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Study of the effect of glucose on spectroscopy features of doxorubicin anticancer drug
S178
Abstracts
Introduction: The possibility that readily available natural substances have beneficial effect in the prevention and treatment of cancer warrants closer examination. Saffron and its components have been investigated as potential agents for the prevention and treatment of various cancers. These compounds play an important role in regulating cell growth, differentiation, and apoptosis. The aim of this study is to investigate the role of picrocrocin, as a monoterpene aldehayde derived from saffron, on rat mammary tumor induced by N-methyl-N-nitrosourea (NMU). Enkephalin may has a role in the breast cancer cell growth and progression not only in the breast, but also at the neuroendocrine level. Methods: NMU was used for tumor induction on female Wistar rats. Body weight and tumor appearance/size were recorded weekly. Latency period (LP) as the number of days between NMU injection and appearance of the first tumor, tumor incidence (TI) as the percentage of the rats that developed at least one tumor, and mean tumor number per rat (n/t)were determined. In addition EDA specific activity was assayed in the ovary and brain of animals. Results and Conclusion: The results show a decrease in the EDA activity in the ovary and brain of rats after NMU administration. However an increase in EDA activity was observed in the cancerous groups due to the picrocrocin treatment in comparison with control and cancerous groups, without treatment. In addition, tumor size decreased in cancerous group with picrocrocin treatment. In conclusion, our results indicated the beneficial effect of picrocrocin in breast cancer treatment. Keywords: Saffron, Breast cancer, Picrocrocin, Enkephalin degrading enzyme
doi:10.1016/j.clinbiochem.2011.08.784
Poster – [A-10-1096-1] Effect of glycerol, as a chemical chaperone, on hemoglobin glycation by glucose or fructose Asghar Farajzadea, S. Zahra Bathaieb a Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat ModaresIran University, Tehran, Iran b Iran, Tehran, Tarbiat Modares University, Faculty of Medical Sciences, Department of Clinical Biochemistry.P.O.BOX: 14114–111 E-mail addresses: [email protected] (A. Farajzade), [email protected] (S.Z. Bathaie) Introduction: None enzymatic reactions between reducing sugars and proteins produce some complexes, which are named as advanced glycation end products (AGEs). They can play an important role in diabetic complications, cancer, atherosclerosis, aging, prion and Alzheimer diseases, etc. Protocols that result in the decrease of AGEs can help people faced with these diseases. There are some methods to decrease AGEs like controlling carbohydrate`s concentration in the body or using some drugs such as metformin, aspirin and so on. Here, we used chemical chaperones for this purpose. Chemical chaperons are molecules with low molecular weight which are known for the body and deal with proteins to stabilize them against thermal or chemical denaturation. Methods: Glycerol, as a polyol, is selected to prevent hemoglobin glycation by glucose or fructose. Hemoglobin solution, pH 7.4, is incubated separately with glucose or fructose in the presence or absence of glycerol at 37 °C in a shaker incubator. After completion of the reaction, all samples are investigated by different techniques. Results: Incubated Hb with both glucose/fructose and glycerol shows less fluorescence emission in comparison with solution containing glycerol. In addition, there are some changes in the electrophretic
mobility and secondary structure content of hemoglobin in comparison with Hb+ glucose/fructose in the presence or absence of glycerol. Conclusion: AGE formation causes an increase in the fluorescence emission, a decrease in the alpha-helix content and an increase in the electrophoretic mobility of Hb. However, glycerol has preventing effect on these changes and stabilizes hemoglobin against reducing sugars. Keywords: Glycerol, Chemical chaperon, Hemoglobin, AGE, Glycation doi:10.1016/j.clinbiochem.2011.08.785
