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The effect of alpha-lipoic acid (ALA) on early stages of atherosclerosis in rats with diabetes
Abstracts – XIX Congress PTF / Pharmacological Reports 67S (2015) 2–45
Expression of drug metabolizing enzymes along the human gastrointestinal tract Marek Droz´dzik 1, Jette Peters 2, Joanna Lapczuk 1, Marek Ostrowski 3, Sierk Haenisch 4, Werner Siegmund 2, Stefan Oswald 2 1
Department of Experimental and Clinical Pharmacology, Pomeranian Medical University, Szczecin, Poland 2 Department of Clinical Pharmacology, University of Greifswald, Greifswald, Germany 3 Department of General and Transplantation Surgery, Pomeranian Medical University, Szczecin, Poland 4 Institute of Experimental and Clinical Pharmacology, University of Kiel, Kiel, Germany
The oral absorption of many drugs is highly influenced by intestinal biotransformation. To characterize or even predict the impact of intestinal drug processing, the knowledge of intestinal expression levels of enzymes is an essential prerequisite. Therefore, the aim of this study was to comprehensively analyse the expression of clinically relevant phase I and II enzymes, and nuclear receptors along the entire human gastrointestinal tract. Intestinal tissue was collected from 8 organ donors (age: 24–56). In each case 10 samples from the entire gut (duodenum: 1, jejunum: 2, ileum: 3, colon: 4) were taken, and mRNA expression of 12 genes of metabolizing enzymes (CYPs: 8, phase II enzymes: 4) were measured using custom-made TaqMan1 low density arrays. Gene expression was normalized to five house-keeping genes and analyzed using the DDCt-method. Intestinal expression of CYP2C9, CYP2D6, SULT1A, UGT1A and UGT2B7 increased from duodenum to jejunum and was the highest in distal jejunum but markedly lower in ileum and colon. CYP3A4/5 was predominately found in the upper intestine. The intestinal expression rang order of drug metabolizing enzymes was as follows: CYP3A4 > CYP2C9 > CYP2C19 > CYP2D6 = CYP3A5 = CYP2B6, CYP2C8. In all intestinal regions, SULT1A, UGT1A, CYP3A4 and CYP2C9 were the most prominent genes. The intestinal distribution of the expression of SULT1A and UGT1A enzymes and UGT2B7 was as follows: duodenum < jejunum > ileum > colon, whereas for UGT2B15 there was stable but significantly lower expression along GI tract. There is a substantial expression gradient for several enzymes along the human alimentary tract, which may explain regio-selective differences in intestinal drug metabolism. http://dx.doi.org/10.1016/j.pharep.2015.06.064 The effect of alpha-lipoic acid (ALA) on early stages of atherosclerosis in rats with diabetes Marzena Dworacka 1, Hanna Winiarska 1, Anna Wesołowska 1, Saule Iskakova 2, Yergen Kurmambayev 2, Galina Chukanova 2, Bartosz Frycz 3, Paweł P. Jagodzin´ski 3, Grzegorz Dworacki 4 1
Department of Pharmacology, Poznan´ University of Medical Sciences, Poznan´, Poland 2 Department of Pharmacology, Marat Ospanov University of Medical Sciences, Aktobe, Kazakhstan 3 Department of Biochemistry and Molecular Biology, Poznan´ University of Medical Sciences, Poznan´, Poland 4 Department of Clinical Immunology, Poznan´ University of Medical Sciences, Poznan´, Poland Recently the efficacy of alpha-lipoic acid (ALA) on atherosclerosis was suggested, by showing that ALA modulates multiple pathogenic aspects of atherosclerosis. The aim of the study was to
21
examine the effect of ALA on two indicators of early atherosclerosis: mRNA TGF-b expression in tissues and on cEC concentration in peripheral blood of rats with diabetes. The investigation was performed on 28 male Wistar rats with induced diabetes and on 15 non-diabetic animals. ALA or placebo were administered into the stomach once daily within 8 weeks to all of examined animals. TGF-b mRNA expression in aorta and in myocardium was assayed by real-time PCR and cEC count was measured using flow cytometry techniques. It was found that TGF-b mRNA expression in myocardium in ALA-treated rats with diabetes is lower than in placebo-treated rats with diabetes and similar to the control nondiabetic group. TGF-b mRNA expression in aorta of ALA-treated rats with diabetes was higher than in placebo-treated rats with diabetes and in comparison to non-diabetic animals. The percentage of cEC in peripheral blood observed in ALA-treated rats was similar to non-diabetic group and significantly lower than in placebo-treated animals with diabetes. It could be concluded that ALA could be the helpful agent in prevention of diabetesrelated atherosclerosis development. http://dx.doi.org/10.1016/j.pharep.2015.06.065 Effect of choline salicylate DPCM and DPAE in guinea pigs Wojciech Dziewiszek 1, Wojciech Michalski 2, Adam Szela˛g 1, Marek Bochnia 3 1
