Fourteen days later, mice were challenged with an intrathoracic injection of egg albumin (50 L, 12.5 g/cavity).[26,27] Briefly, an adapted needle was inserted into the right side of the thoracic cavity of egg albumin sensitized animals to permit the intrapleural administration of egg albumin diluted in sterile pyrogen free saline (50 L). The pathophysiology of asthma involves multiple factors leading to hyper-responsiveness and bronchoconstriction, such as mast cell degranulation,[12] neurogenic dysfunction,[13] involvement of T-lymphocytes and eosinophils,[14,15] altered immunosuppressive macrophages,[16,17] overproduction of proinflammatory cytokines,[18,19] immunoglobulins (IgE, IgG, and IgD)[20] and plasma protein exudation.[21C23] Therefore, the present work aims to prove the antiallergic activity based on studies focusing upon inflammation of the airway. MATERIALS AND METHODS Plant Aerial parts of were obtained from commercial supplier of Ahmedabad. The plant was identified and authenticated by Dr. Minoo Parabia, Head and Professor, Department of Bioscience, Veer Narmad South Gujarat University, Surat, Gujarat. A voucher specimen of plant was deposited in the herbarium of the Department of Bioscience, Veer Narmad South Gujarat University, Surat, Gujarat. Preparation of the plant extract The aerial parts reduced to coarse powder were macerated with ethanol for 48 h, filtered, and filtrate was evaporated under reduced pressure to obtain a dry extract. The ethanolic extract of (referred to as EtMe) was stored in cool and dry place and used for pharmacological evaluation. Reagents Egg albumin and aluminum hydroxide hydrate gel (alum) were purchased from S. D. Fine Chem Limited, India. Freunds complete adjuvant emulsion, and Evans blue were purchased from HiMedia, India. All other chemicals used were of analytical grade. Rabbit Polyclonal to Cytochrome P450 2A7 Animal protocol Swiss albino mice of either sex housed in standard conditions of temperature (22 2C), relative humidity (55 5%), and light (12 hrs light/dark cycles) were used. Animal studies were approved by the Institutional Animal Ethics Committee (protocol no 8004), as per the guidelines of committee for the purpose of control and supervision of experiments on animals (CPCSEA). Vascular permeability induced by acetic acid[24,25] Swiss albino mice were randomly allocated to four groups each containing six animals. Group I (model control), Group II (standard), Group III (EtMe), and Group IV (EtMe) received saline, indomethacin (20 mg/kg), EtMe (75 mg/kg), and EtMe (150 mg/kg) orally, respectively. Thirty minutes later, mice received an intravenous injection of 0.5% Evans blue saline solution (0.1 ml/10 g body weight) and an intraperitoneal injection of 0.6% acetic acid (10 ml/kg). After 20 min, the dye that leaked into the peritoneal cavity was collected by lavaging with 10 ml distilled water and was transferred to 10 ml volumetric flask through glass wool. To each flask, 0.1 ml of 0.1 N sodium hydroxide solution was added and volume made upto the mark with distilled water followed by measurement of absorbance at 610 nm (Shimadzu Spectrophotometer). Allergic pleurisy Swiss albino mice were randomly allocated to five groups each containing six animals: Group I (normal control), Group II (model control), Group III (standard), Group IV (EtMe), and Group V (EtMe). Active sensitization of Swiss albino mice was achieved with a subcutaneous injection of Freunds complete adjuvant emulsion (100 L) containing egg albumin (100 g). Fourteen days later, mice were challenged with an intrathoracic injection of egg albumin (50 L, 12.5 g/cavity).[26,27] Briefly, an adapted needle was inserted into the right side of the thoracic cavity of egg albumin sensitized animals to permit the intrapleural administration of egg albumin diluted in sterile pyrogen free saline (50 L). At 24 h after the stimulus, mice were anesthetized and their thoracic cavities were rinsed with 1 mL NS6180 phosphate buffer saline containing 10 mM EDTA, pH 7.4. Total leukocyte counts were made with an automated cell counter (Cell Dyn 3200SL). Differential cell counts were made by light microscopy stained with Leishmans stain. Groups IV and V animals fasted.The oral pretreatment with EtMe (75 mg/kg and 150 mg/ kg, po) markedly inhibited the eosinophil accumulation as compared to the control group and this inhibition was selective for decrease in the number of eosinophils. cytology in guinea pigs.