1.Regulating function of coenzymization and decoenzymization of the lactate dehydrogenase isozymes in the mouse tissues during hypoxia.
Yuan MINGXIU ; Jiang HUI ; Deng AIPING ; Zhou QINGZHONG
Chinese Medical Sciences Journal 2003;18(2):128-131
OBJECTIVETo study the characteristics of changes of LDH enzyme patterns of mice under slight hypoxia.
METHODSMice treated with artificial hypoxia, various tissues were made for the test of LDH enzymatic activity by the specific staining technique. LDH (1-5) relative percentage enzymatic activity (RPEA) were measured with CS-910 dual-wavelength thin layer chromatography scanner.
RESULTSThe RPEA of LDH isozymes of various tissues after slight hypoxia shifted to the isozymes LDH1 and LDH2, whose principal subunits are H subunits, and the RPEA of LDH1 (H4), LDH2 (H3M) increased, while RPEA of LDH5 (M4) in various tissues decreased prominently except the cardiac muscle, and that of LDH4 (HM3) decreased as well. After polyacrylamide gel electrophoresis (PAGE) of the hypoxia treated cardiac muscle specimen was made, activity subbands originated regularly in the isoyme patterns of LDH, with the regularity of LDH1 (0 subband), LDH2 (0-1 subbands), LDH3 (0-2 subbands), LDH4 (1-3 subbands), LDH5 (2-4 subbands). After adding appropriate amount of NAD+ to the hypoxia treated cardiac muscle specimen, PAGE showed the subbands of four isoymes (LDH2-LDH5) reduced or even totally disappeared in the isozyme patterns.
CONCLUSIONSThe negative feedback regulation of coenzymization and decoenzymization of LDH isozymes is one of the mouse stress responses to slight hypoxia.
Animals ; Hindlimb ; enzymology ; Hypoxia ; enzymology ; Isoenzymes ; metabolism ; L-Lactate Dehydrogenase ; classification ; metabolism ; Liver ; enzymology ; Mice ; Muscles ; enzymology ; Myocardium ; enzymology ; Random Allocation
2.Glycolytic and fatty acid metabolic enzyme changes early after acute myocardial ischemia.
Jing GONG ; Li-huan LI ; Wei-dong PEI ; Hong-yue WANG ; Ying-li ZHENG ; Gui-ying ZHOU ; Shi-yong SHI ; Bin GUAN ; Yan-wan ZHANG ; Zuo-xiang HE
Chinese Journal of Cardiology 2006;34(6):546-550
OBJECTIVETo explore the changes of mRNA and protein expressions of glycolytic and fatty acid metabolic enzymes early after acute myocardial ischemia.
METHODSTwelve dogs were randomly divided into 3 groups (sham, 20 min ischemia and 40 min ischemia, n = 4 each). Myocardial samples from ischemic and nonischemic zone were obtained for histology examination, and the mRNA expressions for Phosphofructokinase (PFK), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), GLUT1, GLUT4, Medium-chain acyl-CoA dehydrogenase (MCAD) and Heart-fatty acid binding protein (H-FABP) were determined by Real Time PCR-SYBR Green RT-PCR. GLUT1 protein expression was determined by immunohistochemistry. The apoptotic cardiomyocytes was evaluated by TUNEL.
RESULTSCompared to sham hearts, H-FABP mRNA was decreased in nonischemic and ischemic zone (P < 0.05) while GLUT1 mRNA expression was significantly increased in nonischemic and ischemic zone (P < 0.05) in dogs underwent 20 and 40 min ischemia. PFK mRNA tended to be higher in ischemic myocardium (P = 0.065) and GAPDH, MCAD as well as GLUT4 remained unchanged post ischemia (all P > 0.05). Positive GLUT1 protein staining was visualized in ischemic myocardium of hearts underwent 20 and 40 min ischemia. The myocardial apoptosis cells was 6.4% +/- 0.9% in sham hearts, 28.0% +/- 3.7% in hearts underwent 20 min ischemia (P < 0.05 vs. sham) and 38.4% +/- 1.9% in hearts underwent 40 min ischemia (P < 0.05 vs. sham).