Department of Pharmacology, Wrocław Medical University, Wrocław, Poland 2 Wrocław University of Technology, Institute of Telecommunications, Teleinformatics and Acoustics, Wrocław, Poland 3 Department of Otolaryngology, Faculty of Dentistry, Wrocław Medical University, Wrocław, Poland The aim of work was an assessment of the effect of choline salicylate i.m. application on the distortion product of cochlear microphonics (DPCM) and acoustic otoemission (DPOAEThe ototoxicity study of choline salicylate (ChS) was performed in 6 guinea pigs. Choline salicylate was administered intramuscularly in the dose 200 mg/kg. CM and DPOAE were received using the new homodyne method before and 60 min after each medicine application. Biopotentials were examined under ketamine/xylazine anesthesia. Measurements were carried out for five fixed intermodulation frequencies f3: 1321, 1875, 2671, 3142 and 5342 Hz. Each of the five frequencies was obtained for 4 different ratios k = f2/f1 = 1.20, 1.25, 1.30, 1.35. The following two tone intensity combinations were used: 50 dB, 60 dB and 55 dB, 65 dB. The measurements were reregistered continuously with the computer system. The effect of the choline salicylate both on the DPCM and DPOAE level and the phase of these signals were investigated. The results show that the DPOAE signal on some frequencies was increased. In addition, by analyzing the phase behavior, was observed the change its course and the reduction of the phase signal stability DPCM and DPOAE. http://dx.doi.org/10.1016/j.pharep.2015.06.066 Differential skeletal effects of trigonelline, an alkaloid present in coffee, in streptozotocin- and streptozotocin/nicotinamide-treated rats Joanna Folwarczna 1, Aleksandra Janas 1, Maria Pytlik 1, Urszula Cegieła 1, Leszek S´liwin´ski 1, Zora Krivosˇı´kova´ 2, Korne´lia Sˇtefı´kova´ 2, Martin Gajdosˇ 2
Expression of drug metabolizing enzymes along the human gastrointestinal tract Marek Droz´dzik 1, Jette Peters 2, Joanna Lapczuk 1, Marek Ostrowski 3, Sierk Haenisch 4, Werner Siegmund 2, Stefan Oswald 2 1
Department of Experimental and Clinical Pharmacology, Pomeranian Medical University, Szczecin, Poland 2 Department of Clinical Pharmacology, University of Greifswald, Greifswald, Germany 3 Department of General and Transplantation Surgery, Pomeranian Medical University, Szczecin, Poland 4 Institute of Experimental and Clinical Pharmacology, University of Kiel, Kiel, Germany
The oral absorption of many drugs is highly influenced by intestinal biotransformation. To characterize or even predict the impact of intestinal drug processing, the knowledge of intestinal expression levels of enzymes is an essential prerequisite. Therefore, the aim of this study was to comprehensively analyse the expression of clinically relevant phase I and II enzymes, and nuclear receptors along the entire human gastrointestinal tract. Intestinal tissue was collected from 8 organ donors (age: 24–56). In each case 10 samples from the entire gut (duodenum: 1, jejunum: 2, ileum: 3, colon: 4) were taken, and mRNA expression of 12 genes of metabolizing enzymes (CYPs: 8, phase II enzymes: 4) were measured using custom-made TaqMan1 low density arrays. Gene expression was normalized to five house-keeping genes and analyzed using the DDCt-method. Intestinal expression of CYP2C9, CYP2D6, SULT1A, UGT1A and UGT2B7 increased from duodenum to jejunum and was the highest in distal jejunum but markedly lower in ileum and colon. CYP3A4/5 was predominately found in the upper intestine. The intestinal expression rang order of drug metabolizing enzymes was as follows: CYP3A4 > CYP2C9 > CYP2C19 > CYP2D6 = CYP3A5 = CYP2B6, CYP2C8. In all intestinal regions, SULT1A, UGT1A, CYP3A4 and CYP2C9 were the most prominent genes. The intestinal distribution of the expression of SULT1A and UGT1A enzymes and UGT2B7 was as follows: duodenum < jejunum > ileum > colon, whereas for UGT2B15 there was stable but significantly lower expression along GI tract. There is a substantial expression gradient for several enzymes along the human alimentary tract, which may explain regio-selective differences in intestinal drug metabolism. http://dx.doi.org/10.1016/j.pharep.2015.06.064 The effect of alpha-lipoic acid (ALA) on early stages of atherosclerosis in rats with diabetes Marzena Dworacka 1, Hanna Winiarska 1, Anna Wesołowska 1, Saule Iskakova 2, Yergen Kurmambayev 2, Galina Chukanova 2, Bartosz Frycz 3, Paweł P. Jagodzin´ski 3, Grzegorz Dworacki 4 1
