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Inhibition of collagen- and ADP-induced platelet aggregation by natural products
HAVE FLAVONOIDS
EFFECT ON THE AFFINITk
ON NATIVE LDL TO ITS RECEPTOR? Seddigheh. kary? Gholam.iui. Naderi, Mohsen Am. SarraEadegaR Roya. Klishadi, Mohammad.Reza. Safari
Nizal.
Isfian Cardiovascular Research Confer, Isfahan, Iran. OBJECTIVES: Flavonoids are phenolic compounds widely distributed in plants and the antioxidative effect of these compounds has been of mterest for a considerable time. We studied the effects of smne flavonoids including Apigenin, Geniti and Pelargomdin chloride on the atTmity of native LDL to its receptor. METHODS: First, LDL was isolated from normal serum using density ultracentiitigation and, then was labeled with Flourescin iso thiocyanate (FITC) solution (in bicarbonate buffer with uH=8.5) at 4’C for 24hr. The LDL receptor was purified of bovine adrenal t&e. These flavonoids dissolved in DMSO (Dimethyl sulfoxide). After 20min mcubatmn of native LDL with various concentrations of these flavonoids, the LDL receptor and labeled-LDL were incubated with them for 30min at 37’C. so that, final concentration of flavonoids for each sample were 50, 100 and 200wM
The medium was centrifuged (4OOOg for 20min) and
supematant was isolated and then intensity of the fluorescence was measured wth an excitation at 495nm and an ermssion at 5 15nm. RESULTS: Results shown that these flavonoids were increased the aiTmity of LDL to its receptor. The effects of these flavonoids is respectively Genistin> PelargonidinChloride> Apigenin. Conclusions: These results demonstrated that flavonoids probably can prevent the destructive effects of LDL. T: These flavonoids significantly inhibit in vitro LDL oxidation in the order of Biocbanii A>Apigenin>Genistin>Morin COCULUSION: This study show that flavonoids prevent in vitro LDL oxidation and probably would be unportant to prevent atherosclerosis.
NICOTINE AND COTININE ENHANCED THE HEMOGLOBIN GLY COSYLATION A: POSSIBLE DELETERIOUS MECHANISM OF SMOKING Seddtgheh. Asgaty, Gholam Ah. Naderi, Nizal. Sarrafzadegan, Maryam Boshtam, Motteza Rafei, Maryam zarei, Soma. Zxfeshany, Isfahan Cardiovascular Research Center, Isfahan, Iran Objective: The non enzymatic glycosylation of proteins in vlvo may lead to various complications that it is important especially in generatmg a series of pathogenesis in diabetic patients. Free ammo group m protein react with acyclic form of glucose to a destructive compound without its physiological effects and even wth harmful actions. This study IS mmed to find the rtrects of both tobacco mcotme and Its metabohte cotmine on hemoglobm glycosylation m vitro. METHOD: At first the best concentration and the best time to incubate glucose with hemoglobin was investigated Then the glycosylation degree of hemoglobin in the prescence and absence of nicotine and cotmine was measured by means of spectrophotometric method Different con&rations of mcotme and cotinine were used. RESULT: Nicotine at final concentration of 12mg/ml and 25mg/ml increases the rate of glycosylatlon 30.7% , and 47 9% respectwely and cotimne by O.O13mg/ml, 0 Zmg/ml and 0 3 mg/ml concentrations increases the glycosylation degree 8 l%, 10.6% and 12.3% respectively CONCLUSION: Therefore promolmg mechanisms of hemoglobin glycosylation 1s one of the alternatives for increasmg risk of various diseases in diabetx and healthy people who smoke.
A6
INHIBITION OF COLLAGENAND ADP-INDUCED PLATELET AGGREGATION BY NATURAL PRODUCTS Seddigheh Asgaty, Ghohn-Ali Naderi, Nizal Sarrafiadgau, Mojgan Gharipour, lsfahan Cardiovascular Research Center, lsfahan, Iran PURPOSE: Thrombosis plays an especially important role in the causes of cardiac diseases. The antiplatelet effects of some Iranian herbel medicine (Ribes orientale, cornus mas, Fumaria o!Xcinale) that used traditionally as anti-coagulant drug studied on the aggregation of human platelet-rich plasma induced by collagen and ADP. METHOD: Polyphenolic extract of each plant was prepared and dissolved in dimethyl sulfoxide (DMSO). Platelet-numbers were counted by Coulter Counter. The platelet were suspended in Tyxde’s solution. Aggregation was measured by a turbidimetric method using Lumiaggregometer. The final concentration of each plant extract was Sp/ml. Aspirin was used as a positive control. RESULT: The result show that Ribes orientale, cornus mas, Fumwia officinale inhibited ADP-induced platelet aggregation resepectively 70%, 60% and 45%. These extract also inhibited collagen-induced platelet aggregation respectively 55% , 60% , 53%. Also, Aspirin completely inhibited the aggregation mduced by collagen or ADP. CONCLUSIONS: This study confirms that the three plants used in traditional medication in Jran that used as anticoagulants. Also, by carrying out vast studies, these plants with their extracted materials can be used in the treatment or prevention of cardiovascular disease.
