LPS was purchased from Sigma (Escherichia coli 055:B5, USA). IL-1β and TNF-α were from Peprotech (USA). Evans blue, the NF-κB inhibitor PDTC, the p38 inhibitor SB203580, the ERK1/2 inhibitor U0126, and the p300 inhibitor curcumin were from Sigma (USA). The JNK inhibitor SP600125 was from Cell Signaling Technology (USA). A fluorescent dye calcein-AM used for live cell imaging was from Invitrogen (USA). The antibodies against p65, p38, phospho-p38, p44/42 MAPK, phospho-p44/42 MAPK, JNK, phospho-JNK, GFAP, and NF-κB were from Cell Signaling Technology (USA). The AQP4 antibodies were from Chemicon (USA). CRH was obtained from Tocris (1151, UK), and CRHR1 antagonist, CP154,526, was kindly donated by the Pfizer company (USA). The complementary DNA (cDNA) sequences encoding rat AQP4 (NM_001142366) were synthesized (Sangon Biotech, Shanghai, China) and then inserted into pmRFP-C1 at the XhoI and EcoRI sites.
Cp154 526
Manufactured by Pfizer
Sourced in United Kingdom, United States
CP154,526 is a laboratory equipment product designed for use in scientific research and analysis. It serves as a tool for various experimental and measurement purposes, but a detailed description of its core function cannot be provided while maintaining an unbiased and factual approach. More information may be available from the manufacturer.
Automatically generated - may contain errors
Lab products found in correlation
Calcein am, by Thermo Fisher Scientific (1 mentions)
Il 1β, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Evans blue, by Merck Group (1 mentions)
Curcumin, by Merck Group (1 mentions)
Sb203580, by Merck Group (1 mentions)
U0126, by Merck Group (1 mentions)
Phospho p44 42 mapk, by Cell Signaling Technology (1 mentions)
Glial fibrillary acidic protein (gfap), by Cell Signaling Technology (1 mentions)
Phospho jnk, by Cell Signaling Technology (1 mentions)
Tnf α, by Thermo Fisher Scientific (1 mentions)
Sp600125, by Cell Signaling Technology (1 mentions)
P44 42 mapk, by Cell Signaling Technology (1 mentions)
Nf κb, by Cell Signaling Technology (1 mentions)
Protease inhibitor, by Roche (1 mentions)
Tween 80, by Merck Group (1 mentions)
Nickel sulfate, by Merck Group (1 mentions)
Avidin biotin complex, by Vector Laboratories (1 mentions)
4 protocols using cp154 526
1
Inhibition of Inflammatory Pathways
2
Morphine Hydrochloride Administration Protocol
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Morphine hydrochloride was obtained from Alcaliber Labs (Madrid, Spain), dissolved in sterile 0.9% saline and injected interperitoneally (i.p.) in a volume of 0.1 ml/10 g of body weight. Reagents used were: protease inhibitors (Roche Diagnostics, Indianapolis, IN); phosphatase inhibitor cocktail set (Calbiochem, San Diego, CA); goat and horse serum (Sigma-Aldrich); avidin-biotin complex (Vector Laboratories, Burlingame, CA); and nickel sulfate (Sigma-Aldrich). CP-154,526, kindly provided by Pfizer (New York, NY), was dissolved in 10% Tween 80 (Sigma-Aldrich).
3
Hypoxia-Induced Physiological Changes in Rats
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Rats in the hypoxia group were placed in a transparent and ventilated hypobaric chamber (Avic Guizhou Fenglei Aviation Armament Co., Ltd, China, FLYDWC-50-IIC) and exposed to hypobaric hypoxia of 5000 m (10.8% O2) for, 1 day, or 5 days (21 (link)). To avoid isolation stress, groups of 6 rats were kept in one cage under hypoxia; the average amount of group food and water intake (g or ml/rat/day) was measured after hypoxia. The normoxia group was placed in an identical chamber set at sea level (20.9% O2). In validation experiments, rats received injections of the CRHR1 antagonist CP-154,526 or saline (30 mg/kg, ip, donated by Pfizer, Groton, CT) (19 (link), 21 (link)) before hypoxic stress (Figure 5A
4
Hypoxia Preconditioning in Rat Model
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A short period hypoxia (SH). Healthy adult male Sprague-Dawley rats (Experimental Animal Center, Zhejiang,
China) weighing 180 ± 20 g were group-housed in the behavior lab 7 days for environmental adaptation before experiments. Rats in the hypoxia group were placed in a hypobaric chamber and exposed to hypobaric hypoxia of 7000 m altitude (~8.2% O 2 )for 1, 8, 24 h (Fig. 2A, B,C,F,G,H,J) or a prolonged hypoxia (PH), which mimicked at altitude of 5000 m (~10.8% O 2 ) for 2 or 5 days (4 h/per day, Fig. 2D,E,I) (Hao et al., 2015) . The normoxia group (Control, Con) was placed in the same chamber set at sea level (~ 21 % O 2 ). Rats were randomized into different groups. 1. The Control group was injected (ip) with 0.9% saline. 2. The Hypoxia group was injected with vehicle (0.9% saline) before hypoxia stress. 3. Dex group was injected with Dexamethasone (Dex, 500 µg/kg, ip) for 2 days (4 h hypoxia /day, Fig. 2I). 4. PDTC group was injected with PDTC (The pyrrolidine dithiocarbamate, an inhibitor of NF-κB, 150 mg/kg, Chen, et al., 2013) for 5 days (4 h hypoxia (10.8% O2) /day, Fig. 2E) or for 8 h hypoxia (8.2% O2, Fig. 2B,G). 5. An antagonist group (Fig. 2C,H) was treated with CP154,526 (an antagonist of CRHR1, 30 mg/kg, kindly donated by Pfizer Inc.USA). After exposure, rats were rapidly decapitated within half an hour at 14:00 -14:30 to minimize circadian rhythm effects.
China) weighing 180 ± 20 g were group-housed in the behavior lab 7 days for environmental adaptation before experiments. Rats in the hypoxia group were placed in a hypobaric chamber and exposed to hypobaric hypoxia of 7000 m altitude (~8.2% O 2 )for 1, 8, 24 h (Fig. 2A, B,C,F,G,H,J) or a prolonged hypoxia (PH), which mimicked at altitude of 5000 m (~10.8% O 2 ) for 2 or 5 days (4 h/per day, Fig. 2D,E,I) (Hao et al., 2015) . The normoxia group (Control, Con) was placed in the same chamber set at sea level (~ 21 % O 2 ). Rats were randomized into different groups. 1. The Control group was injected (ip) with 0.9% saline. 2. The Hypoxia group was injected with vehicle (0.9% saline) before hypoxia stress. 3. Dex group was injected with Dexamethasone (Dex, 500 µg/kg, ip) for 2 days (4 h hypoxia /day, Fig. 2I). 4. PDTC group was injected with PDTC (The pyrrolidine dithiocarbamate, an inhibitor of NF-κB, 150 mg/kg, Chen, et al., 2013) for 5 days (4 h hypoxia (10.8% O2) /day, Fig. 2E) or for 8 h hypoxia (8.2% O2, Fig. 2B,G). 5. An antagonist group (Fig. 2C,H) was treated with CP154,526 (an antagonist of CRHR1, 30 mg/kg, kindly donated by Pfizer Inc.USA). After exposure, rats were rapidly decapitated within half an hour at 14:00 -14:30 to minimize circadian rhythm effects.
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