Poster — [A-10-1101-1] Effects of diabetes on myocardial capillary density and serum biomarkers of angiogenesis in male rats Khazaei Majid, Fallahzadeh Alireza Department of Physiology, Isfahan University of Medical Sciences, Hezar Jarib Ave, Isfahan, Iran E-mail addresses: [email protected] (K. Majid), [email protected] (F. Alireza) Introduction: Cardiovascular disease is one of the main causes of mortality and morbidity in diabetic subjects. This study evaluates the effect of diabetes on myocardial capillary density and some serum angiogenic factors including nitric oxide, vascular endothelial growth factor and its soluble receptors. Methods: Twelve male rats were divided into two groups; control and diabetic (n = 6 each). Diabetes was induced by single dose of streptozotocin (50 mg/kg, intraperitoneal). After 21 days, capillary density in myocardial tissue was evaluated by immunohistochemical staining and reported as capillaries per 2 mm. Blood samples were collected before and after experiment. Results: In diabetic group, serum nitric oxide and soluble vascular endothelial growth factor receptor-2 concentrations were lower and soluble vascular endothelial growth factor receptor-1 was significantly higher than the control group. There was no significant change in serum vascular endothelial growth factor concentration between diabetic and control groups, however, vascular endothelial growth factor/vascular endothelial growth factor receptor-1 ratio was significantly decreased in diabetic animals. Myocardial capillary density in diabetic group was lower than the control group (1549 ± 161 vs. 2156 ± 202/2 mm, respectively). Conclusion: Reduced serum nitric oxide and vascular endothelial growth factor receptor-2 levels, increased serum vascular endothelial growth factor receptor-1 and lower vascular endothelial growth factor/ vascular endothelial growth factor receptor-1 ratio might be responsible for decreased myocardial capillary density in diabetic rats. Keywords: Diabetes, Capillary density, Vascular endothelial growth factor, Nitric oxide, Myocardium doi:10.1016/j.clinbiochem.2011.08.929
Poster — [A-10-1107-1] Study of the effect of glucose on spectroscopy features of doxorubicin anticancer drug Abri Mehraban Fatemeh, Zargar Seyed Jalal, Habibi Rezayee Mehran University of Tehran, Islamic Republic of Iran E-mail addresses: [email protected] (A.M. Fatemeh), [email protected] (Z.S. Jalal), [email protected] (H.R. Mehran) Introduction: Doxorubicin is an anthracycline antibiotic being used in chemotherapy of a wide range of different cancers specifically solid
Abstracts
tumors and leukemia. Glucose is the most common hexose carbohydrate which has specific metabolically roles in biological systems. Method: In this study the interaction of doxorubicin with different glucose concentrations (100, 300 and 500 mg/dl) at room temperature was investigated for 2 h, using ultraviolet spectroscopy technique (200– 600 nm). Results: The results showed that glucose increases the absorbance of doxorubicin at the wavelength of 480 nm and induces hyperchromicity in the absorption spectrum of this drug. Conclusion: This study demonstrated that glucose affects doxorubicin and changes its absorbance at 480 nm, and UV spectra. Since the absorbance at 480 nm corresponds to tetracycline ring of doxorubicin, the results suggested that glucose probably interacts with the tetracycline group of drug molecule.
S179
Keywords: Simvastatin therapy, Prooxidant–antioxidant Balance, Heat shock protein 60, 65, and 70 doi:10.1016/j.clinbiochem.2011.08.931
Poster — [A-10-1141-1] The effect of different concentrations of morphine and extracellular Ca2+ on cell death in PC12 cell line Arezou Rabziaa, Mehri Azadbakhta, Ali Bidmeshki Poura, Hasan Akramia a Department of Biology, Razi University, Kermanshah, Iran E-mail addresses: [email protected] (A. Rabzia), [email protected] (M. Azadbakht)
Keywords: Doxorubicin, Glucose, UV spectroscopy doi:10.1016/j.clinbiochem.2011.08.930
