heart compared to control group. Administration of CMEE (200 mg/kg bw) to diabetic rats for 60 days significantly reversed the above parameters towards normalcy.
CONCLUSIONS
In conclusion, our data indicate the preventive role of C. mukul against STZ-induced diabetic oxidative stress; hence this plant could be used as an adjuvant therapy for the prevention and/or management of diabetes and aggravated antioxidant status.
Topics: Alanine Transaminase; Animals; Antioxidants; Aspartate Aminotransferases; Commiphora; Diabetes Mellitus, Experimental; Heart; Lipid Peroxidation; Liver; Male; Myocardium; Oxidoreductases; Pancreas; Plant Extracts; Rats; Rats, Wistar
PubMed: 23569867
DOI: 10.1016/S2221-1691(12)60249-4 -
Journal of Biochemistry Jan 1996A novel trypsin-like serine proteinase was purified to homogeneity from the bovine pancreas microsome fraction. The enzyme was solubilized with...
A novel trypsin-like serine proteinase was purified to homogeneity from the bovine pancreas microsome fraction. The enzyme was solubilized with 3-[(3-cholamidopropyl)-dimethyl-ammonio]-1-propanesulfonate (CHAPS), and purified by a series of column chromatographic steps on Ultrogel AcA-34, trypsin inhibitor-Sepharose 4B, and arginine-Sepharose 4B. The molecular mass of this pancreas trypsin-like proteinase (bPTLP) was estimated to be 29.5 kDa by SDS-PAGE under reducing conditions. The NH2-terminal sequence of bPTLP is very homologous, but not identical to those of other serine proteinases, especially such as elastases IV, II, and III. Substrate specificity studies involving a synthetic substrate and glucagon indicated that the enzyme hydrolyzes Arg-X, Lys-X, and Leu-X bonds. The best synthetic substrate for bPTLP was t-butyloxycarbonyl Gln-Arg-Arg-4-methylcoumaryl 7-amide. The enzyme failed to hydrolyze the substrate for chymotrypsin and elastase. The enzyme activity was inhibited by diisopropyl fluorophosphate, p-amidinophenylmethane sulfonylfluoride, and leupeptin, indicating that it is a serine-proteinase. These findings show that bPTLP is a novel serine-proteinase which differs from all known proteinases. The physiological function of the enzyme has yet to be determined.
Topics: Amino Acid Sequence; Amino Acids; Animals; Cattle; Chemical Fractionation; Enzyme Stability; Glucagon; Humans; Hydrogen-Ion Concentration; Hydrolysis; Microsomes; Molecular Sequence Data; Pancreas; Sequence Homology, Amino Acid; Serine Endopeptidases; Serine Proteinase Inhibitors; Substrate Specificity
PubMed: 8907182
DOI: 10.1093/oxfordjournals.jbchem.a021193 -
Bioscience Reports Jan 2020The kinetic aspects of lipolysis by pulmonary phospholipase A2 (ChPLA2-V), chicken intestinal phospholipase A2 (ChPLA2-IIA) and chicken pancreatic phospholipase A2...
The kinetic aspects of lipolysis by pulmonary phospholipase A2 (ChPLA2-V), chicken intestinal phospholipase A2 (ChPLA2-IIA) and chicken pancreatic phospholipase A2 (ChPLA2-IB), from chicken have been compared using the monomolecular films technique, on short-chain phospholipids (with three different head groups) and on long-chain phospholipids. The main conclusions from our experimental data indicate that the maximum catalytic activities of ChPLA2-V on 1,2 phosphatidylcholine and 1,2 phosphatidylethanolamine reached 15.26 and 36.12 moles/cm2.min.mM, respectively, at a pressure of 15 and 35 dynes/cm, respectively. Whereas, those of ChPLA2-IB were 3.58 (at the pressure of 20 dynes/cm) and 4.9 moles/cm2.min.mM. However, hydrolysis of phosphatidylglycerol monolayers (C12PG), were very much higher compared with all the substrates tested with 122 moles/cm2.min. Surprisingly, the hydrolysis rate of ChPLA2-V on long-chain phosphatidylglycerol (C18PG) was very low (1.45 moles/cm2.min) compared with all tested substrates, even with the use of p-cyclodextrin. And thus, the fatty acid preference of ChPLA2-V was 2-decanoyl > 2-oleoyl with a PG head group. In order to gain significant correlations between enzyme's structures and their relative functions, we tried to examine the surface electrostatic potentials of the various secreted phospholipase 2 (sPLA2) from chicken. In the present study, we detailed that the substrate affinity, specificity and the hydrolysis rates of sPLA2 at each interface is governed by the surface electrostatic potentials and hydrophobic interactions operative at this surface.
