- Email: [email protected]
Antiparkinson and antioxidant effect of pyridylpyrazolinyl substituted heterosteroids in LPS induced neuroinflammation model of neurodegenerative disorders
e186
Abstracts / Parkinsonism and Related Disorders 22 (2016) e149ee192
test motor activity and to influence the action of reserpine autonomic manifestations: ptosis, diarrhea, salivation, decrease in body temperature. Substance is administered 30 minutes after the administration of reserpine. Evaluation is estimated after the motor activity for 10 minutes after 2 hours after the administration of reserpine. Assessment of autonomic disorders. Ptosis recorded in points largest eye slit 3 - complete closure of eyes, 2 - the gap to 1 mm, 1 - a gap of 2 mm, 0 - eyes fully open. Salivation estimated by size of wet spots on the neck: 3 - 2 cm, 2 - 1 cm, 1 - 0.5 cm, 0 no effect. Also registries presence of diarrhea. Conclusions: It should be noted that this model has several drawbacks. However, these Methods expressed model of oligokinesia and allow us to establish the presence of dopamine level in the mechanism of action of the test compound and is therefore widely used for the evaluation of antiparkinsonian drug preparations. P 6.047. EXOGENOUS SALSOLINOL INFLUENCES GUT-BRAIN AXIS Magdalena Kurnik, Krzysztof Gil, Magdalena Bialas, Veronika Aleksandrovych, Andrzej Bugajski, Piotr Thor. Department of Pathophysiology, Jagiellonian University Medical College, Krakow, Poland Objectives: Salsolinol (1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) is an endogenous compound thought to be involved in the etiology of Parkinson's disease. In animal studies, chronic administration of salsolinol induced parkinsonian-like symptoms. However, little is known about its effects on the gut-brain axis activation. The aim of our study was to investigate the influence of salsolinol on body weight gain, adipose tissue accumulation as well as corticosterone and corticotropinreleasing factor (CRF) serum levels. Methods: Male Wistar rats were subjected to continuous intraperitoneal low dosing of salsolinol (200 mg/kg in total) with osmotic mini-pumps for two or four weeks (S1, S2). Appropriate groups served as the controls (C1, C2). Blood samples were collected at the end of the experiment and assayed by ELISA. Both epididymal fat pads were dissected, weighted and processed for routine histology and image analysis. Results: Salsolinol significantly reduced animals body weight gain however there were no differences in the cumulative food intake between the groups. The epididymal fat pad/body weight ratio was significantly lower in salsolinol-treated rats for four weeks (S2 ¼ 10.57‰ ± 0.6 vs. C2 ¼ 13.86‰ ± 1.47). Corticosterone level was significantly increased in salsolinol-treated rats after two weeks (S1 ¼ 34.29 ng/ml ±3.74 vs. C1 ¼ 16.11ng/ ml ± 10.78). CRF levels were decreased in salsolinol-treated animals. The area and the perimeter of the fad pad adipocytes were slightly decreased after salsolinol treatment in comparison to control groups. Conclusions: Exogenous salsolinol targets some neuroendocrine circuits and regulatory mechanisms concerned with adipose tissue accumulation. P 6.048. CHARACTERIZATION OF THE LIPOPOLYSACCHARIDE INDUCED MODEL OF PARKINSON'S DISEASE: ROLE OF OXIDATIVE STRESS AND NEUROINFLAMMATION
increase in the mRNA expression of proinflammatory cytokines i.e. TNF-a and IL-1b were observed after 7 days of LPS infusion whereas the alterations in the oxidative stress markers i.e ROS, Lipid peroxidation, NO formation, NADPH oxidase activity, Glutathione system, SOD and catalase became highly significant after 14 days of infusion. As a consequence after 21 days of LPS infusion we observed activation of apoptotic pathway indicated by increased expression of caspases 3 and caspase 9. This was followed by a significant decline in the expression of tyrosine hyroxylase (TH) as revealed by IHC. Further a marked decrease in the level of dopamine and its metabolites enough for the production of behavioral abnormality in rats. Conclusion: Hence, the present study provides extensive characterization of LPS induced model of PD. Study also confirms the co-existence and complex interplay between inflammation and oxidative stress contributing equally to the dopaminergic neuronal degeneration process in PD. P 6.049. ANTIPARKINSON AND ANTIOXIDANT EFFECT OF PYRIDYLPYRAZOLINYL SUBSTITUTED HETEROSTEROIDS IN LPS INDUCED NEUROINFLAMMATION MODEL OF NEURODEGENERATIVE DISORDERS Ranjit Singh, Ranju Bansal. University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh, India Objectives: Parkinson Disease (PD) is a progressing, disabling neurodegenerative disorder characterized by an insidious onset with variable expression of motor, vegetative, sensory and psychopathological symptoms. Recent literature reports indicated that neuroinflammation cause dopaminergic neuron death that involved in progression of PD. The study is aimed at design and synthesis of new therapeutically useful