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Intrathecal CGRP 8-37 attenuates formalin-induced pain behavior and spinal cord fos expression in rats
Abstracts
Conclusion: We know the bone is composed of a protein matrix and the incorporation of the mineral is essential for the hardness and rigidity that enables the skeleton to resist gravitational and mechanical loading. Acid extraction removes the mineral thus the protein surface of the scaffold (flexible microenvironment) will be presented which is familiar for the cells. Keywords: MBA, DBM, Scaffold, 3D-culture, Mesenchymal progenitor cells doi:10.1016/j.clinbiochem.2011.08.139
E Poster – [A-10-908-2] Intrathecal CGRP 8-37 attenuates formalin-induced pain behavior and spinal cord fos expression in rats Takhshid Mohammad Ali, Owji Ali Akbar, Khoshdel Zahra Shiraz University of Medical Sciences, Shiraz, Iran E-mail addresses: [email protected] (T.M. Ali), [email protected] (O.A. Akbar), [email protected] (K. Zahra) Introduction: Calcitonin gene-related peptide (CGRP) has been proposed as a nociceptive transmitter in the dorsal horn of the spinal cord. The aim of this study was to examine potential antinociceptive effect of CGRP 8-37, a specific CGRP1 receptor antagonist, on formalin-induced pain behavior and its associated spinal cord fos expression in rats. Methods: Noxious chemical (formalin 5%, 50 μl) stimuli were applied to a hind paw of Sprague Dawley rats 15 minutes after intrathecal CGRP 8-37 (20 μg/rat) administration to study its antinociceptive effect. The expression c-fos was detected by immunohistochemistry. Results: Subcutaneous injection of formalin produced a marked biphasic pain response (first phase: 0–5 min and second phase: 15– 45 min). Compared with the physiological saline control, spinally administered CGRP 8-37 significantly decreased the first and late phase of formalin-induced pain. In addition, CGRP 8-37 significantly reduced formalin-induced c-fos expression in the L4–5 spinal dorsal horn neurons. Conclusion: These data suggest that CGRP 8-37 can exert antinociceptive effects in the formalin test by preventing neuronal activation at the spinal level, and CGRP1 receptors may involve in the spinal modulation of persistent nociception. Keywords: CGRP receptor, CGRP 8-37, Formalin test, C-fos expression doi:10.1016/j.clinbiochem.2011.08.140
E Poster – [A-10-1181-1] Survival of chicken somitic cells following co-culture with notochord Rahbari Rezgar, Sagha Mohsen, Mazani Mohammad Ardbil University of Medical Sciences, Ardbil, Iran E-mail address: [email protected] (R. Rezgar) Introduction: Somites are segmental blocks of paraxial mesoderm, located on either sides of developing neural tube and notochord. They can differentiate to bones, cartilages and muscles. The notochord, as a rod-shape mesoderm is located beneath the developing neural tube and plays a key role in somitic cell survival and differentiation. It is well understood that these biological activities of the notochord is due to sonic hedge hog (SHH) secretion. The aim of this study is to evaluate the in vitro survival effect of notochord on somites.