[10,11] The pathophysiology of asthma involves multiple factors leading to hyper-responsiveness and bronchoconstriction, such as mast cell degranulation,[12] neurogenic dysfunction,[13] involvement of T-lymphocytes and eosinophils,[14,15] altered immunosuppressive macrophages,[16,17] overproduction of proinflammatory cytokines,[18,19] immunoglobulins (IgE, IgG, and IgD)[20] and plasma protein exudation.[21C23] Therefore, the present work aims to prove the antiallergic activity based on studies focusing upon inflammation of the airway. MATERIALS AND METHODS Plant Aerial parts of were obtained from commercial supplier of Ahmedabad. The plant was identified and authenticated by Dr. Minoo Parabia, Head and Professor, Department of Bioscience, Veer Narmad South Gujarat University, Surat, Gujarat. A voucher specimen of plant was deposited in the herbarium of the Department of Bioscience, Veer Narmad South Gujarat University, Surat, Gujarat. Preparation of the plant extract The aerial parts reduced to coarse powder were macerated with ethanol for 48 h, filtered, and filtrate was evaporated under reduced pressure to obtain a dry extract. NS6180 The ethanolic extract of (referred to as EtMe) was stored in cool and dry place and used for pharmacological evaluation. Reagents Egg albumin and aluminum hydroxide hydrate gel (alum) were purchased from S. D. Fine Chem Limited, India. Freunds complete adjuvant emulsion, and Evans blue were purchased from HiMedia, India. All other chemicals used were of analytical grade. Animal protocol Swiss albino mice of either sex housed in standard conditions of temperature (22 2C), relative humidity (55 5%), and light (12 hrs light/dark cycles) were used. Animal NS6180 studies were approved by the Institutional Animal Ethics Committee (protocol no 8004), as per the guidelines of committee for the purpose of control and supervision of experiments on animals (CPCSEA). Vascular permeability induced by acetic acid[24,25] Swiss albino mice were randomly allocated to four groups each containing six animals. Group I (model control), Group II (standard), Group III (EtMe), and Group IV (EtMe) received saline, indomethacin (20 mg/kg), EtMe (75 mg/kg), and EtMe (150 mg/kg) orally, respectively. Thirty minutes later, mice received an intravenous injection of 0.5% Evans blue saline solution (0.1 ml/10 g body weight) and an intraperitoneal injection of 0.6% acetic acid (10 ml/kg). After 20 min, the dye that leaked into the peritoneal cavity was collected by lavaging with 10 ml distilled water and was transferred to 10 ml volumetric flask through glass wool. To each flask, 0.1 ml of 0.1 N sodium hydroxide solution was added and volume made upto the mark with distilled water followed by measurement of absorbance at 610 nm (Shimadzu Spectrophotometer). Allergic pleurisy Swiss albino mice were randomly allocated to five groups each containing six animals: Group I (normal control), Group II (model control), Group III (standard), Group IV (EtMe), and Group V (EtMe). Active sensitization of Swiss albino mice was achieved with a subcutaneous injection of Freunds complete adjuvant emulsion (100 L) containing egg albumin (100 g). Fourteen days later, mice were challenged with an intrathoracic injection of egg albumin (50 L, 12.5 g/cavity).[26,27] Briefly, an adapted needle was inserted into the right side of the thoracic cavity of egg albumin sensitized animals to permit the intrapleural administration of egg albumin diluted in sterile pyrogen free saline (50 L). At 24 h after the stimulus, mice were anesthetized and their thoracic cavities were rinsed with 1 mL phosphate buffer saline containing 10 mM EDTA, pH 7.4. Total leukocyte counts were made with an automated cell counter (Cell Dyn 3200SL). Differential cell counts were made by light microscopy stained with Leishmans stain. Groups IV and V animals fasted overnight and received EtMe (75 mg/kg) and EtMe (150 mg/kg) orally 1 h before stimulation. Group II animals were similarly treated with vehicle alone. In Group III, dexamethasone was given intraperitoneally (2 mg/kg) 24 and 1 h before stimulation. Statistical analysis The results of various studies were expressed as mean SEM and analyzed statistically using one-way ANOVA with Dunnett test to find out the level of significance. Data were considered statistically significant at P 0.05. RESULTS Effect of EtMe on vascular permeability induced by acetic acid Pretreatment with EtMe (75 mg/kg) and EtMe (150 mg/ kg) significantly inhibited the rise in.