steroidal neuroprotective agents against neurodegenerative disorders. New androstane derivatives have been synthesized by fusing pyridylpyrazoline moiety at 17 and 16 position of steroid nucleus. These D ring modified heterosteroids were then explored for their antiparkinson and antioxidant effect. Methods: Aldol condensation of dehydroepiandrosterone with requisite pyridine carboxaldehyde in basic medium gave corresponding 16-arylidene steroidal derivatives, treatment of which with hydrazine hydrate in refluxing 1,4 dioxane afforded pyrazoline substituted steroids 1a-c. Rats (male wistar) were anesthetized with thiopental sodium (45 mg/Kg, i.p.), stereotaxic surgery has been done and LPS (10mg in 2ml) was infused into left substantia nigra using Hamilton microsyringe. Heterosteroids were than evaluated for behavioral alternation using actophotometer, elevated plus maze and wooden blocks at dose 2 mg/Kg (i.p) after 7th,14th and 21st day of LPS administration. Biochemical estimation of different markers for neuroinflammation, cholinergic activity and oxidative stress has also been carried out. Results: The synthetic heterosteroids were characterized using IR, H1NMR. All heterosteroids 1a-c exhibited potent activity using against PD. The pyrindin-4-yl group substituted steroid 1c displayed comparable to that of standard dexamethasone and celecoxib. Conclusions: Pyridylpyrazoline substituted heterosteroids exhibited potent anti-neuro-inflammatory activity and could be useful for the prevention of PD and oxidative stress.
Neha Sharma, Bimla Nehru. Department of Biophysics, Panjab University, Chandigarh, India Introduction: Primary pathology underlying Parkinson's disease (PD) is the loss of dopaminergic neurons in the Substantia Nigra (SN). A variety of genetic and environmental factors underlie this loss of dopaminergic neurons. However, recent studies have highlighted the role of elevated oxidative stress and the pro-inflammatory responses contributing to or exacerbating the nigrostriatal degeneration. Methods: With the establishment of neuroinflammation as an important process involved in the PD pathogenesis, in the present study this pathogenic feature was replicated in animals using lipopolysaccharide (LPS) (5ug/5ul PBS) infused stereotaxically into the SN of rats. Results: LPS injected into the SN successfully replicated the pathogenic features of PD in rats as it elicited an inflammatory response via action of microglia. LPS infusion resulted in glial cell activation as depicted from Immunohistochemistry (IHC) analysis of GFAP and Iba-1. Also, a significant Ă
Abstracts / Parkinsonism and Related Disorders 22 (2016) e149ee192
test motor activity and to influence the action of reserpine autonomic manifestations: ptosis, diarrhea, salivation, decrease in body temperature. Substance is administered 30 minutes after the administration of reserpine. Evaluation is estimated after the motor activity for 10 minutes after 2 hours after the administration of reserpine. Assessment of autonomic disorders. Ptosis recorded in points largest eye slit 3 - complete closure of eyes, 2 - the gap to 1 mm, 1 - a gap of 2 mm, 0 - eyes fully open. Salivation estimated by size of wet spots on the neck: 3 - 2 cm, 2 - 1 cm, 1 - 0.5 cm, 0 no effect. Also registries presence of diarrhea. Conclusions: It should be noted that this model has several drawbacks. However, these Methods expressed model of oligokinesia and allow us to establish the presence of dopamine level in the mechanism of action of the test compound and is therefore widely used for the evaluation of antiparkinsonian drug preparations. P 6.047. EXOGENOUS SALSOLINOL INFLUENCES GUT-BRAIN AXIS Magdalena Kurnik, Krzysztof Gil, Magdalena Bialas, Veronika Aleksandrovych, Andrzej Bugajski, Piotr Thor. Department of Pathophysiology, Jagiellonian University Medical College, Krakow, Poland Objectives: Salsolinol (1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline) is an endogenous compound thought to be involved in the etiology of Parkinson's disease. In animal studies, chronic administration of salsolinol induced parkinsonian-like symptoms. However, little is known about its effects on the gut-brain axis activation. The aim of our study was to investigate the influence of salsolinol on body weight gain, adipose tissue accumulation as well as corticosterone and corticotropinreleasing factor (CRF) serum levels. Methods: Male Wistar rats were subjected to continuous intraperitoneal low dosing of salsolinol (200 mg/kg in total) with osmotic mini-pumps for two or four weeks (S1, S2). Appropriate groups served as the controls (C1, C2). Blood samples were collected at the end of the experiment and assayed by ELISA. Both epididymal fat pads were dissected, weighted and processed for routine histology and image analysis. Results: Salsolinol significantly reduced animals body weight gain however there were no differences in the cumulative food intake between the groups. The epididymal fat pad/body weight ratio was significantly lower in salsolinol-treated rats for four weeks (S2 ¼ 10.57‰ ± 0.6 vs. C2 ¼ 13.86‰ ± 1.47). Corticosterone level was significantly increased in salsolinol-treated rats after two weeks (S1 ¼ 34.29 ng/ml ±3.74 vs. C1 ¼ 16.11ng/ ml ± 10.78). CRF levels were decreased in salsolinol-treated animals. The area and the perimeter of the fad pad adipocytes were slightly decreased after salsolinol treatment in comparison to control groups. Conclusions: Exogenous salsolinol targets some neuroendocrine circuits and regulatory mechanisms concerned with adipose tissue accumulation. P 6.048. CHARACTERIZATION OF THE LIPOPOLYSACCHARIDE INDUCED MODEL OF PARKINSON'S DISEASE: ROLE OF OXIDATIVE STRESS AND NEUROINFLAMMATION
