Background: This experimental study examined the effect of the antioxidant drug â€œU-74389Gâ€, on a rat model and particularly in a liver ischemia - reperfusion protocol. The effects of that molecule were studied biochemically using blood mean creatine kinase - MB (CK-MB) levels.
Methods: 40 rats of mean weight 231.875 g were used in the study. CK-MB levels were measured at 60 min of reperfusion (groups A and C) and at 120 min of reperfusion (groups B and D). The drug U-74389G was administered only in groups C and D.
Results: U-74389G administration kept significantly increased the CK-MB levels by 31.60% + 13.10% (p = 0.0148). Reperfusion time kept non-significantly increased the CK-MB levels by 5.40% + 14.09% (p= 0.6358). However, U-74389G administration and reperfusion time together kept non-significantly increased the CK-MB levels by 14.50% + 8.16% (p=0.0745).
Conclusions: U-74389G administration reduced at non-significantly increased the CK-MB levels, getting on decline them from significant to non-significant level

Panoulis, Constantinos

Toutouzas, Konstantinos

Triantafyllou, Aggeliki

Tsompos, Constantinos

Zografos, George

ASD Publisher (E-Journals)

Research Journal of Pharmacology and Toxicology 02[02] 2016 www.asdpub.com/index.php/rjpt                                                            ISSN-XXXX-XXXX (Online)                                                  © ASD Publisher All rights reserved.                                                                                                                                                                                                                         4 Original Article  The effect of the antioxidant drug “U-74389G” on creatine kinase - MB levels during ischemia reperfusion injury in rats   C. Τsompos*1, C. Panoulis2, K Τοutouzas3, A. Triantafyllou4, G. Ζografos5  and A. Papalois6  1Consultant A, Department of Obstetrics & Gynecology, Mesologi County Hospital, Etoloakarnania, Greece 2Assistant Professor, Department of Obstetrics & Gynecology, Aretaieion Hospital, Athens University, Attiki, Greece 3Assistant Professor, Department of Surgery, Ippokrateion General Hospital, Athens University, Attiki, Greece 4Associate Professor, Department of Biologic Chemistry, Athens University, Attiki, Greece 5Professor, Department of Surgery, Ippokrateion General Hospital, Athens University, Attiki, Greece  6Director, Experimental Research Centre ELPEN Pharmaceuticals, S.A. Inc., Co., Pikermi, attiki, Greece   *Corresponding Author Tsompos Constantinos  Department of Obstetrics & Gynecology, Mesologi County Hospital, Etoloakarnania,  Greece E-mail: Tsomposconstantinos@gmail.com   Keywords: Ischemia,  U-74389G,  Creatine kinase - MB,  Reperfusion     1. Introduction The chemical group of lazaroid is a C21-amino-steroid complex, devoid of effect on mineralocorticoid and carbohydrate metabolism (glucoactive activity)[1]. Contrary, this complex has a powerful effect against the pathologic lipoperoxidation occurred on lipid membranes. This effect is performed by a steroid-like mechanism but devoid of the side effects typical of high-dose steroids as methylprednisolone[2]. All the lazaroid members act as "scavengers" of oxygen free radicals (ROS) such as hydroxyl radical, lipid peroxides and superoxide anion. Along, it inhibits the lipoxygenase and the production and release of arachidonic acid. The U-74389G is the most famous antioxidant