- Email: [email protected]
Marmoset monkeys as preclinical models for respiratory diseases
S204
Abstracts / Toxicology Letters 196S (2010) S37–S351
P205-037 Marmoset monkeys as preclinical models for respiratory diseases S. Seehase 1 , H.D. Lauenstein 1 , S. Switalla 1 , F. Prenzler 1 , F.J. Kaup 2 , E. Fuchs 2 , N. Krug 1 , C. Schlumbohm 2 , K. Sewald 1 , A. Braun 1 1
Fraunhofer Institute for Toxicology and Experimental Medicine, Germany, 2 German Primate Center, Germany Therapeutic efficacy studies of biopharmaceuticals for respiratory diseases are often limited in laboratory animals due to high interspecies differences. In order to adapt preclinical models closer to the human situation we are currently establishing lipopolysaccharide (LPS)-induced inflammation models in a non-human primate, the marmoset monkey (Callithrix jacchus). Whole blood (WB) samples were incubated with LPS in vitro to determine the individual immunological potential of the animals. Precision cut lung slices (PCLS) were used to investigate the immunological potential in the target organ lung. Dexamethasone served as treatment control in both, PCLS and WB-assay. In an in vivo approach marmosets were intrabronchially exposed to LPS. Before LPS instillation a unilateral bronchoalveolar lavage (BAL) was performed in anaesthetized animals under bronchoscopical supervision to determine BAL cell counts and TNF-alpha release at baseline levels. LPS was instilled in anesthetized animals intratracheally into the left lung via microsprayer. Thereafter a second BAL was performed 18 h later. Cell counts and TNF-alpha concentration were determined in BAL fluid. To monitor systemic effects, haematology, body weight and TNF-alpha concentration in serum were analyzed before and after LPS provocation. Both, WB-assay and PCLS showed a dose-dependent TNF-alpha response to LPS. Significant correlations were found between PCLS and WB-assay (r2 = 0.9). The same was true for human and marmoset PCLS (r2 = 0.9). Dexamathasone treatment reduced TNFalpha release significantly (WBA to 59%, PCLS to 77%). In the in vivo approach LPS instillation caused a moderate TNF-alpha increase (<5 pg/ml vs. 28 pg/ml) and a strong neutrophilic influx in BAL fluid (<1% vs. 79% after LPS), approved by histology and BAL cell counts. No signs of systemic inflammation were detectable. In conclusion, marmoset monkeys represent a suitable model to study inflammatory effects in the lung and their therapeutical intervention in vitro, ex vivo, and in vivo, close to the human situation. doi:10.1016/j.toxlet.2010.03.688
P206 Target Organ Toxicity. Hepatotoxicity P206-001 Beneficial effect of combined administration of some naturally occurring antioxidants and thiol chelators in the treatment of acute mercury intoxication
rats were studied. Male albino rats were divided in to various groups of six animals each. Group 1 served as control. Groups 2 and 3 were treated with NAC and DTT per se groups. Groups 4 and 6 were administered dimethylmercury (10 g/kg, p.o.) for once only; group 4 was served as experimental control. Groups 5 and 6 were treated with the conjoint treatment of NAC and DTT along with Zn and Se. Compared to the control, significant increase (P < 0.05) was observed in the activities of AST, ALT, SALP and LDH after toxicant administration. A significant rise was observed in LPO level however reduced GSH content in liver, kidney and brain. Drug metabolizing enzymatic activities (AH/AND) in liver microsomes was also decreased. A significant decrease was noted in the activity of AChE in different regions of brain Thus, the co-administration of zinc and selenium during chelation treatment with NAC and DTT could be recommended for achieving optimum effects of chelation therapy. Improvement in the ultra structure and histoarchitecture of liver, kidney and brain also supported the biochemical studies. doi:10.1016/j.toxlet.2010.03.690
