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An animal model of postoperative ileus
202
Abstracts
Cardiotoxicity is one of the major causes of failed drug development and withdrawals. Over-expressed hERG and other cardiac ion channels screening assays are the major in vitro assays to predict cardiotoxicity. However, the ability of ion channel screening assays to accurately predict clinical cardiotoxic outcomes is less than satisfactory. Three technologies were used in this study to develop comprehensive and predictive cardiotoxicity assays using human stem cell derived cardiomyocytes (hESC-CM): multi-electrode array (MEA), conventional patch clamp and high content analysis (HCA). The MEA measuring cardiac field potential parameters detects the compounds that disrupt electrical signal conduction. The conventional patch clamp measuring action potential parameters confirms MEA results. The HCA measuring cell health parameters detects the compounds that cause structural damage, which may not be identified by electrophysiological methods. The MEA assay showed significant increase of field potential duration by E-4031 and Sotalol; and significant decrease in field potential duration by Nifedipine and Isoproterenol. The conventional patch clamp assay showed significant increase in action potential duration at 60% and 90% repolarization for Sotalol and Quinidine, but significant decrease for Nifedipine. HCA assays measure cell count, calcium mobilization, mitochondrial membrane potential (MMP), membrane permeability, B-type natriuretic peptide, alpha-actinin disruption and troponin I integrity. Twelve drugs were incubated for 24 and 72 h. IC50 values demonstrated high concordance with published values. More adverse effects were observed at 72 h. MMP was the most sensitive and specific indicator of structural cardiotoxicity. This study demonstrates using hESC-CM by MEA and HCA can facilitate cardiotoxicity identification. doi:10.1016/j.vascn.2015.08.146
0149 Renal safety pharmacology: Characterization of a non-rodent model with water loading using a positive control agent Samir Abtouta, Alexis Ascaha, Leanne Bassetta,b, Mytlene Pouliota, Simon Authiera,b a
CiToxLAB, Laval, Quebec, Canada Université de Montréal, St-Hyacinthe, Canada
b
Introduction: Renal safety pharmacology is commonly performed in rodents but limited data is available for non-rodents. A canine model of renal safety pharmacology with water loading was characterized. Methods: Male beagle dogs (n = 6) received furosemide (0.3, 3 and 10 mg/kg) or control in a crossover design with a 7 day washout. Water loading (30 ml/kg) and urinary bladder emptying were completed prior to each treatment. Each treatment session was divided into baseline (0–6 h and 6–24 h) and post-dose collections (0–2 h, 2–4 h, 4–6 h and 6–24 h). Biochemistry was evaluated at 6 h and 24 h post-dose. Urine volume (ml/kg), specific gravity, plasma and urine biochemical parameters, water consumption and food consumption were assessed. Glomerular filtration rate, water clearance and excretion fraction were calculated to evaluate renal function. Results: Water loading increased urine output and glomerular filtration rate (0–6 h). Furosemide (0.5, 3 and 10 mg/kg) induced a dose dependent increase in urine output at 0–6 h. The 24 h urine volume was moderately increased (+60.7%) with furosemide at 10 mg/kg compared to control. From 0–6 h, furosemide increased free water clearance, urinary excretion of sodium, calcium and chloride, decreased urine osmolarity and specific gravity. Water consumption increased and was proportional to urine output while food consumption decreased slightly at high dose (10 mg/kg) only.