agent of that family. The U-74389G implicates over 254 published biomedical studies at present. The methodology at least of the 46 (18.11%) of these experiments are classified at the tissue ischemia-reperfusion (IR) style. The assumption concerned whether the U-74389G can reverse induced IR injuries in tissues, surrounding organs or even the patients' health. Common affairs were the drug reaction rapidity, the timing of its administration and the dosage height. This antioxidant agent may be proved more beneficial than described. So specific matters are always hardly met in bibliography. A meta-analysis of 35 published related studies yielded a certain numeric efficacy of U-74389G (Table 1). This certain biomedical work tested the effect of U-74389G on a rat liver model. The U-74389G effect was calculated on mean creatine kinase - MB (CK-MB) levels.    2. Materials and methods 2.1 Animal preparation Legal vet licenses were ascribed under the 3693/November 12, 2010 & 14/January 10, 2012 decisions by the local Prefecture in which the acknowledged Co. Inc SA belongs. All the consumed and non substances, settings and equipment were offered by that Co. at Pikermi - Attiki. The Albino female Wistar rats were managed by the precicted humanistic care. Ad libitum diet accompanied the 7 days pre-experimental normal housing in laboratory. Prenarcosis engaged the nonstop intra-experimental care including general anesthesia[3], oxygen supply, electrocardiogram and acidometry. Post-experimental euthanasia excluded awakening and preservation of the animals. The rats were successively delivered to four experimental groups; each one containing 10 animals. Thus, the following protocol of IR was performed: The ischemia stage of 45 min preceded and was common at all groups. However, it was followed by reperfusion for: 60 min for group A; 120 min for group B; 60 min with immediate U-74389G intravenous (IV) administration for group C and 120 min with immediate U-74389G IV administration for group D. The dose of U-74389G was assessed at 10 mg/Kg body mass of animals. Ischemia was induced by laparotomic clamping inferior aorta over the renal arteries with forceps for 45 min. The clamp removal restored the inferior aorta patency and reperfusion. The U-74389G was administered just at initiation of reperfusion; through inferior vena cava catheter. The CK-MB levels were assessed at 60th min of reperfusion for A,C groups and at 120th min of reperfusion for B,D groups. The fourty female Wistar albino rats were described with mean weight (W): 231.875 g [Standard Deviation (SD): 36.59703 g], with minimum W: 165 g and maximum W: 320 g. Rats' W could be perhaps a confusing factor, e.g. the more obese rats, the higher CK-MB levels. This assumption was also tested. 2.2 Control groups The known preceded ischemia of 45 min for the 20 control rats (mW: 252.5 g [SD: 39.31988 g]) was followed by reperfusion.  Group A: Reperfusion lasting 60 min (10 controls rats) concerned mW: 243 g [SD: 45.77724 g] and mean CK-MB levels: 299.4 UI/L [SD: 176.6592 UI/L] (Table 2).         Abstract Background: This experimental study examined the effect of the antioxidant drug “U-74389G”, on a rat model and particularly in a liver ischemia - reperfusion protocol. The effects of that molecule were studied biochemically using blood mean creatine kinase - MB (CK-MB) levels.  