Poster — [A-10-1123-1] Simvastatin therapy reduces prooxidant–antioxidant balance and heat shock protein 60, 65, and 70 antibody titers: Results of a placebo-controlled cross-over trial Barzegar Amini Maral, Mouhebati Mohsen, Ghayour-Mobarhan Majid, Azarpazhooh Mohammad-Reza, Sahebkar Amir-Hosein, Hassanzade-Daloee Mahdi, Momenzade Akram, Tavallaei Shima, Rahsepar Amir-Ali, Khojasteh-Taheri Roshanak Mashhad University of Medical Siences, Mashhad, Iran E-mail addresses: [email protected] (B.A. Maral), [email protected] (M. Mohsen), [email protected] (G.-M. Majid), [email protected] (A. Mohammad-Reza), [email protected] (S. Amir-Hosein), [email protected] (H.-D. Mahdi), [email protected] (M. Akram), [email protected] (T. Shima), [email protected] (R. Amir-Ali), [email protected] (K.-T. Roshanak) Introduction: Oxidative stress and heat shock proteins (HSPs) are thought to play an important role in atherogenesis. The statin group of cholesterol-lowering drugs has been shown to reduce cardiovascular events and possess antioxidant properties and immunomodulatory effects. This study was carried out to evaluate the effects of statin therapy on serum levels of antibodies HSPs and prooxidant– antioxidant balance (PAB), as novel coronary risk factors in dyslipidemic patients. Method: The PAB assay can measure the prooxidant burden and the antioxidant capacity simultaneously in one assay, thereby giving a redox index. We treated 102 dyslipidemic individuals with simvastatin, or a placebo in a double-blind, cross-over, placebo-controlled trial. PAB values and Anti-HSP60, 65, 70, and hs-CRP levels were measured before and after each treatment period. Seventy-seven subjects completed the study. Result: We found that statin therapy was associated with a significant reduction in PAB values (P < 0.001) and serum anti-HSP60, 65, and 70 titers in the dyslipidemic patients (10%, 14%, and 15% decrease, respectively) (p < 0.001). There have been previous reports of reductions in serum CRP with statin treatment, and although median CRP levels were 9% lower on simvastatin treatment, this did not achieve statistical significance. Conclusions: These findings suggest that simvastatin inhibits autoimmune responses and oxidative stress that may contribute to the development of cardiovascular disease.
Introduction: Morphine as a mu opioid drug can induce different effects. Ca2+ as one of the key regulators of cell survival, in response to a wide range of conditions, is capable of inducing cell death. Here, we investigated the effect of different concentrations of morphine together with different concentrations of extracellular Ca2+ on cell death in PC12 cell line. Materials and methods: PC12 cells were cultured in RPMI 1640 culture medium containing 10% fetal bovine serum and different concentrations of morphine (10 E-6 and 10 E-4 M) together with different concentrations of extracellular Ca2+ (0.7, 0.6, 0.5, 0.4, 0.3, 0.2 and 0.00 mM) for 6, 12 and 24 h. PC12 cells that were cultured without morphine were considered as control. Cell viability was assessed by neutral red staining. We used Hoechst/PI nuclear staining and fluorescence microscopy for detection of percentage of apoptotic and necrotic cells. Results: Results indicated that cell viability of PC12 cells in the presence of concentration of 10 E-4 M of morphine together with 0.7 mM of extracellular Ca2+ decreased compared to control (P < 0.05). Hoechst/PI nuclear staining demonstrated that 10 E-4 M of morphine together with 0.7 mM of extracellular Ca2+ were toxic and increased the percentage of apoptotic and necrotic cells. Conclusion: It is showed that high-dose of morphine decreasesd cell viability and enhanced apoptosis and necrosis in PC12 cells, that was dependent on the presence of extracellular Ca2+. Keywords: Morphine, Ca2+, Cell death, Apoptosis, PC12 cell
doi:10.1016/j.clinbiochem.2011.08.932
Poster — [A-10-1147-1] Evaluation of anticoagulant properties of Artemisia dracunculus, Punica granatum and Berberis vulgaris in rat Hosseinzadeh Elyas, Razieh University of Tehran, I.B.B, Laboratory of Genetic Engineering, Islamic Republic of Iran E-mail addresses: [email protected] (H. Elyas), [email protected] (Razieh) Introduction: One of the most important pathways leading to thrombosis is platelet adhesion to extracellular matrix proteins that in turn causes platelet plug formation and eventually formation of blood clot (thrombus) inside a blood vessel that leads to cardiovascular desease. Punica granatum is used as blood dilutor and antidiabetic effect, and Artemisia dracunculus and Berberis vulgaris are used as an anticoaguant and antihyperlipidaemic agents in Iranian folk medicine. In that line, we investigated the inhibitory effects of crude extracts of these plants on the adhesion of the activated platelets to fibrinogen coated plates.