CONCLUSIONSSignificant down and up-regulated glycolytic and fatty acid metabolic enzymes early after myocardial ischemia suggested that these enzymes might play an important role in acute myocardial ischemia.
Animals ; Disease Models, Animal ; Dogs ; Fatty Acids ; metabolism ; Glycolysis ; Myocardial Ischemia ; enzymology ; Myocardium ; enzymology ; RNA, Messenger ; genetics
3.Mitochondrial aldehyde dehydrogenase in myocardial ischemia-reperfusion injury: from bench to bedside.
Jiao-Jiao PANG ; Linzi A BARTON ; Yu-Guo CHEN ; Jun REN
Acta Physiologica Sinica 2015;67(6):535-544
Acute myocardial infarction is one of the major causes of mortality worldwide. Reperfusion in a timely fashion is the most effective way to limit infarct size. However, reperfusion can itself prompt further myocardial injury. This phenomenon is commonly known as myocardial ischemia-reperfusion (IR) injury. Mitochondrial aldehyde dehydrogenase (ALDH2) is an enzyme metabolizing acetaldehyde and toxic aldehydes. Increasing evidence has revealed a cardioprotective role of ALDH2 in myocardial IR injury. Evidence from animal studies has shown that ALDH2 diminishes acute myocardial infarct size, ameliorates cardiac dysfunction and prevents reperfusion arrhythmias. The activity of ALDH2 is severely compromised if it is encoded by the mutant ALDH2*2 gene, with an incidence of approximately 40% in Asian populations. Epidemiological surveys in the Asian population have depicted that ALDH2 polymorphism is closely associated with higher prevalence of acute myocardial infarction and coronary artery disease. Therefore, targeting ALDH2 may represent a promising avenue to protect against IR injury. This review recapitulates the underlying mechanisms involved in the protective effect of ALDH2 in cardiac IR injury. Translational potential of ALDH2 in the management of coronary heart disease is also discussed.
Aldehyde Dehydrogenase
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metabolism
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Animals
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Heart
;
physiopathology
;
Humans
;
Mitochondria, Heart
;
enzymology
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Myocardial Reperfusion Injury
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Myocardium
;
pathology
4.The risk prediction value of paraquat poisoning dose, urine protein and myocardial enzymes.
Yi Wei SU ; Yi Min LIU ; Jin Wei ZHANG ; Li Ping ZHOU ; Wei Jia DU ; Zhi WANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2023;41(1):8-13
Objective: To explore the value of paraquat (PQ) intake, urine protein and myocardial enzyme indexes in judging the prognosis of patients with acute PQ poisoning. Methods: From September to December 2021, all 201 patients with acute PQ poisoning admitted to Guangzhou Twelfth People's Hospital from January 2010 to December 2019 were selected as the research objects. Based on follow-up results 60 days after poisoning, the research objects were divided into survival group (n=78) and death group (n=123) . The differences in information about poisoning, treatment plan, PQ intake, urine protein, creatine kinase, creatine kinase isoenzyme, lactate dehydrogenase, and α-hydroxybutyrate dehydrogenase between the two groups of patients were compared and analyzed. Logistic regression and Cox regression were used to analyze the correlation between poisoning outcome and PQ intake, urine protein and myocardial enzymes. ROC curve and principal component analysis were used to explore high-efficiency indicators for predicting the outcome of acute PQ poisoning. Results: The PQ intake[50 (20, 100) ml], urine protein (total rank 15570.50) , creatine kinase[ (336.36±261.96) U/L], creatine kinase isoenzyme[ (43.91±43.74) U/L], lactate dehydrogenase [ (346.01±196.50) U/L], α-hydroxybutyrate dehydrogenase content[ (271.23±11.92) U/L] of patients in the death group were all higher than the survival group[15 (10, 20) ml, 4730.50, (187.78±178.06) U/L, (18.88±15.50) U/L, (190.92±60.50) U/L, (152.60±48.34) U/L, respectively] (P<0.05) . The outcome of acute PQ poisoning was positively correlated with PQ intake, urine protein, creatine kinase, creatine kinase isoenzyme, lactate dehydrogenase, and α-hydroxybutyrate dehydrogenase (P<0.05) . Multivariate logistic regression and multivariate Cox regression analysis showed that creatine kinase, creatine kinase isoenzyme, lactate dehydrogenase and α-hydroxybutyrate dehydrogenase was positively correlated with the prognosis of patients with acute PQ poisoning (P<0.05) . ROC curve analysis and principal component analysis showed that the combined indexes of PQ intake, urine protein and myocardial enzymes had the highest efficacy and weight in judging the prognosis of patients (AUC=0.91, weight coefficient=0.19, sensitivity=0.76, specificity=0.89) . When the combined score was ≥4, the probability of accurately predicting the death of patients was as high as 91% (positive predictive value=0.91) . Conclusion: PQ intake, urine protein combined with creatine kinase, creatine kinase isoenzyme, lactate dehydrogenase, and α-hydroxybutyrate dehydrogenase has high value in predicting the prognosis of patients with acute PQ poisoning.