Methods: 40 rats of mean weight 231.875 g were used in the study. CK-MB levels were measured at 60 min of reperfusion (groups A and C) and at 120 min of reperfusion (groups B and D). The drug U-74389G was administered only in groups C and D.  Results: U-74389G administration kept significantly increased the CK-MB levels by 31.60% + 13.10% (p = 0.0148). Reperfusion time kept non-significantly increased the CK-MB levels by 5.40% + 14.09% (p= 0.6358). However, U-74389G administration and reperfusion time together kept non-significantly increased the CK-MB levels by 14.50% + 8.16% (p=0.0745). Conclusions: U-74389G administration reduced at non-significantly increased the CK-MB levels, getting on decline them from significant to non-significant level.    C. Τsompos et al / The effect of the antioxidant drug “U-74389G” on creatine kinase - MB levels during ischemia reperfusion injury in rats  © ASD Publisher All rights reserved.                                                                                                                                                                                                                         5 Group B: Reperfusion lasting 120 min (10 controls rats) concerned mW: 262 g [SD: 31.10913 g] and mean CK-MB levels: 315 UI/L [SD: 189.6535 UI/L] (Table 2).  Lazaroid (L) group The known preceded ischemia of 45 min for the 20 L rats (mW: 211.25 g [SD: 17.53755 g] was followed by reperfusion in the beginning of which 10 mg U-74389G /kg body W were IV administered.  Group C: Reperfusion lasting 60 min (10 L rats) concerned mW: 212.5 g [SD: 17.83411 g] and mean CK-MB levels: 451.3 UI/L [SD: 54.75207 UI/L] (Table 2).         Group D: Reperfusion lasting 120 min (10 L rats) concerned mW: 210 g [SD: 18.10463 g] and mean CK-MB levels: 395.9 UI/L [SD: 141.9667 UI/L] (Table 2). 2.3 Statistical analysis Every weight and CK-MB level group was compared with each other by statistical standard t-tests (Table 3). Any significant difference among CK-MB levels, was investigated whether owed in any potent significant weight one. The application of generalized linear models (glm) with dependant variable the CK-MB levels was followed. The 3 independent variables were the U-74389G or no drug administration, the reperfusion time and both variables in combination. Inserting the rats weight also as an independent variable at glm analysis, a non significant relation resulted in (p=0.0723), so as to further investigation was not needed.   3. Results The glm resulted in: U-74389G administration kept significantly increased the CK-MB levels by 116.4 UI/L [210.9841 UI/L - 21.81587 UI/L] (p= 0.0172). This finding was in accordance with standard t-test (p=0.0124). Reperfusion time kept non-significantly increased the CK-MB levels by 19.9 UI/L [-121.707 UI/L - 81.90697 UI/L] (P= 0.6945), also in accordance with the results of standard t-test (p=0.5772). However, U-74389G administration and reperfusion time together kept non-significantly increased the CK-MB levels by 53.41818 UI/L [-5.545847 UI/L - 112.3822 UI/L] (p= 0.0745). Reviewing the above and table 3, the tables 4 and 5 sum up concerning the alteration influence of U-74389G versus reperfusion time.  