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Measurement of SOD and catalase activities were carried out in control and WPS-exposed animals using spectrophotometric method with commercially available kits (Cayman Chemical). Measurement of TNF�� and IL-6 in lung tissue At the end of the exposure period to either WPS or air, animals were sacrificed by an overdose of sodium pentobarbital, and their lungs were quickly collected and rinsed with ice-cold PBS (pH 7.4) before homogenization in 50?mmol/L Tris buffer pH 7.4 containing 400?mmol/L NaCl and 0.5% Triton X-100 at 4��C. The homogenates [http://www.selleckchem.com/products/GDC-0941.html Pictilisib price] were centrifuged for 10?min at 3000?��?g to remove cellular debris, and the supernatants were used for further analysis (Nemmar et?al. 2013c,d). Protein content was measured by Bradford's method as described before. The concentrations of IL-6 and TNF �� in the lung were determined using ELISA Kits (Duo Set; R & D systems, Minneapolis, MN). Airway reactivity to methacholine In separate group of mice (n?=?8 in each group), airway hyperreactivity responses were measured using [https://en.wikipedia.org/wiki/Azastene Azastene] a forced oscillation technique (FlexiVent; SCIREQ). Airway resistance (R) was assessed after increasing exposures to methacholine. Mice were anesthetized with an intraperitoneal injection of pentobarbital (70?mg/kg). The trachea was exposed and an 18-gauge metal needle was inserted into the trachea. Mice were connected to a computer-controlled small animal ventilator and quasi-sinusoidally ventilated with a tidal volume of 10?mL/kg at a frequency of 150 breaths/min and a positive end-expiratory pressure of 2?cm H2O to achieve a mean lung volume close to that observed during spontaneous breathing. After measurement of a baseline, each mouse was challenged with methacholine aerosol, generated with an in-line nebulizer and administered directly through the ventilator for 5?sec, with increasing concentrations (0, 0.625, 1.25, 2.5, and 5?mg/mL). R was measured using a ��snapshot�� protocol each 20?sec for 2?min. The mean of these five values was used for each methacholine concentration, unless the coefficient of determination of a measurement was smaller than 0.95. For each mouse, R was plotted [http://www.selleckchem.com/products/ve-821.html selleck inhibitor] against methacholine concentration (from 0 to 5?mg/mL; Nemmar et?al. 2012b; Nemmar et?al. 2013a). Statistics All statistical analyses were performed with GraphPad Prism Software version 5 (San Diego, CA, USA). To determine whether parameters were normally distributed, the KS normality test was applied. Normally distributed data were analyzed using the unpaired t-test for differences between groups. Nonnormally distributed data (total proteins and LDH in BAL fluid and ROS in lung) were analyzed using Mann�CWhitney test for differences between groups. All the data in figures were reported as mean?��?SEM. P values?

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Cloth59butter : Page créée avec « Measurement of SOD and catalase activities were carried out in control and WPS-exposed animals using spectrophotometric method with commercially available kits (Cayman Che... »