Department of Pharmacology, Poznan´ University of Medical Sciences, Poznan´, Poland 2 Department of Pharmacology, Marat Ospanov University of Medical Sciences, Aktobe, Kazakhstan 3 Department of Biochemistry and Molecular Biology, Poznan´ University of Medical Sciences, Poznan´, Poland 4 Department of Clinical Immunology, Poznan´ University of Medical Sciences, Poznan´, Poland Recently the efficacy of alpha-lipoic acid (ALA) on atherosclerosis was suggested, by showing that ALA modulates multiple pathogenic aspects of atherosclerosis. The aim of the study was to
21
examine the effect of ALA on two indicators of early atherosclerosis: mRNA TGF-b expression in tissues and on cEC concentration in peripheral blood of rats with diabetes. The investigation was performed on 28 male Wistar rats with induced diabetes and on 15 non-diabetic animals. ALA or placebo were administered into the stomach once daily within 8 weeks to all of examined animals. TGF-b mRNA expression in aorta and in myocardium was assayed by real-time PCR and cEC count was measured using flow cytometry techniques. It was found that TGF-b mRNA expression in myocardium in ALA-treated rats with diabetes is lower than in placebo-treated rats with diabetes and similar to the control nondiabetic group. TGF-b mRNA expression in aorta of ALA-treated rats with diabetes was higher than in placebo-treated rats with diabetes and in comparison to non-diabetic animals. The percentage of cEC in peripheral blood observed in ALA-treated rats was similar to non-diabetic group and significantly lower than in placebo-treated animals with diabetes. It could be concluded that ALA could be the helpful agent in prevention of diabetesrelated atherosclerosis development. http://dx.doi.org/10.1016/j.pharep.2015.06.065 Effect of choline salicylate DPCM and DPAE in guinea pigs Wojciech Dziewiszek 1, Wojciech Michalski 2, Adam Szela˛g 1, Marek Bochnia 3 1
Department of Pharmacology, Wrocław Medical University, Wrocław, Poland 2 Wrocław University of Technology, Institute of Telecommunications, Teleinformatics and Acoustics, Wrocław, Poland 3 Department of Otolaryngology, Faculty of Dentistry, Wrocław Medical University, Wrocław, Poland The aim of work was an assessment of the effect of choline salicylate i.m. application on the distortion product of cochlear microphonics (DPCM) and acoustic otoemission (DPOAEThe ototoxicity study of choline salicylate (ChS) was performed in 6 guinea pigs. Choline salicylate was administered intramuscularly in the dose 200 mg/kg. CM and DPOAE were received using the new homodyne method before and 60 min after each medicine application. Biopotentials were examined under ketamine/xylazine anesthesia. Measurements were carried out for five fixed intermodulation frequencies f3: 1321, 1875, 2671, 3142 and 5342 Hz. Each of the five frequencies was obtained for 4 different ratios k = f2/f1 = 1.20, 1.25, 1.30, 1.35. The following two tone intensity combinations were used: 50 dB, 60 dB and 55 dB, 65 dB. The measurements were reregistered continuously with the computer system. The effect of the choline salicylate both on the DPCM and DPOAE level and the phase of these signals were investigated. The results show that the DPOAE signal on some frequencies was increased. In addition, by analyzing the phase behavior, was observed the change its course and the reduction of the phase signal stability DPCM and DPOAE. http://dx.doi.org/10.1016/j.pharep.2015.06.066 Differential skeletal effects of trigonelline, an alkaloid present in coffee, in streptozotocin- and streptozotocin/nicotinamide-treated rats Joanna Folwarczna 1, Aleksandra Janas 1, Maria Pytlik 1, Urszula Cegieła 1, Leszek S´liwin´ski 1, Zora Krivosˇı´kova´ 2, Korne´lia Sˇtefı´kova´ 2, Martin Gajdosˇ 2