Table 1: The U-74389G influence (+SD) on the levels of some seric variables [3] concerning reperfusion (rep) time Variable 1h rep p-value 1.5h rep p-value 2h rep p-value interaction of U-74389G and rep p-value WBCC +22.99%+12.45% 0.0914 +30.85%+11.14% 0.0045 +38.70%+17.39% 0.0185 +23.45%+6.28% 0.0004 RBCC   +1.39%+0.71% 0.7161 +0.64%+0.32% 0.8106 -0.10%+0.05% 0.9762 +1.05%+0.53% 0.4911 Hematocrit +5.58%+3% 0.0852 +4.73%+2.25% 0.0435 +3.89%+3.44% 0.2608 +3.16%+1.33% 0.0196 Hemoglobin +5.2%+2.8% 0.0925 +3.9%+2.1% 0.0604 +2.7%+3.2% 0.3544 +2.5%+1.3% 0.0423 MCH +1.77%+0.96% 0.0663 +2.40%+0.57% 0.0001 +3.03%+0.71% 0.0003 1.33%+0.36% 0.0005 MCV +2.12%+1.16% 0.0663 +2.88%+0.69% 0.0001 +3.64%+0.85% 0.0003 +1.6%+0.43% 0.0005 MCHC -0.5%+0.74% 0.4820 -0.95%+0.63% 0.1124 -1.4%+1.12% 0.1603 -0.69%+0.37% 0.0655 RbcDW -6.13%+3.73% 0.0667 -4.96%+2.27% 0.0175 -3.80%+3.07% 0.1383 -2.54%+1.39% 0.679 Platelet count -17.79%+9.40% 0.0647 -12.83%+5.79% 0.0303 -7.88%+7.83% 0.2939 -6.12%+3.58% 0.0857 MPV +9.29%+5.07% 0.0663 +12.77%+3.07% 0.0001 +16.25%+3.81% 0.0003 +7.09%+1.91% 0.0005 Platelet-crit +3.80%+9.87% 0.6373 +9.23%+6.29% 0.1064 +14.66%+9.03% 0.0833 +6.72%+3.73% 0.0712 PDW +1.1%+0.88% 0.2368 +1.79%+0.76% 0.0314 +2.49%+1.33% 0.0807 +0.96%+0.46% 0.0396 Glucose -6.41%+3.50% 0.0663 -8.57%+2.06% 0.0001 -10.74%+2.52% 0.0003 -4.76%+1.28% 0.0005 Urea -8.51%+4.64% 0.0663 -11.35%+2.73% 0.0001 -14.19%+3.32% 0.0003 -6.31%+1.70% 0.0005 Creatinine -15.96%+8.71% 0.0663 -21.02%+5.06% 0.0001 -26.09%+6.12% 0.0003 -11.69%+3.16% 0.0005 Uric acid +20.86%+14.44% 0.1614 +15.43%+9.10% 0.0960 +10%+12.11% 0.3946 +4.78%+5.64% 0.3873 Total protein                -5.48%+2.99% 0.0663 -7.34%+1.76% 0.0000 -9.20%+2.16% 0.0000 -4.08%+1.10% 0.0000 Albumins -2.70%+1.47% 0.0663 -3.63%+0.87% 0.0001 -4.57%+1.07% 0.0003 -2.02%+0.54% 0.0005 ALT3 +11.25%+6.19% 0.0686 -8.83%+11.25% 0.4168 -28.92%+24.55% 0.2206 -4.98%+6.77% 0.4527 AST +33.94%+15.10% 0.0328 +24.97%+14.34% 0.0593 +16%+22.72% 0.4077 +16.23%+8.58% 0.0583 γGT +19.35%+18.58% 0.2362 +6.82%+14.89% 0.6442 -5.71%+20.10% 0.7809 +1.23%+9% 0.8877 ALP  +22.66%+12.37% 0.0663 +31.91%+7.69% 0.0001 +41.16%+9.65% 0.0003 +17.75%+4.79% 0.0005 ACP -112.54%+20.95% 0.0006 -128.45%+14.84% 0.0000 -144.36%+21.62% 0.0000 -74.45%+9.63% 0.0000 CPK +54.32%+13.75% 0.0012 +35.34%+17.20% 0.0260 +16.37%+30.24% 0.4951 +18.52%+9.44% 0.0770 CK-MB +40.46%+ 16.70% 0.0147 +31.60%+13.10% 0.0148 +22.75%+ 22.59% 0.2865 +14.50%+8.16 % 0.0745 LDH +13.56%+7.40% 0.0663 +18.78%+4.52% 0.0001 +24.01%+5.63% 0.0003 +10.43%+2.82% 0.0005 Sodium   +1.22%+0.66% 0.0707 +0.17%+0.61% 0.7714 -0.87%+1.03% 0.3995 -0.32%+0.36% 0.3693 Potassium -10.12%+4.8% 0.0579 -2.14%+5.06% 0.6730 +5.83%+6.79% 0.3801 +2.07%+3.03% 0.4853 Chloride   -0.58%+0.77% 0.4533 -0.97%+0.53% 0.0879 -1.36%+0.76% 0.1113 -0.75%+0.38% 0.0159 Calcium    0%+1.75% 1 -0.14%+1.10% 0.8782 -0.28%+1.54% 0.8492 +0.14%+0.64% 0.8245 Phosphorus   -2.23%+5.51% 0.7966 -1.61%+3.32% 0.5789 -1%+4.48% 0.8129 -1.09%+2% 0.5771 Magnesium +1.33%+3.59% 0.7033 -0.28%+2.75% 0.9171 -1.90%+5.28% 0.7161 +0.36%+4.58% 0.8228 Amylase -6.28%+3.43% 0.0663 -8.40%+2.02% 0.0001 -10.53%+2.47% 0.0003 -4.67%+1.26% 0.0005 Progesterone -46.14%+25.19% 0.0663 -58.04%+13.99% 0.0001 -69.94%+16.41% 0.0003 -32.29%+8.73% 0.0005 Testosterone +40.35%+28.70% 0.1261 +52.17%+28.69% 0.0451 +64%+44.32% 0.1380 +11.18%+17.97% 0.5245 Mean 2.03%+27.26% 0.2168 0.19%+29.41% 0.1836 -1.63%+33.15% 0.2389 -0.33%+16.23% 0.2016  Table 2: Weight and CK-MB levels and Std. Dev. of groups Groups   Variable Mean Std. Dev    Α Weight 243 g 45.77724 g  CK-MB 299.4 UI/L 176.6592 UI/L B Weight 262 g 31.10913 g  CK-MB 315 UI/L 189.6535 UI/L C Weight 212,5 g 17.83411 g  CK-MB 451.3 UI/L 54.75207 UI/L D Weight 210 g 18.10463 g  CK-MB 141.9667 395.9 UI/L UI/L  C. Τsompos et al / The effect of the antioxidant drug “U-74389G” on creatine kinase - MB levels during ischemia reperfusion injury in rats  © ASD Publisher All rights reserved.                                                                                                                                                                                                                         