increase in the mRNA expression of proinflammatory cytokines i.e. TNF-a and IL-1b were observed after 7 days of LPS infusion whereas the alterations in the oxidative stress markers i.e ROS, Lipid peroxidation, NO formation, NADPH oxidase activity, Glutathione system, SOD and catalase became highly significant after 14 days of infusion. As a consequence after 21 days of LPS infusion we observed activation of apoptotic pathway indicated by increased expression of caspases 3 and caspase 9. This was followed by a significant decline in the expression of tyrosine hyroxylase (TH) as revealed by IHC. Further a marked decrease in the level of dopamine and its metabolites enough for the production of behavioral abnormality in rats. Conclusion: Hence, the present study provides extensive characterization of LPS induced model of PD. Study also confirms the co-existence and complex interplay between inflammation and oxidative stress contributing equally to the dopaminergic neuronal degeneration process in PD. P 6.049. ANTIPARKINSON AND ANTIOXIDANT EFFECT OF PYRIDYLPYRAZOLINYL SUBSTITUTED HETEROSTEROIDS IN LPS INDUCED NEUROINFLAMMATION MODEL OF NEURODEGENERATIVE DISORDERS Ranjit Singh, Ranju Bansal. University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh, India Objectives: Parkinson Disease (PD) is a progressing, disabling neurodegenerative disorder characterized by an insidious onset with variable expression of motor, vegetative, sensory and psychopathological symptoms. Recent literature reports indicated that neuroinflammation cause dopaminergic neuron death that involved in progression of PD. The study is aimed at design and synthesis of new therapeutically useful steroidal neuroprotective agents against neurodegenerative disorders. New androstane derivatives have been synthesized by fusing pyridylpyrazoline moiety at 17 and 16 position of steroid nucleus. These D ring modified heterosteroids were then explored for their antiparkinson and antioxidant effect. Methods: Aldol condensation of dehydroepiandrosterone with requisite pyridine carboxaldehyde in basic medium gave corresponding 16-arylidene steroidal derivatives, treatment of which with hydrazine hydrate in refluxing 1,4 dioxane afforded pyrazoline substituted steroids 1a-c. Rats (male wistar) were anesthetized with thiopental sodium (45 mg/Kg, i.p.), stereotaxic surgery has been done and LPS (10mg in 2ml) was infused into left substantia nigra using Hamilton microsyringe. Heterosteroids were than evaluated for behavioral alternation using actophotometer, elevated plus maze and wooden blocks at dose 2 mg/Kg (i.p) after 7th,14th and 21st day of LPS administration. Biochemical estimation of different markers for neuroinflammation, cholinergic activity and oxidative stress has also been carried out. Results: The synthetic heterosteroids were characterized using IR, H1NMR. All heterosteroids 1a-c exhibited potent activity using against PD. The pyrindin-4-yl group substituted steroid 1c displayed comparable to that of standard dexamethasone and celecoxib. Conclusions: Pyridylpyrazoline substituted heterosteroids exhibited potent anti-neuro-inflammatory activity and could be useful for the prevention of PD and oxidative stress.
Neha Sharma, Bimla Nehru. Department of Biophysics, Panjab University, Chandigarh, India Introduction: Primary pathology underlying Parkinson's disease (PD) is the loss of dopaminergic neurons in the Substantia Nigra (SN). A variety of genetic and environmental factors underlie this loss of dopaminergic neurons. However, recent studies have highlighted the role of elevated oxidative stress and the pro-inflammatory responses contributing to or exacerbating the nigrostriatal degeneration. Methods: With the establishment of neuroinflammation as an important process involved in the PD pathogenesis, in the present study this pathogenic feature was replicated in animals using lipopolysaccharide (LPS) (5ug/5ul PBS) infused stereotaxically into the SN of rats. Results: LPS injected into the SN successfully replicated the pathogenic features of PD in rats as it elicited an inflammatory response via action of microglia. LPS infusion resulted in glial cell activation as depicted from Immunohistochemistry (IHC) analysis of GFAP and Iba-1. Also, a significant Ă