Cell experiment:

The PPARβ activator GW0742 and the RXR activator 9-cis-retinoic acid are dissolved in DMSO. The final DMSO concentration des not exceed 0.5% v/v, and this concentration is used in control wells. For each culture plate, one row of wells is treated with 500 μM glutamate. These wells serve as a positive control and for normalisation of data. Cell death (toxicity) is assessed by using an assay designed to measure lactate dehydrogenase (LDH) release[2].

Animal experiment:

Male CD mice (25-35 g) are housed in a controlled environment and provided with standard rodent chow and water. Mice are randomized into four experimental groups: bleomycin-treated group: mice are subjected to lung injury induced by intratracheal instillation of bleomycin and treated daily via intraperitoneal injection with vehicle of GW0742 (10% dimethylsulfoxide (OMSO, 1 mL/kg), 1 h after BLEO instillation (n = 15). GW0742 group: identical to bleomycin-treated group but mice are treated daily with GW0742 (0.3 mg/kg, 1h after BLEO instillation) via intraperitoneal injection (n = 15). Sham-operated mice + vehicle group: animals are subjected to the identical surgical procedure but receive intratracheal instillation of saline (0.9%) instead of BLEO and are treated daily with the vehicle of GW0742 (10% dimethylsulfoxide (DMSO), 1 mL/kg, i.p.), 1 h after saline instillation (n = 15). Sham-operated mice + GW0742 group: identical to sham + vehicle group but mice are treated daily with GW0742 (0.3 mg/kg, 1 h after saline instillation) via intraperitoneal injection (n = 15)[3].

GW0742 is a synthetic, potent and selective PPAR-β/δ agonist. PPARs are ligand-dependent transcription factors which are involved in many physiological processes, such as inflammation and energy homeostasis. PPAR-β/δ is one of three PPARs in the nuclear hormone receptor superfamily that are collectively involved in the control of lipid homoeostasis among other functions. GW0742 can attenuate the increase of PARP activity that caused by SAO shock. GW0742 is also able to prevent radiation-induced brain injury in C57Bl/6 wild-type (WT) and PPARδ knockout (KO) mice. Dietary GW0742 can prevent the acute increase in IL-1β mRNA and ERK phosphorylation measured at 3 h after a single 10-Gy dose of WBI as well as the increase in the number of activated hippocampal microglia 1 week after WBI.

Reference

[1].Rosanna Di Paola, Emanuela Esposito, Emanuela Mazzon, Irene Paterniti, Maria Galuppo, Salvatore Cuzzocrea. GW0742, a selective PPAR-β/δ agonist, contributes to the resolution of inflammation after gut ischemia/reperfusion injury. August 2010. Journal of Leukocyte Biology. vol. 88 no. 2 291-301
[2].Caroline I. Schnegg, Dana Greene-Schloesser, Mitra Kooshki, Valerie S. Payne, Fang-Chi Hsud, Mike E. Robbins. hippocampal-dependent cognitive impairment after whole-brain irradiation. Free Radical Biology and Medicine. Volume 61, August 2013, Pages 1–9.