Humans
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Creatine
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Creatine Kinase
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Isoenzymes
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Lactate Dehydrogenases
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Paraquat/poisoning*
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Prognosis
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Retrospective Studies
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Myocardium/enzymology*
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Urine/chemistry*
5.Effect of interleukin-2 on the activity of Ca2+ ATPase and Na+/K+ ATPase of sarcoplasmic reticulum and sarcolemma.
Chun-Mei CAO ; Qiang XIA ; Chen FU ; Hui-Di JIANG ; Zhi-Guo YE ; Yue-Liang SHAN ; Jun-Zhu CHAN
Acta Physiologica Sinica 2003;55(1):83-90
The purpose of the present study was to investigate whether interleukin-2 (IL-2) changes the activity of sarcoplasmic reticulum (SR) Ca(2+) ATPase, sarcolemmal Ca(2+)ATPase and Na(+)/K(+) ATPase by measuring the Pi liberated from ATP hydrolysis with colorimetrical methods. It was shown that the activity of Ca(2+)ATPase in SR from IL-2-perfused (10, 40, 200, 800 U/ml) rat heart increased dose-dependently. After incubation of the SR with ATP (0.1 approximately 4 mmol/L), the activity of SR Ca(2+)ATPase increased dose-dependently in the control group. In the SR from 200 U/ml IL-2-perfused hearts, the activity of Ca(2+)ATPase was much higher than that in the control group. On the other hand, incubation of the SR with Ca(2+) (1 approximately 40 micromol/L) increased the activity of SR Ca(2+) ATPase in the control group. The activity of SR Ca(2+)ATPase of IL-2-perfused hearts was inhibited as the function to Ca(2+). Pretreatment with specific kappa-opioid receptor antagonist nor-BNI (10 nmol/L) for 5 min attenuated the effect of IL-2 (200 U/ml) on the activity of SR Ca(2+) ATPase. After pretreatment with pertussis toxin (PTX, 5 mg/L) or U73122 (5 micromol/L), IL-2 failed to increase SR Ca(2+)ATPase activity. The activity of SR Ca(2+)ATPase was not changed by incubation of SR isolated from normal hearts with IL-2. Perfusion of rat heart with IL-2 did not affect the activity of sarcolemmal Ca(2+)ATPase and Na(+)/K(+)ATPase. It is concluded that perfusion of rat heart with IL-2 increases the activity of SR Ca(2+)ATPase dose-dependently, which is mainly mediated by cardiac kappa-opioid receptor pathway including a PTX sensitive Gi-protein and phospholipase C. IL-2 increases the activity of SR Ca(2+)ATPase as the function to ATP, but inhibits the activity of SR Ca(2+)ATPase as the function to Ca(2+). IL-2 has no effect on the activity of sarcolemmal Ca(2+)ATPase and Na(+)/K(+)ATPase.