Topics: Animals; Chickens; Fatty Acids; Hydrolysis; Intestines; Kinetics; Pancreas; Phosphatidylcholines; Phosphatidylethanolamines; Phospholipases A2; Phospholipids
PubMed: 31919493
DOI: 10.1042/BSR20192053 -
The Journal of Biological Chemistry Aug 1989Peptidylarginine deiminase (protein-L-arginine iminohydrolase, EC 3.5.3.15) is widely distributed in various organs of the mouse. Activity in salivary glands, pancreas,...
Peptidylarginine deiminase (protein-L-arginine iminohydrolase, EC 3.5.3.15) is widely distributed in various organs of the mouse. Activity in salivary glands, pancreas, and uterus is higher than that in the other organs. In submandibular gland and uterus, sex- and estrous cycle-related differences were observed, respectively. The activity in the submandibular gland from females was approximately four times higher than that in the male. In the uterus, the activity increased in proportion to the hyperplasia of the tissues. Peptidylarginine deiminase from the murine skeletal muscle resembles the enzyme obtained from other animal species with respect to enzymatic and chemical properties. Double immunodiffusion tests and immunoblotting analyses showed that the enzymes present in each murine tissue have the same molecular weight (81,000) and are immunologically indistinguishable. Immunohistochemical analyses of salivary glands and pancreas revealed an intense staining only of the exocrine cells. In uterus, the staining was restricted to the luminal and glandular epithelia of endometrium; the intensity of the staining changed during the course of the estrous cycle. Furthermore, immunoelectron microscopy showed that the enzyme is distributed diffusely in the cytoplasm of the exocrine cell. These observations indicate the general importance of peptidylarginine deiminase, presumably in a cytoplasmic secretory process of the exocrine cells.
Topics: Animals; Female; Hydrolases; Immunohistochemistry; Male; Mice; Pancreas; Protein-Arginine Deiminase Type 4; Protein-Arginine Deiminases; Subcellular Fractions; Submandibular Gland; Uterus
PubMed: 2753915
DOI: No ID Found -
Journal of Biochemistry Feb 1995Two endogenous inhibitors of nitric oxide synthesis, NG-monomethylarginine (MMA) and NG,NG-dimethylarginine (DMA), are broken down by NG,NG-dimethylarginine... (Comparative Study)
Comparative Study
Two endogenous inhibitors of nitric oxide synthesis, NG-monomethylarginine (MMA) and NG,NG-dimethylarginine (DMA), are broken down by NG,NG-dimethylarginine dimethylaminohydrolase (DDAH) [EC 3.5.3.18]. This enzymatic activity is widely distributed in rat tissues, and is correlated with the distribution of free MMA and DMA, and the production of nitric oxide. In this study, immunoblotting analyses with a monoclonal antibody directed towards rat kidney DDAH showed that immunoreactive proteins exist in human pancreas, kidney, and liver. Enzyme activity was definitely detected in these tissues, but the specific activity was lower than that in the corresponding rat tissue. These results are interesting in connection with the idea that the enzyme might play a role as a regulator of nitric oxide generation in human tissues.
Topics: Amidohydrolases; Animals; Antibodies, Monoclonal; Humans; Hydrolases; Immunoblotting; Kidney; Liver; Organ Specificity; Pancreas; Rats; Species Specificity; Substrate Specificity
PubMed: 7608105
DOI: 10.1093/jb/117.2.237