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Materials and methods: Upon isolation of chick embryonic somites and notochord, the notochord was encapsulated in alginate solution (1.2%). The somites were co-cultured with beads containing notochord for 4, 6 and 10 days in DMEM/F12 supplemented with 10% FBS. In control group, the somites were remained untreated and plated at the same days. Then, survival rate of somitic cells was assayed by MTT test. Results: Our results showed that 75% of somitic cells could survive upon 4 day co-culturing with notochord, compared to 27.5% in control group (p < 0.02). After 6 days, somitic cell survival raised to 87.5% which found to be significantly higher than control group (52.5%, p < 0.05). During 10 day co-culture, survival rate of somitic cells significantly declined to 35.7%. Conclusion: Our findings clearly showed that the notochord can mimic its survival-inducing ability in vitro and supports somitic cell survival after 4–6 days co-culturing. Keywords: Somitic cells, Notochord, Co-culture, Survival rate doi:10.1016/j.clinbiochem.2011.08.141
E Poster – [A-10-1220-1] Evaluation of 11-hydroxy-Δ8-THC-dimethylheptyl effects on cytokine profile and locomotor tests in an experimental model of multiple sclerosis Mohammad Esmaeil Shahaboddin, Mohammad Hossein Aarabi, Karim Parastouei, Mitra Motallebi, Akbar Jafarnejad, Mehdi Mirhashemi, Gholam Ali Hamidi Biochemistry Department, School of Medicine, Kashan University of Medical Sciences, Kashan, Iran E-mail address: [email protected] (M.E. Shahaboddin) Introduction: Multiple sclerosis is an inflammatory chronic demyelinating disease of the central nervous system. Most of the current therapeutic treatments for MS involve strategies related to immune modulation. In this line, several recent reports described the beneficial effects of cannabinoids, based on their anti-inflammatory and immunomodulatory properties. One of the synthetic components that activate cannabinoids receptors is 11-hydroxy-Δ8-THC-dimethylheptyl (hu-210). The present study was conducted to evaluate the effectiveness of hu-210 treatment to cytokine profile and clinical signs in the experimental autoimmune encephalomyelitis (EAE) model in mice. Materials and methods: In this experimental interventional study, totally 48 male C57BL/6 mice were placed in 8 therapeutic groups (n = 6 per group). Mice were immunized subcutaneously with 250 μg myelin oligodendrocyte glycoprotein (MOG) 35–55. When EAE was observed, HU210 was subcutaneously administered in 3 groups with different doses: 3, 10 and 30 mg/kg. Control groups (EAE and non-EAE) were treated to vehicle or HU210. Drug or vehicle was given once on the following days: 1, 3, 5, 7, 9, 11, 13, 15 and 17. In the study duration and clinical score of disease was analyzed. After 17 days of treatment, the animals were sacrificed and, serum TNFα, IL-12 levels as inflammatory factors, and serum IL-4 level as an anti-inflammatory factor were measured by ELISA assay. Results: Our results showed that HU210-treated mice had significantly less clinical score of EAE than non-treated EAE induced mice (p< 0.05). Our data revealed that a dose of 30 mg/kg HU210 in EAE induced mice significantly decreased serum TNFα, IL-12 (p< 0.05), but in low doses this reduction wasn't significant (p> 0.05). Also, Serum IL-4 levels increased significantly in mice using 3,10 and 30 mg/kg doses of HU210 (p< 0.05). Conclusion: In conclusion, the present study revealed the beneficial effect of HU 210 in the model of EAE affecting the stages of the
Conclusion: We know the bone is composed of a protein matrix and the incorporation of the mineral is essential for the hardness and rigidity that enables the skeleton to resist gravitational and mechanical loading. Acid extraction removes the mineral thus the protein surface of the scaffold (flexible microenvironment) will be presented which is familiar for the cells. Keywords: MBA, DBM, Scaffold, 3D-culture, Mesenchymal progenitor cells doi:10.1016/j.clinbiochem.2011.08.139
E Poster – [A-10-908-2] Intrathecal CGRP 8-37 attenuates formalin-induced pain behavior and spinal cord fos expression in rats Takhshid Mohammad Ali, Owji Ali Akbar, Khoshdel Zahra Shiraz University of Medical Sciences, Shiraz, Iran E-mail addresses: [email protected] (T.M. Ali), [email protected] (O.A. Akbar), [email protected] (K. Zahra) Introduction: Calcitonin gene-related peptide (CGRP) has been proposed as a nociceptive transmitter in the dorsal horn of the spinal cord. The aim of this study was to examine potential antinociceptive effect of CGRP 8-37, a specific CGRP1 receptor antagonist, on formalin-induced pain behavior and its associated spinal cord fos expression in rats. Methods: Noxious chemical (formalin 5%, 50 μl) stimuli were applied to a hind paw of Sprague Dawley rats 15 minutes after intrathecal CGRP 8-37 (20 μg/rat) administration to study its antinociceptive effect. The expression c-fos was detected by immunohistochemistry. Results: Subcutaneous injection of formalin produced a marked biphasic pain response (first phase: 0–5 min and second phase: 15– 45 min). Compared with the physiological saline control, spinally administered CGRP 8-37 significantly decreased the first and late phase of formalin-induced pain. In addition, CGRP 8-37 significantly reduced formalin-induced c-fos expression in the L4–5 spinal dorsal horn neurons. Conclusion: These data suggest that CGRP 8-37 can exert antinociceptive effects in the formalin test by preventing neuronal activation at the spinal level, and CGRP1 receptors may involve in the spinal modulation of persistent nociception. Keywords: CGRP receptor, CGRP 8-37, Formalin test, C-fos expression doi:10.1016/j.clinbiochem.2011.08.140
E Poster – [A-10-1181-1] Survival of chicken somitic cells following co-culture with notochord Rahbari Rezgar, Sagha Mohsen, Mazani Mohammad Ardbil University of Medical Sciences, Ardbil, Iran E-mail address: [email protected] (R. Rezgar) Introduction: Somites are segmental blocks of paraxial mesoderm, located on either sides of developing neural tube and notochord. They can differentiate to bones, cartilages and muscles. The notochord, as a rod-shape mesoderm is located beneath the developing neural tube and plays a key role in somitic cell survival and differentiation. It is well understood that these biological activities of the notochord is due to sonic hedge hog (SHH) secretion. The aim of this study is to evaluate the in vitro survival effect of notochord on somites.
