The synthetic triterpenoid 2-cyano-3,12-dioxooleana-1,9(11 (link))-dien-28-oic acid (CDDO) along with chemically modified derivatives, CDDO-Me (methyl ester, also known as bardoxolone methyl, BARD) and CDDO-EA (ethyl amide) are non-cytotoxic, highly multifunctional and orally available drugs that have been studied as anti-inflammatory and anti-oxidation agents in vivo and in vitro (43 (link)–45 (link)). Because ethyl amide derivative of a synthetic triterpenoid, CDDO-EA, has been reported to have enhanced pharmacodynamic activity in mouse assays compared to CDDO-Me (46 (link)), CDDO-EA (400 mg/kg diet) (provided from Reata Pharmaceuticals, Irving Texas, and Dr. Michael Sporn, Dartmouth Medical School, NH, USA) was first dissolved in an oily vehicle and prepared into chow pellets (Lab Diet #5002) by Purina (Purina-Mills, Richmond, IN, USA) (47 (link)). Group of mice were continuously fed a CDDO-EA diet or Control diet (Lab Diet #5002) for 3 days prior to radiation exposure.
Lab diet 5002
Manufactured by Purina
Sourced in United States
Lab Diet #5002 is a specialized laboratory animal feed formulated to meet the nutritional requirements of laboratory rodents. It is designed to provide a consistent and balanced diet for use in research and experimental settings.
Automatically generated - may contain errors
5 protocols using lab diet 5002
1
Triterpenoid CDDO-EA Anti-inflammatory Protocol
2
Inhalation Exposure of SPF Rats to 3-MBCF
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Male and female specific-pathogen-free (SPF) SD rats aged 6–7 weeks were purchased (Orient Bio Inc., South Korea) and acclimatized for 11 days in polycarbonate cage with SPF conditions before the grouping. Exposures were conducted in inhalation chambers (Model No. SIS-20RG, Sibata Co., Japan) with individual wire mesh cages, three for each concentration of 3-MBCF plus a control chamber with HEPA filtered clean air under SPF laboratory conditions. The ambient temperature and relative humidity of the chamber was 22 ± 3 °C and 50 ± 20%, respectively, with a 12:12 h light:dark cycle (lights on at 8:00 am) with 150 ∼ 300 lx of illumination. The rats received rodent chow (LabDiet 5002, Purina Mills., St. Louis, MO, USA) and tap water ad libitum. All animal protocols described in this study were approved by the Committee on Animal Research Committee of the Occupational Safety and Health Research Institute.
3
ACE2 Transgenic Mouse Models for COVID-19 Research
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Male and female B6.Cg-Tg(K18-ACE2)2Prlmn/J (#034860, Jackson Laboratory), C57BL/6 (B6; Jackson Laboratory), Ch25h−/− (#016263, Jackson Laboratory) (5 ), and Gpr183−/− ((19 (link)), kindly provided by Vanja Lazarevic) mice, ~22–26g in weight, were used. The light/dark cycle was set at 12/12 hours, and mice were fed Purina Lab Diet #5002 and provided water ad libitum. Animal care and housing met AAALAC International guidelines, the Guide for Care and Use of Laboratory Animals (National Research Council), and requirements as stated by the U.S. Department of Agriculture through the Animal Welfare Act. All experiments were performed in compliance with an animal study proposal approved by the NIAID Animal Care and Use Committee or the NIEHS Animal Care and Use Committee.
4
Triterpenoid CDDO-EA Anti-inflammatory Protocol
5
Inducing Metabolic Acidosis in Rodents
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Metabolic acidosis was induced via supplementation of rodent diet with 2% ammonium chloride (NH4Cl‐acidosis; NH4Cl‐A) ad libitum for up to 7 days. Ammonium chloride supplemented Labdiet® 5002 was formulated and manufactured by Purina Mills and distributed by Scott Pharma Solutions. HCl‐acidosis (HCl‐A) was induced by supplementing Labdiet 5010 with 1 ml/g 0.4 N HCl as described (Lee et al., 2009 (link)). Blood was collected from mice by tapping the retro‐orbital sinus under light anesthesia with a heparinized capillary pipette from which blood pH and s[HCO3−] were measured utilizing iSTAT® G3+ Cartridges (Abbott Labs). The volume and pH of dark cycle urine, collected under mineral oil from groups of 4–5 mice housed overnight in a metabolic cage with free access to food (powdered) and water, was determined the following morning. Urine pH was first estimated with pH strips (Whatman™Type CS pH indicator strips, 10SG Urine Reagent Strips; Thermo Fischer Scientific), and then confirmed using a calomel pH electrode (Table 1 ).
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