Abstracts
Introduction: The possibility that readily available natural substances have beneficial effect in the prevention and treatment of cancer warrants closer examination. Saffron and its components have been investigated as potential agents for the prevention and treatment of various cancers. These compounds play an important role in regulating cell growth, differentiation, and apoptosis. The aim of this study is to investigate the role of picrocrocin, as a monoterpene aldehayde derived from saffron, on rat mammary tumor induced by N-methyl-N-nitrosourea (NMU). Enkephalin may has a role in the breast cancer cell growth and progression not only in the breast, but also at the neuroendocrine level. Methods: NMU was used for tumor induction on female Wistar rats. Body weight and tumor appearance/size were recorded weekly. Latency period (LP) as the number of days between NMU injection and appearance of the first tumor, tumor incidence (TI) as the percentage of the rats that developed at least one tumor, and mean tumor number per rat (n/t)were determined. In addition EDA specific activity was assayed in the ovary and brain of animals. Results and Conclusion: The results show a decrease in the EDA activity in the ovary and brain of rats after NMU administration. However an increase in EDA activity was observed in the cancerous groups due to the picrocrocin treatment in comparison with control and cancerous groups, without treatment. In addition, tumor size decreased in cancerous group with picrocrocin treatment. In conclusion, our results indicated the beneficial effect of picrocrocin in breast cancer treatment. Keywords: Saffron, Breast cancer, Picrocrocin, Enkephalin degrading enzyme
doi:10.1016/j.clinbiochem.2011.08.784
Poster – [A-10-1096-1] Effect of glycerol, as a chemical chaperone, on hemoglobin glycation by glucose or fructose Asghar Farajzadea, S. Zahra Bathaieb a Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat ModaresIran University, Tehran, Iran b Iran, Tehran, Tarbiat Modares University, Faculty of Medical Sciences, Department of Clinical Biochemistry.P.O.BOX: 14114–111 E-mail addresses: [email protected] (A. Farajzade), [email protected] (S.Z. Bathaie) Introduction: None enzymatic reactions between reducing sugars and proteins produce some complexes, which are named as advanced glycation end products (AGEs). They can play an important role in diabetic complications, cancer, atherosclerosis, aging, prion and Alzheimer diseases, etc. Protocols that result in the decrease of AGEs can help people faced with these diseases. There are some methods to decrease AGEs like controlling carbohydrate`s concentration in the body or using some drugs such as metformin, aspirin and so on. Here, we used chemical chaperones for this purpose. Chemical chaperons are molecules with low molecular weight which are known for the body and deal with proteins to stabilize them against thermal or chemical denaturation. Methods: Glycerol, as a polyol, is selected to prevent hemoglobin glycation by glucose or fructose. Hemoglobin solution, pH 7.4, is incubated separately with glucose or fructose in the presence or absence of glycerol at 37 °C in a shaker incubator. After completion of the reaction, all samples are investigated by different techniques. Results: Incubated Hb with both glucose/fructose and glycerol shows less fluorescence emission in comparison with solution containing glycerol. In addition, there are some changes in the electrophretic
mobility and secondary structure content of hemoglobin in comparison with Hb+ glucose/fructose in the presence or absence of glycerol. Conclusion: AGE formation causes an increase in the fluorescence emission, a decrease in the alpha-helix content and an increase in the electrophoretic mobility of Hb. However, glycerol has preventing effect on these changes and stabilizes hemoglobin against reducing sugars. Keywords: Glycerol, Chemical chaperon, Hemoglobin, AGE, Glycation doi:10.1016/j.clinbiochem.2011.08.785