P206-002 Antioxidant and hepatoprotective activity of polygonum bistorta (Linn.) and its active principle against acetaminophen-induced liver toxicity in male rats D.K. Mittal School of Studies in Zoology, Jiwaji University, Gwalior (M.P.), India Many herbal preparations have been recommended in alternative system of medicine for the treatment of hepatic disorder. Polygonum bistorta is powerful astringent, demulcent, diuretic, febrifuge, laxative, strongly styptic and rich in tannins. It is well known that overdoses of acetatminophen are hepatotoxic. Hepatoprotective activity of Polygonum bistorta and tannic acid was studied using acetaminophen -induced hepatotoxicity in rat model. Male Sprague–Dawley strain rats were administered a single bolus dose of acetaminophen (2 g/kg, p.o.). Plant extract (Polygonum bistorta) and its active principle (tannic acid) was given at the dose of 100 and 25 mg/kg, respectively (p.o.) after 24 h of toxicant administration. The hepatotoxicity produced by acute paracetamol administration was found to be inhibited by Polygonum bistorta and tannic acid with evidence of decreased levels of serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, protein, albumin, bilirubin, urea, creatinine, triglycerides and cholesterol concentration whereas increase was found in blood sugar. Histopathological findings also suggest that plant extract and its active principle prevent the development of acute liver damage. These changes was assigned to the study of DNA damage (COMET assay), thus supported these biochemical and histopathological findings. The results of this study clearly indicate that Polygonum bistorta and tannic acid have a potent hepatoprotective action; whereas tannic acid is showed more protective effect compare to polygonum bistorta against acetaminophen -induced toxicological profile. doi:10.1016/j.toxlet.2010.03.691
D. Joshi School of studies in Zoology, Jiwaji University, Gwalior, India Public awareness of the potential for mercury to cause health problems has increased dramatically in the last 15 years. Ameliorative effects of few naturally occurring antioxidants and chelating agents like NAC, DTT, zinc and selenium on parameters indicative of oxidative stress in the liver, kidney, brain and blood of mercury-exposed
Abstracts / Toxicology Letters 196S (2010) S37–S351
P205-037 Marmoset monkeys as preclinical models for respiratory diseases S. Seehase 1 , H.D. Lauenstein 1 , S. Switalla 1 , F. Prenzler 1 , F.J. Kaup 2 , E. Fuchs 2 , N. Krug 1 , C. Schlumbohm 2 , K. Sewald 1 , A. Braun 1 1
Fraunhofer Institute for Toxicology and Experimental Medicine, Germany, 2 German Primate Center, Germany Therapeutic efficacy studies of biopharmaceuticals for respiratory diseases are often limited in laboratory animals due to high interspecies differences. In order to adapt preclinical models closer to the human situation we are currently establishing lipopolysaccharide (LPS)-induced inflammation models in a non-human primate, the marmoset monkey (Callithrix jacchus). Whole blood (WB) samples were incubated with LPS in vitro to determine the individual immunological potential of the animals. Precision cut lung slices (PCLS) were used to investigate the immunological potential in the target organ lung. Dexamethasone served as treatment control in both, PCLS and WB-assay. In an in vivo approach marmosets were intrabronchially exposed to LPS. Before LPS instillation a unilateral bronchoalveolar lavage (BAL) was performed in anaesthetized animals under bronchoscopical supervision to determine BAL cell counts and TNF-alpha release at baseline levels. LPS was instilled in anesthetized animals intratracheally into the left lung via microsprayer. Thereafter a second BAL was performed 18 h later. Cell counts and TNF-alpha concentration were determined in BAL fluid. To monitor systemic effects, haematology, body weight and TNF-alpha concentration in serum were analyzed before and after LPS provocation. Both, WB-assay and PCLS showed a dose-dependent TNF-alpha response to LPS. Significant correlations were found between PCLS and WB-assay (r2 = 0.9). The same was true for human and marmoset PCLS (r2 = 0.9). Dexamathasone treatment reduced TNFalpha release significantly (WBA to 59%, PCLS to 77%). In the in vivo approach LPS instillation caused a moderate TNF-alpha increase (<5 pg/ml vs. 28 pg/ml) and a strong neutrophilic influx in BAL fluid (<1% vs. 79% after LPS), approved by histology and BAL cell counts. No signs of systemic inflammation were detectable. In conclusion, marmoset monkeys represent a suitable model to study inflammatory effects in the lung and their therapeutical intervention in vitro, ex vivo, and in vivo, close to the human situation. doi:10.1016/j.toxlet.2010.03.688