Conclusion: Renal safety pharmacology in a canine model with water loading enabled identification of pharmacodynamic effects following administration of a diuretic. The canine model represents a valid alternative when this species is justified. doi:10.1016/j.vascn.2015.08.147
0150 Evaluation of role of atorvastatin in cisplatin induced nephrotoxicity in Wistar rats Sravan Kumar Boorla Vaagdevi College of Pharmacy, Warangal, Andhra Pradesh, India Objectives: This study was carried out to investigate the ameliorative effect of atorvastatin on cisplatin-induced nephrotoxicity in Wistar rats. Methods: Twenty-four (24) male rats were used for the experiment. Group I was administered per oral daily with phosphate buffered saline (PBS) for 10 days. Group II was administered single dose of cisplatin 7.5 mg/kg (ip) on day five, group III was administered daily per oral dose of atorvastatin 3 mg/kg for 10 days and day five received a single dose of cisplatin 7.5 mg/kg (ip). Group IV was administered daily per oral dose of atorvastatin 10 mg/kg for 10 days and on day five received a single dose of cisplatin 7.5 mg/kg (ip). Following the last treatment, blood samples were collected by cardiac puncture for analysis of serum creatinine, blood urea nitrogen and total protein. The antioxidant parameters catalase, GSH and TBARS were measured in kidneys. Results: Groups treated with cisplatin only showed a significant reduction in creatinine, urea and total protein relative to PBS-treated group. Conclusion: The observed decreases in creatinine, blood urea and total protein were ameliorated by administration of atorvastatin which may be due to its antioxidant properties. doi:10.1016/j.vascn.2015.08.148
0151 An animal model of postoperative ileus Sonia Goineau, Phillipe Guillaume, Marion Hunault, Sylvie Bézivin, Guillaume Froget Porsolt SAS, Le Genest St Isle, France Postoperative ileus (POI) is a transient impairment of gastrointestinal functions developing as a consequence of abdominal surgery. POI was induced in the mouse by gentle manipulation of the small intestine. Approximately 24 h after intestinal manipulation (IM) or sham procedure, the effects of mosapride (a gastroprokinetic agent), or vehicle, on intestinal transit were investigated by evaluating the distribution along the gastrointestinal tract of phenol red. Mosapride was administered 1 h before IM, 6 h after IM and then, 1 h before test meal at 20 mg/kg each time. In sham animals, 78% of the phenol red test meal was transported down the intestine up to the distal end of colon. In IM animals, the distribution of phenol red was evident towards the more proximal segments of the gastrointestinal tract, a finding which is consistent with the presence of POI. Intestinal transit was reduced in IM animals as compared to sham controls (geometric center: 2.34 ± 0.07 versus 5.22 ± 0.40 in sham,
Abstracts
p b 0.001). No significant difference was observed in the gastric emptying. However, fecal output was decreased in IM animals (99.0 ± 11.3 mg versus 158.9 ± 11.5 mg of stools in sham, p b 0.01). Mosapride accelerated intestinal transit (geometric center: 3.50 ± 0.34 versus 2.34 ± 0.07, p b 0.01) and gastric emptying (85.3 ± 5.6% versus 68.6 ± 3.3%, p b 0.05) as compared to IM control group. Mosapride also tended to show a slight increase in fecal
203
output. In conclusion, mosapride attenuated IM-induced intestinal motility dysfunction by increasing gastric emptying and gastrointestinal transit, validating the use of this model for POI.
doi:10.1016/j.vascn.2015.08.149
Abstracts
Cardiotoxicity is one of the major causes of failed drug development and withdrawals. Over-expressed hERG and other cardiac ion channels screening assays are the major in vitro assays to predict cardiotoxicity. However, the ability of ion channel screening assays to accurately predict clinical cardiotoxic outcomes is less than satisfactory. Three technologies were used in this study to develop comprehensive and predictive cardiotoxicity assays using human stem cell derived cardiomyocytes (hESC-CM): multi-electrode array (MEA), conventional patch clamp and high content analysis (HCA). The MEA measuring cardiac field potential parameters detects the compounds that disrupt electrical signal conduction. The conventional patch clamp measuring action potential parameters confirms MEA results. The HCA measuring cell health parameters detects the compounds that cause structural damage, which may not be identified by electrophysiological methods. The MEA assay showed significant increase of field potential duration by E-4031 and Sotalol; and significant decrease in field potential duration by Nifedipine and Isoproterenol. The conventional patch clamp assay showed significant increase in action potential duration at 60% and 90% repolarization for Sotalol and Quinidine, but significant decrease for Nifedipine. HCA assays measure cell count, calcium mobilization, mitochondrial membrane potential (MMP), membrane permeability, B-type natriuretic peptide, alpha-actinin disruption and troponin I integrity. Twelve drugs were incubated for 24 and 72 h. IC50 values demonstrated high concordance with published values. More adverse effects were observed at 72 h. MMP was the most sensitive and specific indicator of structural cardiotoxicity. This study demonstrates using hESC-CM by MEA and HCA can facilitate cardiotoxicity identification. doi:10.1016/j.vascn.2015.08.146
0149 Renal safety pharmacology: Characterization of a non-rodent model with water loading using a positive control agent Samir Abtouta, Alexis Ascaha, Leanne Bassetta,b, Mytlene Pouliota, Simon Authiera,b a
CiToxLAB, Laval, Quebec, Canada Université de Montréal, St-Hyacinthe, Canada
b
Introduction: Renal safety pharmacology is commonly performed in rodents but limited data is available for non-rodents. A canine model of renal safety pharmacology with water loading was characterized. Methods: Male beagle dogs (n = 6) received furosemide (0.3, 3 and 10 mg/kg) or control in a crossover design with a 7 day washout. Water loading (30 ml/kg) and urinary bladder emptying were completed prior to each treatment. Each treatment session was divided into baseline (0–6 h and 6–24 h) and post-dose collections (0–2 h, 2–4 h, 4–6 h and 6–24 h). Biochemistry was evaluated at 6 h and 24 h post-dose. Urine volume (ml/kg), specific gravity, plasma and urine biochemical parameters, water consumption and food consumption were assessed. Glomerular filtration rate, water clearance and excretion fraction were calculated to evaluate renal function. Results: Water loading increased urine output and glomerular filtration rate (0–6 h). Furosemide (0.5, 3 and 10 mg/kg) induced a dose dependent increase in urine output at 0–6 h. The 24 h urine volume was moderately increased (+60.7%) with furosemide at 10 mg/kg compared to control. From 0–6 h, furosemide increased free water clearance, urinary excretion of sodium, calcium and chloride, decreased urine osmolarity and specific gravity. Water consumption increased and was proportional to urine output while food consumption decreased slightly at high dose (10 mg/kg) only.
