November 19, 2020
Supplementary Materialsnutrients-11-02978-s001. (Hmox1) protein in the liver of weaned piglets with IUGR. In conclusion, IUGR decreased the antioxidant capacity of newborn and weaned piglets. Curcumin could efficiently improve the growth, increase hepatic antioxidant capacity, and upregulate Nrf2 and Hmox1 levels in the liver of IUGR weaned piglets. = 10/group, five males and five females), and IUGR piglets were randomly assigned to the IUGR and IC (curcumin supplementation) groups (= 10/group, five males and five females). The NBW and IUGR groups were fed with basal diets, and the NC and IC groups were fed with basal diets supplemented with 400 mg curcumin/kg until day 50. The diets supplemented with 400 mg curcumin/kg was according to the previous study . They exhibited that dietary supplementation of 400 mg curcumin/kg was more effective in improving the health status of weaned pigs. All piglets were housed individually at an ambient heat of 25C28 C and experienced free access to water. At 50 d of age, piglets were weighed after feed deprivation for 12 h to calculate total body-weight gain (BWG), and feed consumption was recorded daily by box to calculate total feed intake (FI) and feed conversion ratio (G:F, BWG:FI). The compositions of the diets are offered in Table S1. A total of 32 piglets with nearly similar body weight within group (eight piglets/group, half male and half female) were stunned by electric shock and killed by jugular bloodletting at the end of the test. 2.3. Test Collection At 0 d old in ARPC4 test 1 and 50 d old in test 2, bloodstream examples were obtained by jugular venipuncture and centrifuged in 3000 for 15 min in 4 C Noopept after that. The serum was kept at ?20 C to keep carefully the contents stable as well as for additional analyses. The piglets had been killed in the region of one piglet per group in order to avoid the effect of your time. In both tests, fresh liver cells samples (the same right lobe area) were immediately collected using ice cubes and then stored at ?80 C in order to avoid the degradation of RNA and proteins and for further analyses. 2.4. Analysis of Serum Guidelines Serum lipid peroxidation level was indicated by malondialdehyde concentration (MDA Concentration Screening Kit, no. A003), which is a byproduct of lipid peroxidation. Concentrations of MDA and hydrogen peroxide (H2O2 Concentration Testing Kit, no. A064-1), activities of total Noopept antioxidant capacity (TAOC Activity Testing Kit, no. A015-1), catalase (CAT Activity Testing Kit, no. A007-1), glutathione peroxidase (GSH-Px Activity Testing Kit, no. A005), and glutathione reductase (GR Activity Testing Kit, no. A062) in the serum were determined according to the manufacturers instructions of Nanjing Jiancheng Bioengineering Institute (Nanjing, Jiangsu Province, China). The detailed instructions of these screening kits are clearly described in our supplemental documents (https://doi.org/10.5281/zenodo.3520037). Serum activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were determined according to the earlier study (Selecta XL; Vital medical, Newton, MA, USA) . 2.5. Analysis of Liver Antioxidant Status The frozen liver samples (0.4 g) from ?80 C were homogenized having a handheld homogenizer (Pro 200; Pro Scientific Inc., Oxford, CT, USA) in 0.86% (w/v) ice-cold physiological saline (3.6 mL) or cells homogenate provided by the related diagnostic kit (Nanjing Jiancheng Bioengineering Institute, Jiangsu, China) according to the instructions of the manufacturer. The homogenate was centrifuged at 3500 for 15 min at 4 C and the supernatants were immediately collected and stored at ?20 C for measurement. Protein contents of liver were measured using the bicinchoninic acid (BCA) protein assay of Nanjing Jiancheng Bioengineering Institute (Nanjing, Jiangsu Province, China; BCA Assay Kits, no. A045-3). Protein oxidation in the liver was measured via the concentration of protein carbonyl (Personal computer Concentration Testing Kit, no. A087). Concentrations of MDA (MDA Concentration Testing Kit, no. A003), H2O2 (H2O2 Concentration Testing Kit, no. A064-1), glutathione (GSH Concentration Testing Noopept Kit, no. A006), oxidized glutathione (GSSG Concentration Testing Kit, no. A061-2), glutathione reductase (GR Concentration Testing Kit, no. A062), and activities of CAT (CAT Activity Testing Kit, no. A007-1), TAOC Noopept (TAOC Activity Testing Kit, no. A015-1), GSH-Px (GSH-Px Activity Testing Kit, no. A005), total nitric oxide synthase (TNOS Activity Testing Kit,.