Excessive backfat of sows at mating promotes oxidative stress and up-regulates mitochondrial-mediated apoptotic pathway in the full-term placenta

Impaired Mitochondrial Function Results from Oxidative Stress in the Full-Term Placenta of Sows with Excessive Back-Fat

“…Consistently, we found that elevated lipid promoted the activation of NF-κB associated with increased ROS and impaired mitochondrial function and structure in cytotrophoblasts. Moreover, our previous studies showed that excessive back-fat is associated with increased activation of Mark4 and reduced activity and expression of AMPK in pig term placenta, indicating a potential mechanism for increased placental inflammation and oxidative stress [8,17]. In this trial, increased activation of microtubule affinity-regulating kinase 4 (Mark4) and reduced activation of AMPK were also observed in cultured trophoblast cells treated with high fatty acid, and addition of vitamin E alleviated these effects, suggesting that Mark4 and AMPK may be involved in regulation of ROS-mediated mitochondrial abnormalities in pig placenta induced by increased back-fat.…”

“…All the sows were multiparous with same parity (Parity = 2) and inseminated with pooled semen doses containing 3-4 billion sperm cells to produce purebred litters. After the weaning period with a corn soybean diet (3200 Kal/kg of metabolizable energy, 18% of crude protein and 0.9% lysine), the females were dichotomized based on BF (back-fat thickness) at mating into BFI (15-20 mm, n = 10) and BFII (21-27 mm, n = 10) as described previously [17]. All the sows were housed together in passively ventilated collective pens with concrete slatted floors at the Research Farm of Nan Jing Agricultural University.…”

“…The concentrations of triglyceride (TG) and nonesterified fatty acids (NEFA) in plasma samples were determined by using Beckman AU5811 analyzer (Beckman-Coulter, Inc.250S.Kraemer Boulevard Brea, CA, USA) as previously reported [8]. A commercially available porcine specific ELISA kit (RD SYSTEMS, Minneapolis, MN, USA) and an Amplex Red hydrogen peroxide assay kit (Invitrogen, Carlsbad, CA, USA) were utilized to measure plasma leptin and hydrogen peroxide (H 2 O 2 ) content, respectively, according to the manufacturer's instructions as previously described [5,17]. All blood samples were determined in duplicate in single assay.…”

“…Fluorescent probe JC-1 (KeyGen BioTECH, Nanjing, China) was used to estimate mitochondrial membrane potential (∆Ψm) as previously described [17,23]. Cytotrophoblasts were harvested, washed with cold PBS, and incubated at 37 • C for 15 min with 5 µg/mL JC-1.…”

“…For mitochondrial complexes activity and ATP production rate measurement, mitochondria was isolated from placental villous tissue or cell homogenate using Mitochondria Isolation Kit (KeyGen BioTECH, Nanjing, China) as previously described [17]. Briefly, placental samples (200 mg) or cytotrophoblasts were homogenized in ice-cold isolation buffer.…”