6 Table 3: Statistical significance of mean values difference for groups (DG) after statistical standard t test application DG Variable Difference p-value A-B Weight -19 g 0.2423  CK-MB -15.6UI/L 0.7558 Α -C Weight 30.5 g 0.0674  CK-MB -151.9 UI/L 0.0113 Α -D Weight 33 g 0.0503  CK-MB -96.5 UI/L 0.2064 Β -C Weight 49.5 g 0.0019  CK-MB -136.3 UI/L 0.0473 Β -P Weight 52 g 0.0004  CK-MB -80.9 UI/L 0.2786 C-D Weight 2.5 g 0.7043 CK-MB 55.4 UI/L 0.3009  Table 4: The restore influence of U-74389G versus reperfusion time    p-values Increase 95% c. in. Reperfusion time t-test glm 151.9 UI/L 29.02519 UI/L - 274.7748 UI/L 1h 0.0182 0.0113 116.4 UI/L 21.81587 UI/L - 210.9841 UI/L 1.5h 0.0124 0.0172 80.9 UI/L -76.49116 UI/L - 238.2912 UI/L 2h 0.2945 0.2786 -19.9 UI/L -121.707 UI/L - 81.90697 UI/L reperfusion temps 0.6945 0.5772 -5.545847 53.41818 UI/L UI/L - 112.3822 UI/L interaction - 0.0745   Table 5: The (%) restore influence of U-74389G in connection with reperfusion time. Increase +SD Reperfusion time p-values 40.46 % + 16.70 % 1h 0.0147 31.60 % +13.10 % 1.5h 0.0148 22.75 % + 22.59 % 2h 0.2865 -5.40% + 14.09% reperfusion temps 0.6358 14.50% + 8.16 % interaction 0.0745  4. Discussion The CK-MB levels can be influenced by ischemic cases. Gonçalves ES et al[4] noted a significant increase of the CK-MB plasma levels after 30 minutes IR in pilot rats treated by ornithin α-ketoglutarate. Jebeli M et al[5] found significantly elevated the CK-MB serum levels in ischemia or myocardial infarction among patients presenting either deterioration of the left ventricular function [(FEVG) < 35%] or acute phase of myocardial infarction (MI), suffered from ventricular tachycardia (VT). Sala MF et al[6] estimated more frequently by 3.95-fold the peak of CK-MB levels > 300 IU/L in acute phase of MI patients suffering from VT. Serruys PW et al[7] noted no case of raised CK-MB levels 10 times over the upper normal limit in patients after the success of their first percutaneous coronary intervention (PCCI) randomly receiving treatment with either fluvastatine 80 mg/kg or placebo one, during 3 to 4 years after hospital exit. Savchuk VI et al[8] calculated increased the total activity of CK-MB by 57.3+11.7 mE/ml (p < 0.001) in coronary blood after myocardial ischemia induction via short run in dogs. Nediani C et al attenuated all the modifications, particularly, the neutrophiles infiltration by aminosteroid administration (4 mg/kg) in coronary IR of pigs. The 21-lazaroid aminosteroids[9] trap free radicals and work as membrane stabilizers. The energy load reduction was reversed in the treated group. Myocardial concentration of malondialdehyde was raised in all animals after the reperfusion, but this effect was significantly marked with aminosteroids treatment. Moreover, an increase in infarctions of ascorbic acid contents and a potency to reduce the serum peroxidation rate exposed in animals treated than untreated ones, indicate an improvement of the antioxidant protection induced by administration of aminosteroids. In addition, the serum rates of CK-MB isoenzyme suggest the possibility of the aminosteroids to attenuate the modifications of the membrane permeability induced by IR damage. The aminosteroid treatment is effective in the reduction of morphological and biochemical deteriorations which occur in an IR myocardium.  