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Methods Experimental Protocol

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The experimental protocols were approved by the Institutional Animal Care and Use Committee of Harbin Medical University, and conducted in compliance with the animal-use guidelines (SYXK (Hei) 2006–033). Seventy-two adult Wistar rats (weight 250–300 g) were anesthetized with an intraperitoneal injection of 30 mg/kg of pentobarbital sodium. The internal carotid artery was cannulated with a 20-gauge catheter to aspirate blood for blood gas analysis and arterial pressure monitoring. The rectal temperature was maintained at 37.0–38.5°C. Mechanical ventilation delivered via tracheostomy was initiated in the pressure-controlled mode (Kent Scientific Ventilator-Dual Mode, USA) with 15 cmH2O PIP, a positive end-expiratory pressure (PEEP) of 2 cmH2O, a frequency of 30 breaths/min to maintain the PaCO2 at 35–45 mmHg, an inspiration-to-expiration (I:E) ratio of 1:2, and an inhaled oxygen fraction (FiO2) of 0.7 for 15 min, after which baseline data were collected.

Prior to randomization, the following values needed to be stable: PaO2/FiO2 > 300 mmHg, PaCO2 30–45 mmHg, and HCO3− > 20 mmol · L−1. If any parameter was not fulfilled, the animals were excluded from the protocol and further data analysis.

Experimental Groups

Seventy two rats were randomly assigned to 8 blocks of 9 animals each, with random numbers generated by SPSS (version 13.01S; Beijing Stats Data Mining Co. Ltd, Beijing, China). Among them, two blocks were randomly assigned to the sham group (anaesthetized and non-ventilated rats) and NV group (ventilated with PIP = 15 cmH2O and inhaled FiO2 of 0.7 for 4 h) served as controls for assessing the expression of NF-κB p65 protein and the inflammatory mediators in the lung. The left 6 blocks were assigned to three groups through merging two blocks of rats randomly, and including the Normocapnia (NC) group (PaCO2 = 35–45 mmHg, n = 18), PaCO2 was maintained in the normal range through inhaling the gas mixture (FiO2 0.7, FiCO2 4–5%, balance N2); the Moderate Hypercapnic Acidosis (MHA) group (PaCO2 = 80–100 mmHg, n = 18), PaCO2 was maintained through inhaling the gas mixture (FiO2 0.7, FiCO2 11–12%, balance N2); and the Severe Hypercapnic Acidosis (SHA) group (PaCO2 = 130–150 mmHg, n = 18), PaCO2 was maintained through inhaling the gas mixture (FiO2 0.7, FiCO2 16–17%, balance N2). The rats in the three groups were ventilated for 4 h in the supine position with a PIP of 30 cmH2O via the pressure-controlled mode (inspiratory time = 0.7 s; PEEP = 2 cmH2O and respiratory rate = 30 breaths/min). For all rats, anesthesia was maintained with sodium pentobarbital (2–4 mg · kg−1 · hr−1) and pancuronium bromide (0.03–0.07 mg · kg−1 · hr−1). Throughout the experiment, frequent checks were made to ensure that the animals were adequately anesthetized. This was performed by applying a painful stimulus to a paw and observing blood pressure responses. Lactated Ringer’s solution was infused i.V. At 10 ml · kg−1 · hr−1 to compensate for blood sampling.

 

 

 

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