P206 Target Organ Toxicity. Hepatotoxicity P206-001 Beneficial effect of combined administration of some naturally occurring antioxidants and thiol chelators in the treatment of acute mercury intoxication
rats were studied. Male albino rats were divided in to various groups of six animals each. Group 1 served as control. Groups 2 and 3 were treated with NAC and DTT per se groups. Groups 4 and 6 were administered dimethylmercury (10 g/kg, p.o.) for once only; group 4 was served as experimental control. Groups 5 and 6 were treated with the conjoint treatment of NAC and DTT along with Zn and Se. Compared to the control, significant increase (P < 0.05) was observed in the activities of AST, ALT, SALP and LDH after toxicant administration. A significant rise was observed in LPO level however reduced GSH content in liver, kidney and brain. Drug metabolizing enzymatic activities (AH/AND) in liver microsomes was also decreased. A significant decrease was noted in the activity of AChE in different regions of brain Thus, the co-administration of zinc and selenium during chelation treatment with NAC and DTT could be recommended for achieving optimum effects of chelation therapy. Improvement in the ultra structure and histoarchitecture of liver, kidney and brain also supported the biochemical studies. doi:10.1016/j.toxlet.2010.03.690
P206-002 Antioxidant and hepatoprotective activity of polygonum bistorta (Linn.) and its active principle against acetaminophen-induced liver toxicity in male rats D.K. Mittal School of Studies in Zoology, Jiwaji University, Gwalior (M.P.), India Many herbal preparations have been recommended in alternative system of medicine for the treatment of hepatic disorder. Polygonum bistorta is powerful astringent, demulcent, diuretic, febrifuge, laxative, strongly styptic and rich in tannins. It is well known that overdoses of acetatminophen are hepatotoxic. Hepatoprotective activity of Polygonum bistorta and tannic acid was studied using acetaminophen -induced hepatotoxicity in rat model. Male Sprague–Dawley strain rats were administered a single bolus dose of acetaminophen (2 g/kg, p.o.). Plant extract (Polygonum bistorta) and its active principle (tannic acid) was given at the dose of 100 and 25 mg/kg, respectively (p.o.) after 24 h of toxicant administration. The hepatotoxicity produced by acute paracetamol administration was found to be inhibited by Polygonum bistorta and tannic acid with evidence of decreased levels of serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, protein, albumin, bilirubin, urea, creatinine, triglycerides and cholesterol concentration whereas increase was found in blood sugar. Histopathological findings also suggest that plant extract and its active principle prevent the development of acute liver damage. These changes was assigned to the study of DNA damage (COMET assay), thus supported these biochemical and histopathological findings. The results of this study clearly indicate that Polygonum bistorta and tannic acid have a potent hepatoprotective action; whereas tannic acid is showed more protective effect compare to polygonum bistorta against acetaminophen -induced toxicological profile. doi:10.1016/j.toxlet.2010.03.691
D. Joshi School of studies in Zoology, Jiwaji University, Gwalior, India Public awareness of the potential for mercury to cause health problems has increased dramatically in the last 15 years. Ameliorative effects of few naturally occurring antioxidants and chelating agents like NAC, DTT, zinc and selenium on parameters indicative of oxidative stress in the liver, kidney, brain and blood of mercury-exposed