TRANSCRIPTIONAL RESPONSES TO ISCHEMIA IN AlAR TRANSGENIC AND WILD-TYPE HEARTS Kevin Ashton, Klrsty Holmgren, Sean Grimmond, Paul Matherne & John Headrlck. Heart Foundation Research Centre, Griffith University-Gold Coast, QLD, Australia. K.Ashton@mai/box.gu.edu.au It is well established that adenosine receptor (AR) activation reduces myocardial infarction and post-ischemic stunning. Our previous studies using an AlAR transgenic overexpression model demonstrate increased ischemic tolerance compared with wild-type hearts. At present very little is known about the underlying mechanisms of this cardioprotection, or regarding molecular mechanisms of ischemic injury. We have examined the transcriptional consequences of ischemia using a cDNA microarray approach. Wild-type control and AlAR transgenic murine hearts were investigated under normoxic and postischemic conditions. Briefly, isolated Langendorff perfused hearts were subjected to 30 minutes ischemia, 60 minutes reperfusion, total RNA extracted and pooled (n=6/group) for each group. Fluorescently labelled RNA was then cohybridised with a wild-type normoxic control to custom cDNA microarrays. We have identified differentially regulated transcripts in response to ischemia-reperfusion, particularly for metabolic enzymes, cytoskeletal/structural proteins and other novel transcripts. The pattern of expression is substantially modified by AlAR overexpression, which also markedly enhanced ischemic tolerance. Cardioprotection with AlAR overexpression may involve reversal of down-regulation of metabolic enzymes and cytoskeletallstructural proteins. Our findings are presented and discussed in light of known genetic and biological consequences of ischemic injury.
EFFECT ON THE AFFINITk
ON NATIVE LDL TO ITS RECEPTOR? Seddigheh. kary? Gholam.iui. Naderi, Mohsen Am. SarraEadegaR Roya. Klishadi, Mohammad.Reza. Safari
Nizal.
Isfian Cardiovascular Research Confer, Isfahan, Iran. OBJECTIVES: Flavonoids are phenolic compounds widely distributed in plants and the antioxidative effect of these compounds has been of mterest for a considerable time. We studied the effects of smne flavonoids including Apigenin, Geniti and Pelargomdin chloride on the atTmity of native LDL to its receptor. METHODS: First, LDL was isolated from normal serum using density ultracentiitigation and, then was labeled with Flourescin iso thiocyanate (FITC) solution (in bicarbonate buffer with uH=8.5) at 4’C for 24hr. The LDL receptor was purified of bovine adrenal t&e. These flavonoids dissolved in DMSO (Dimethyl sulfoxide). After 20min mcubatmn of native LDL with various concentrations of these flavonoids, the LDL receptor and labeled-LDL were incubated with them for 30min at 37’C. so that, final concentration of flavonoids for each sample were 50, 100 and 200wM
The medium was centrifuged (4OOOg for 20min) and
supematant was isolated and then intensity of the fluorescence was measured wth an excitation at 495nm and an ermssion at 5 15nm. RESULTS: Results shown that these flavonoids were increased the aiTmity of LDL to its receptor. The effects of these flavonoids is respectively Genistin> PelargonidinChloride> Apigenin. Conclusions: These results demonstrated that flavonoids probably can prevent the destructive effects of LDL. T: These flavonoids significantly inhibit in vitro LDL oxidation in the order of Biocbanii A>Apigenin>Genistin>Morin COCULUSION: This study show that flavonoids prevent in vitro LDL oxidation and probably would be unportant to prevent atherosclerosis.