Animals
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Interleukin-2
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pharmacology
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Male
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Myocardium
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enzymology
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Rats
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Rats, Sprague-Dawley
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Sarcolemma
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enzymology
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Sarcoplasmic Reticulum
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enzymology
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Sarcoplasmic Reticulum Calcium-Transporting ATPases
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metabolism
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Sodium-Potassium-Exchanging ATPase
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metabolism
6.Expression of NOS III mRNA in different tissues of spontaneously hypertensive rats using RNA array.
Nai-yun CHEN ; Shen-jiang HU ; Hai-tao DONG
Journal of Zhejiang University. Medical sciences 2004;33(5):443-448
OBJECTIVETo evaluate the expression of nitric oxide synthase III (NOS III) mRNA in the heart, aorta, kidney and liver of spontaneously hypertensive rats (SHR).
METHODSTwo hundred and ninety-four total RNA samples were obtained from the tissues of ventricle, aortic smooth muscle, kidney and liver of SHR and normotensive rats (Wistar-Kyoto rats, WKY). RNA array was used to determine the mRNA levels of NOS III of the two groups.
RESULTSCompared with WKY, the systolic blood pressure increased significantly in SHR at 6-week-old, 8-week-old, 10-week-old and 12-week-old [(158.50 +/-7.69 vs 108.67 +/-5.89) mmHg, (174.33 +/-4.46 vs 128.50 +/-4.97) mmHg, (198.00 +/-13.45 vs 142.00 +/-3.58) mmHg, (216.67 +/-8.91 vs 141.17 +/-4.92) mmHg, P<0.01], and the ventricle/body weight ratio was significant higher at 10-week-old and 12-week-old [(4.08 +/-0.17 vs 3.59 +/-0.11, 4.05 +/-0.18 vs 3.40 +/-0.19)mg/g, P<0.01]. In the heart tissue and the kidney, the mRNA levels of NOS III were significantly increased at 6-week-old, 8-week-old, 10-week-old and 12-week-old (1.12 +/-0.18 vs 0.90 +/- 0.15, 1.46 +/- 0.34 vs 1.06 +/-0.18, 1.66 +/- 0.31 vs 1.21 +/- 0.30, 1.98 +/- 0.40 vs 1.31 +/-0.38, P <0.05) and at 4-week-old, 6-week-old, 8-week-old, 10-week-old and 12-week-old (1.10 +/- 0.21 vs 0.81 +/-0.11, 1.28 +/-0.18 vs 0.95 +/-0.13,1.31 +/-0.23 vs 0.99 +/-0.23, 1.70 +/-0.30 vs 1.08 +/-0.25, 1.83 +/-0.33 vs 1.15 +/-0.20, P<0.05 or P<0.01), respectively. There was no significant difference of the NOS III expression in the liver and no significant signals were detected in the aortic smooth muscle.
CONCLUSIONThe results provide the evidence of the increased expression of NOS III in different tissues in SHR and suggests the progressive nature of essential hypertension.
Animals ; Hypertension ; enzymology ; genetics ; Kidney ; enzymology ; Liver ; enzymology ; Male ; Myocardium ; enzymology ; Nitric Oxide Synthase ; biosynthesis ; genetics ; Nitric Oxide Synthase Type III ; Oligonucleotide Array Sequence Analysis ; methods ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY
7.Acute lung injury and changes of myocardial ATP enzymes induced by lipopolysaccharide in aging rats.
Jian ZHANG ; Ye-wei DU ; Ren-yu SUN ; Shi-wen WANG
Acta Academiae Medicinae Sinicae 2003;25(3):320-323
OBJECTIVETo investigate whether acute lung injury (ALI) and changes of myocardial ATP enzymes were induced by intravenous or intraventricle of left heart injection of lipopolysaccharide (LPS) in aging rats.
METHODS40 male Wistar rats were used for reproducing aging animal model. Aging rats were randomly divided into aging control group (n = 8), ALI group (LPS, 5 mg/kg body weight intravenous injection, n = 16), and LPS group (same dosage LPS, intraventricle of left heart injection, n = 16). The samples (blood, lung and heart) were collected at 2, 6 hours after LPS or saline administration.