S67
Materials and methods: Upon isolation of chick embryonic somites and notochord, the notochord was encapsulated in alginate solution (1.2%). The somites were co-cultured with beads containing notochord for 4, 6 and 10 days in DMEM/F12 supplemented with 10% FBS. In control group, the somites were remained untreated and plated at the same days. Then, survival rate of somitic cells was assayed by MTT test. Results: Our results showed that 75% of somitic cells could survive upon 4 day co-culturing with notochord, compared to 27.5% in control group (p < 0.02). After 6 days, somitic cell survival raised to 87.5% which found to be significantly higher than control group (52.5%, p < 0.05). During 10 day co-culture, survival rate of somitic cells significantly declined to 35.7%. Conclusion: Our findings clearly showed that the notochord can mimic its survival-inducing ability in vitro and supports somitic cell survival after 4–6 days co-culturing. Keywords: Somitic cells, Notochord, Co-culture, Survival rate doi:10.1016/j.clinbiochem.2011.08.141
E Poster – [A-10-1220-1] Evaluation of 11-hydroxy-Δ8-THC-dimethylheptyl effects on cytokine profile and locomotor tests in an experimental model of multiple sclerosis Mohammad Esmaeil Shahaboddin, Mohammad Hossein Aarabi, Karim Parastouei, Mitra Motallebi, Akbar Jafarnejad, Mehdi Mirhashemi, Gholam Ali Hamidi Biochemistry Department, School of Medicine, Kashan University of Medical Sciences, Kashan, Iran E-mail address: [email protected] (M.E. Shahaboddin) Introduction: Multiple sclerosis is an inflammatory chronic demyelinating disease of the central nervous system. Most of the current therapeutic treatments for MS involve strategies related to immune modulation. In this line, several recent reports described the beneficial effects of cannabinoids, based on their anti-inflammatory and immunomodulatory properties. One of the synthetic components that activate cannabinoids receptors is 11-hydroxy-Δ8-THC-dimethylheptyl (hu-210). The present study was conducted to evaluate the effectiveness of hu-210 treatment to cytokine profile and clinical signs in the experimental autoimmune encephalomyelitis (EAE) model in mice. Materials and methods: In this experimental interventional study, totally 48 male C57BL/6 mice were placed in 8 therapeutic groups (n = 6 per group). Mice were immunized subcutaneously with 250 μg myelin oligodendrocyte glycoprotein (MOG) 35–55. When EAE was observed, HU210 was subcutaneously administered in 3 groups with different doses: 3, 10 and 30 mg/kg. Control groups (EAE and non-EAE) were treated to vehicle or HU210. Drug or vehicle was given once on the following days: 1, 3, 5, 7, 9, 11, 13, 15 and 17. In the study duration and clinical score of disease was analyzed. After 17 days of treatment, the animals were sacrificed and, serum TNFα, IL-12 levels as inflammatory factors, and serum IL-4 level as an anti-inflammatory factor were measured by ELISA assay. Results: Our results showed that HU210-treated mice had significantly less clinical score of EAE than non-treated EAE induced mice (p< 0.05). Our data revealed that a dose of 30 mg/kg HU210 in EAE induced mice significantly decreased serum TNFα, IL-12 (p< 0.05), but in low doses this reduction wasn't significant (p> 0.05). Also, Serum IL-4 levels increased significantly in mice using 3,10 and 30 mg/kg doses of HU210 (p< 0.05). Conclusion: In conclusion, the present study revealed the beneficial effect of HU 210 in the model of EAE affecting the stages of the