Poster — [A-10-1101-1] Effects of diabetes on myocardial capillary density and serum biomarkers of angiogenesis in male rats Khazaei Majid, Fallahzadeh Alireza Department of Physiology, Isfahan University of Medical Sciences, Hezar Jarib Ave, Isfahan, Iran E-mail addresses: [email protected] (K. Majid), [email protected] (F. Alireza) Introduction: Cardiovascular disease is one of the main causes of mortality and morbidity in diabetic subjects. This study evaluates the effect of diabetes on myocardial capillary density and some serum angiogenic factors including nitric oxide, vascular endothelial growth factor and its soluble receptors. Methods: Twelve male rats were divided into two groups; control and diabetic (n = 6 each). Diabetes was induced by single dose of streptozotocin (50 mg/kg, intraperitoneal). After 21 days, capillary density in myocardial tissue was evaluated by immunohistochemical staining and reported as capillaries per 2 mm. Blood samples were collected before and after experiment. Results: In diabetic group, serum nitric oxide and soluble vascular endothelial growth factor receptor-2 concentrations were lower and soluble vascular endothelial growth factor receptor-1 was significantly higher than the control group. There was no significant change in serum vascular endothelial growth factor concentration between diabetic and control groups, however, vascular endothelial growth factor/vascular endothelial growth factor receptor-1 ratio was significantly decreased in diabetic animals. Myocardial capillary density in diabetic group was lower than the control group (1549 ± 161 vs. 2156 ± 202/2 mm, respectively). Conclusion: Reduced serum nitric oxide and vascular endothelial growth factor receptor-2 levels, increased serum vascular endothelial growth factor receptor-1 and lower vascular endothelial growth factor/ vascular endothelial growth factor receptor-1 ratio might be responsible for decreased myocardial capillary density in diabetic rats. Keywords: Diabetes, Capillary density, Vascular endothelial growth factor, Nitric oxide, Myocardium doi:10.1016/j.clinbiochem.2011.08.929
Poster — [A-10-1107-1] Study of the effect of glucose on spectroscopy features of doxorubicin anticancer drug Abri Mehraban Fatemeh, Zargar Seyed Jalal, Habibi Rezayee Mehran University of Tehran, Islamic Republic of Iran E-mail addresses: [email protected] (A.M. Fatemeh), [email protected] (Z.S. Jalal), [email protected] (H.R. Mehran) Introduction: Doxorubicin is an anthracycline antibiotic being used in chemotherapy of a wide range of different cancers specifically solid
Abstracts
tumors and leukemia. Glucose is the most common hexose carbohydrate which has specific metabolically roles in biological systems. Method: In this study the interaction of doxorubicin with different glucose concentrations (100, 300 and 500 mg/dl) at room temperature was investigated for 2 h, using ultraviolet spectroscopy technique (200– 600 nm). Results: The results showed that glucose increases the absorbance of doxorubicin at the wavelength of 480 nm and induces hyperchromicity in the absorption spectrum of this drug. Conclusion: This study demonstrated that glucose affects doxorubicin and changes its absorbance at 480 nm, and UV spectra. Since the absorbance at 480 nm corresponds to tetracycline ring of doxorubicin, the results suggested that glucose probably interacts with the tetracycline group of drug molecule.
S179
Keywords: Simvastatin therapy, Prooxidant–antioxidant Balance, Heat shock protein 60, 65, and 70 doi:10.1016/j.clinbiochem.2011.08.931
Poster — [A-10-1141-1] The effect of different concentrations of morphine and extracellular Ca2+ on cell death in PC12 cell line Arezou Rabziaa, Mehri Azadbakhta, Ali Bidmeshki Poura, Hasan Akramia a Department of Biology, Razi University, Kermanshah, Iran E-mail addresses: [email protected] (A. Rabzia), [email protected] (M. Azadbakht)
Keywords: Doxorubicin, Glucose, UV spectroscopy doi:10.1016/j.clinbiochem.2011.08.930