4. Conclusion U-74389G administration reduced at non-significantly increased the CK-MB levels, getting on decline them from significant to non-significant level.    Acknowledgment This study was funded by Scholarship by the Experimental Research Centre ELPEN Pharmaceuticals (E.R.C.E), Athens, Greece. The research facilities for this project were provided by the aforementioned institution.  References [1] https://www.caymanchem.com/app/template/Product.vm/catalog/75860.  [2] Fenglin Shi, Jennifer Cavitt, Kenneth L Audus. 21-aminosteroid and 2-(aminomethyl)chromans inhibition of arachidonic acid-induced lipid peroxidation and permeability enhancement in bovine brain microvessel endothelial cell monolayers. Free Radical Biology and Medicine, 1995; 19(3): 349-357. [3] Tsompos C, Panoulis C, Toutouzas K, Triantafyllou A, Zografos G, Papalois A. The Antioxidant Drug “U-74389g” Effect on Alanine Aminotransferase Levels. J Anal Pharm Res 2016; 4(2): 00095.  [4] Gonçalves Rabelo ES, CM, Prado Neto AX, et al: effet de court-terme ornithine alpha-cétoglutarate prétraitement sur ischémie intestinale -la reperfusion chez les rats. L'Acta Cir Bras. 2011; 26 Suppl 1:2-7. [5] Ghazinoor Jebeli M, M, Mandegar MH, et al: Effet du milrinone sur résultat à court terme des patients avec dysfonction myocardique subissant coronary artery bypass graft: a randomized controlled trial. Cardiol J. 2010; 17(1):73-8. C. Τsompos et al / The effect of the antioxidant drug “U-74389G” on creatine kinase - MB levels during ischemia reperfusion injury in rats  © ASD Publisher All rights reserved.                                                                                                                                                                                                                         7 [6] Sala MF, Bárcena JP, Rota JI, et al: Tachycardie ventriculaire soutenue en tant que marqueur de l'insuffisance de la perfusion myocardique durant la phase aiguë d'infarctus du myocarde. Clin Cardiol. 2002 Jul; 25(7):328-34. [7] Serruys PW, de Feyter Macaya P, C, et al: La fluvastatine pour la prévention des événements cardiaques après réussite première intervention coronaire percutanée : un essai contrôlé randomisé. JAMA. 2002 Jun 26; 287(24):3215-22.                                                      [8] Savchuk VI , Vinogradov AV , Pozin VM , et al : activité du total de la créatine phosphokinase sérique et ses mo fraction dans le débit sanguin coronaire réversible disorders in dogs dans une expérience chronique. Kardiologiia. 1980 Jan; 20(1):58-60. [9] Nediani C , Perna suis , Liguori P , et al : effets bénéfiques de la 21-74389aminosteroid U G sur la lésion d'ischémie-reperfusion dommages dans des coeurs de porcs. J Mol Cell Cardiol. 1997 Oct; 29(10):2825-35.  

The effect of the antioxidant drug â€œU-74389Gâ€ on creatine kinase - MB levels during ischemia reperfusion injury in rats

http://asdpub.com/index.php/rjpt/article/download/350/230/

The effect of the antioxidant drug â€œU-74389Gâ€ on creatine kinase - MB levels during ischemia reperfusion injury in rats

Abstract

Similar works

Full text

Available Versions

ASD Publisher (E-Journals)