RESULTSCompared with aging control, protein content in bronchial alveolar lavage fluid (BALF), ratio of lung wet/dry weight and the LA, NO2-/NO3- and MDA contents in blood were increased markedly (P < 0.01) at 2, 6 hours in ALI group. The GSH-Px, Na(+)-K(+)-ATPase activities in lung tissue were decreased significantly (P < 0.01), but NO2-/NO3- content in lung tissue was increased obviously (P < 0.01) at 2 hours in ALI group. These changes were maintained until at 6 hours after LPS administration. The above parameters were no obviously changes in myocardium at 2 hours after LPS administration in ALI group. But at 6 hours, MDA content was increased obviously (P < 0.01); Na(+)-K(+)-ATPase, Ca(2+)-Mg(2+)-ATPase and GSH-Px activities in myocardium were decreased markedly (P < 0.01). While in LPS group, only NO2-/NO3- contents were increased (P < 0.05) in blood and lung tissue as well as Na(+)-K(+)-ATPase activity in lung tissue were decreased (P < 0.05), another parameters had no obvious changes.
CONCLUSIONSALI was obviously formed by intravenous injection LPS after 2, 6 hours in aging rats. Myocardial enzyme etc decreased only at 6 hours in ALI group. But above parameters were no obviously changes in LPS group. It was suggested that there was probable myocardial damage in rats of ALI group, and it was mainly induced by ALI.
Aging ; Animals ; Glutathione Peroxidase ; metabolism ; Lipopolysaccharides ; Male ; Myocardium ; enzymology ; Rats ; Rats, Wistar ; Respiratory Distress Syndrome, Adult ; chemically induced ; enzymology ; Sodium-Potassium-Exchanging ATPase ; metabolism
8.Determination of myocardial enzymes to assess acute CO poisoning-induced myocardial damage.
Chinese Journal of Industrial Hygiene and Occupational Diseases 2004;22(6):469-470
Adolescent
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Adult
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Aged
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Aged, 80 and over
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Carbon Monoxide Poisoning
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enzymology
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Case-Control Studies
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Female
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Humans
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Male
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Middle Aged
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Myocardium
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enzymology
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Young Adult
10.Analysis of the inhibitory effect of gypenoside on Na(+), K (+)-ATPase in rats' heart and brain and its kinetics.
Xiao-yan HAN ; Hong-bo WEI ; Fu-cheng ZHANG
Chinese journal of integrative medicine 2007;13(2):128-131
OBJECTIVETo study the effects of gypenoside (Gyp) on the activity of microsomal Na(+), K(+)-ATPase in rat's heart and brain in vitro.
METHODSThe microsomal Na(+), K(+)-ATPase was prepared from rat's heart and brain by differential centrifugation. The activity of microsomal Na(+), K(+)-ATPase was assayed by colorimetric technique. Enzyme kinetic analysis method was used to analyze the effect of Gyp on the microsomal Na(+), K(+)-ATPase of rats.
RESULTSGyp reversibly inhibited the brain and heart's microsomal Na(+), K(+)-ATPase in a concentration-dependent manner, and showed a more potent effect on enzyme in the brain. The IC(50) of Gyp for the heart and brain were 58.79+/-8.05 mg/L and 52.07+/-6.25 mg/L, respectively. The inhibition was enhanced by lowering the Na(+), or K(+) concentrations or increasing the ATP concentration. Enzyme kinetic studies indicated that the inhibitory effect of Gyp on the enzyme is like that of competitive antagonist of Na(+), the counter-competitive inhibitor for the substrate ATP, and the mixed-type inhibitor for K(+).
CONCLUSIONGyp displays its cardiotonic and central inhibitory effects by way of inhibiting heart and brain's microsomal Na(+), K(+)-ATPase activities in rats.
Adenosine Triphosphate ; pharmacology ; Animals ; Brain ; enzymology ; Enzyme Inhibitors ; pharmacology ; Gynostemma ; Kinetics ; Male ; Myocardium ; enzymology ; Plant Extracts ; pharmacology ; Rats ; Rats, Wistar ; Sodium-Potassium-Exchanging ATPase ; antagonists & inhibitors