Poster — [A-10-1123-1] Simvastatin therapy reduces prooxidant–antioxidant balance and heat shock protein 60, 65, and 70 antibody titers: Results of a placebo-controlled cross-over trial Barzegar Amini Maral, Mouhebati Mohsen, Ghayour-Mobarhan Majid, Azarpazhooh Mohammad-Reza, Sahebkar Amir-Hosein, Hassanzade-Daloee Mahdi, Momenzade Akram, Tavallaei Shima, Rahsepar Amir-Ali, Khojasteh-Taheri Roshanak Mashhad University of Medical Siences, Mashhad, Iran E-mail addresses: [email protected] (B.A. Maral), [email protected] (M. Mohsen), [email protected] (G.-M. Majid), [email protected] (A. Mohammad-Reza), [email protected] (S. Amir-Hosein), [email protected] (H.-D. Mahdi), [email protected] (M. Akram), [email protected] (T. Shima), [email protected] (R. Amir-Ali), [email protected] (K.-T. Roshanak) Introduction: Oxidative stress and heat shock proteins (HSPs) are thought to play an important role in atherogenesis. The statin group of cholesterol-lowering drugs has been shown to reduce cardiovascular events and possess antioxidant properties and immunomodulatory effects. This study was carried out to evaluate the effects of statin therapy on serum levels of antibodies HSPs and prooxidant– antioxidant balance (PAB), as novel coronary risk factors in dyslipidemic patients. Method: The PAB assay can measure the prooxidant burden and the antioxidant capacity simultaneously in one assay, thereby giving a redox index. We treated 102 dyslipidemic individuals with simvastatin, or a placebo in a double-blind, cross-over, placebo-controlled trial. PAB values and Anti-HSP60, 65, 70, and hs-CRP levels were measured before and after each treatment period. Seventy-seven subjects completed the study. Result: We found that statin therapy was associated with a significant reduction in PAB values (P < 0.001) and serum anti-HSP60, 65, and 70 titers in the dyslipidemic patients (10%, 14%, and 15% decrease, respectively) (p < 0.001). There have been previous reports of reductions in serum CRP with statin treatment, and although median CRP levels were 9% lower on simvastatin treatment, this did not achieve statistical significance. Conclusions: These findings suggest that simvastatin inhibits autoimmune responses and oxidative stress that may contribute to the development of cardiovascular disease.
Introduction: Morphine as a mu opioid drug can induce different effects. Ca2+ as one of the key regulators of cell survival, in response to a wide range of conditions, is capable of inducing cell death. Here, we investigated the effect of different concentrations of morphine together with different concentrations of extracellular Ca2+ on cell death in PC12 cell line. Materials and methods: PC12 cells were cultured in RPMI 1640 culture medium containing 10% fetal bovine serum and different concentrations of morphine (10 E-6 and 10 E-4 M) together with different concentrations of extracellular Ca2+ (0.7, 0.6, 0.5, 0.4, 0.3, 0.2 and 0.00 mM) for 6, 12 and 24 h. PC12 cells that were cultured without morphine were considered as control. Cell viability was assessed by neutral red staining. We used Hoechst/PI nuclear staining and fluorescence microscopy for detection of percentage of apoptotic and necrotic cells. Results: Results indicated that cell viability of PC12 cells in the presence of concentration of 10 E-4 M of morphine together with 0.7 mM of extracellular Ca2+ decreased compared to control (P < 0.05). Hoechst/PI nuclear staining demonstrated that 10 E-4 M of morphine together with 0.7 mM of extracellular Ca2+ were toxic and increased the percentage of apoptotic and necrotic cells. Conclusion: It is showed that high-dose of morphine decreasesd cell viability and enhanced apoptosis and necrosis in PC12 cells, that was dependent on the presence of extracellular Ca2+. Keywords: Morphine, Ca2+, Cell death, Apoptosis, PC12 cell
doi:10.1016/j.clinbiochem.2011.08.932
Poster — [A-10-1147-1] Evaluation of anticoagulant properties of Artemisia dracunculus, Punica granatum and Berberis vulgaris in rat Hosseinzadeh Elyas, Razieh University of Tehran, I.B.B, Laboratory of Genetic Engineering, Islamic Republic of Iran E-mail addresses: [email protected] (H. Elyas), [email protected] (Razieh) Introduction: One of the most important pathways leading to thrombosis is platelet adhesion to extracellular matrix proteins that in turn causes platelet plug formation and eventually formation of blood clot (thrombus) inside a blood vessel that leads to cardiovascular desease. Punica granatum is used as blood dilutor and antidiabetic effect, and Artemisia dracunculus and Berberis vulgaris are used as an anticoaguant and antihyperlipidaemic agents in Iranian folk medicine. In that line, we investigated the inhibitory effects of crude extracts of these plants on the adhesion of the activated platelets to fibrinogen coated plates.