NICOTINE AND COTININE ENHANCED THE HEMOGLOBIN GLY COSYLATION A: POSSIBLE DELETERIOUS MECHANISM OF SMOKING Seddtgheh. Asgaty, Gholam Ah. Naderi, Nizal. Sarrafzadegan, Maryam Boshtam, Motteza Rafei, Maryam zarei, Soma. Zxfeshany, Isfahan Cardiovascular Research Center, Isfahan, Iran Objective: The non enzymatic glycosylation of proteins in vlvo may lead to various complications that it is important especially in generatmg a series of pathogenesis in diabetic patients. Free ammo group m protein react with acyclic form of glucose to a destructive compound without its physiological effects and even wth harmful actions. This study IS mmed to find the rtrects of both tobacco mcotme and Its metabohte cotmine on hemoglobm glycosylation m vitro. METHOD: At first the best concentration and the best time to incubate glucose with hemoglobin was investigated Then the glycosylation degree of hemoglobin in the prescence and absence of nicotine and cotmine was measured by means of spectrophotometric method Different con&rations of mcotme and cotinine were used. RESULT: Nicotine at final concentration of 12mg/ml and 25mg/ml increases the rate of glycosylatlon 30.7% , and 47 9% respectwely and cotimne by O.O13mg/ml, 0 Zmg/ml and 0 3 mg/ml concentrations increases the glycosylation degree 8 l%, 10.6% and 12.3% respectively CONCLUSION: Therefore promolmg mechanisms of hemoglobin glycosylation 1s one of the alternatives for increasmg risk of various diseases in diabetx and healthy people who smoke.
A6
INHIBITION OF COLLAGENAND ADP-INDUCED PLATELET AGGREGATION BY NATURAL PRODUCTS Seddigheh Asgaty, Ghohn-Ali Naderi, Nizal Sarrafiadgau, Mojgan Gharipour, lsfahan Cardiovascular Research Center, lsfahan, Iran PURPOSE: Thrombosis plays an especially important role in the causes of cardiac diseases. The antiplatelet effects of some Iranian herbel medicine (Ribes orientale, cornus mas, Fumaria o!Xcinale) that used traditionally as anti-coagulant drug studied on the aggregation of human platelet-rich plasma induced by collagen and ADP. METHOD: Polyphenolic extract of each plant was prepared and dissolved in dimethyl sulfoxide (DMSO). Platelet-numbers were counted by Coulter Counter. The platelet were suspended in Tyxde’s solution. Aggregation was measured by a turbidimetric method using Lumiaggregometer. The final concentration of each plant extract was Sp/ml. Aspirin was used as a positive control. RESULT: The result show that Ribes orientale, cornus mas, Fumwia officinale inhibited ADP-induced platelet aggregation resepectively 70%, 60% and 45%. These extract also inhibited collagen-induced platelet aggregation respectively 55% , 60% , 53%. Also, Aspirin completely inhibited the aggregation mduced by collagen or ADP. CONCLUSIONS: This study confirms that the three plants used in traditional medication in Jran that used as anticoagulants. Also, by carrying out vast studies, these plants with their extracted materials can be used in the treatment or prevention of cardiovascular disease.
TRANSCRIPTIONAL RESPONSES TO ISCHEMIA IN AlAR TRANSGENIC AND WILD-TYPE HEARTS Kevin Ashton, Klrsty Holmgren, Sean Grimmond, Paul Matherne & John Headrlck. Heart Foundation Research Centre, Griffith University-Gold Coast, QLD, Australia. K.Ashton@mai/box.gu.edu.au It is well established that adenosine receptor (AR) activation reduces myocardial infarction and post-ischemic stunning. Our previous studies using an AlAR transgenic overexpression model demonstrate increased ischemic tolerance compared with wild-type hearts. At present very little is known about the underlying mechanisms of this cardioprotection, or regarding molecular mechanisms of ischemic injury. We have examined the transcriptional consequences of ischemia using a cDNA microarray approach. Wild-type control and AlAR transgenic murine hearts were investigated under normoxic and postischemic conditions. Briefly, isolated Langendorff perfused hearts were subjected to 30 minutes ischemia, 60 minutes reperfusion, total RNA extracted and pooled (n=6/group) for each group. Fluorescently labelled RNA was then cohybridised with a wild-type normoxic control to custom cDNA microarrays. We have identified differentially regulated transcripts in response to ischemia-reperfusion, particularly for metabolic enzymes, cytoskeletal/structural proteins and other novel transcripts. The pattern of expression is substantially modified by AlAR overexpression, which also markedly enhanced ischemic tolerance. Cardioprotection with AlAR overexpression may involve reversal of down-regulation of metabolic enzymes and cytoskeletallstructural proteins. Our findings are presented and discussed in light of known genetic and biological consequences of ischemic injury.