Conclusion: Renal safety pharmacology in a canine model with water loading enabled identification of pharmacodynamic effects following administration of a diuretic. The canine model represents a valid alternative when this species is justified. doi:10.1016/j.vascn.2015.08.147
0150 Evaluation of role of atorvastatin in cisplatin induced nephrotoxicity in Wistar rats Sravan Kumar Boorla Vaagdevi College of Pharmacy, Warangal, Andhra Pradesh, India Objectives: This study was carried out to investigate the ameliorative effect of atorvastatin on cisplatin-induced nephrotoxicity in Wistar rats. Methods: Twenty-four (24) male rats were used for the experiment. Group I was administered per oral daily with phosphate buffered saline (PBS) for 10 days. Group II was administered single dose of cisplatin 7.5 mg/kg (ip) on day five, group III was administered daily per oral dose of atorvastatin 3 mg/kg for 10 days and day five received a single dose of cisplatin 7.5 mg/kg (ip). Group IV was administered daily per oral dose of atorvastatin 10 mg/kg for 10 days and on day five received a single dose of cisplatin 7.5 mg/kg (ip). Following the last treatment, blood samples were collected by cardiac puncture for analysis of serum creatinine, blood urea nitrogen and total protein. The antioxidant parameters catalase, GSH and TBARS were measured in kidneys. Results: Groups treated with cisplatin only showed a significant reduction in creatinine, urea and total protein relative to PBS-treated group. Conclusion: The observed decreases in creatinine, blood urea and total protein were ameliorated by administration of atorvastatin which may be due to its antioxidant properties. doi:10.1016/j.vascn.2015.08.148
0151 An animal model of postoperative ileus Sonia Goineau, Phillipe Guillaume, Marion Hunault, Sylvie Bézivin, Guillaume Froget Porsolt SAS, Le Genest St Isle, France Postoperative ileus (POI) is a transient impairment of gastrointestinal functions developing as a consequence of abdominal surgery. POI was induced in the mouse by gentle manipulation of the small intestine. Approximately 24 h after intestinal manipulation (IM) or sham procedure, the effects of mosapride (a gastroprokinetic agent), or vehicle, on intestinal transit were investigated by evaluating the distribution along the gastrointestinal tract of phenol red. Mosapride was administered 1 h before IM, 6 h after IM and then, 1 h before test meal at 20 mg/kg each time. In sham animals, 78% of the phenol red test meal was transported down the intestine up to the distal end of colon. In IM animals, the distribution of phenol red was evident towards the more proximal segments of the gastrointestinal tract, a finding which is consistent with the presence of POI. Intestinal transit was reduced in IM animals as compared to sham controls (geometric center: 2.34 ± 0.07 versus 5.22 ± 0.40 in sham,
Abstracts
p b 0.001). No significant difference was observed in the gastric emptying. However, fecal output was decreased in IM animals (99.0 ± 11.3 mg versus 158.9 ± 11.5 mg of stools in sham, p b 0.01). Mosapride accelerated intestinal transit (geometric center: 3.50 ± 0.34 versus 2.34 ± 0.07, p b 0.01) and gastric emptying (85.3 ± 5.6% versus 68.6 ± 3.3%, p b 0.05) as compared to IM control group. Mosapride also tended to show a slight increase in fecal
203
output. In conclusion, mosapride attenuated IM-induced intestinal motility dysfunction by increasing gastric emptying and gastrointestinal transit, validating the use of this model for POI.
doi:10.1016/j.vascn.2015.08.149