Pomegranates (POM), rich with polyphenol anti-oxidants, has been demonstrated to attenuate oxidative stress and apoptosis in human placental trophoblasts. Maternal and fetal inflammation has a central role in fetal brain injury. Infection/inflammation activates cell death pathways (apoptosis) and inflammatory responses through induction of caspase 3, oxidative stress and NF-kB pathways. We sought to determine if maternal pomegranate juice supplementation attenuates the activation of apoptosis and inflammatory pathways in response to simulated inflammation.
Pregnant rats (n=24) were provided ad libitum drinking water (H20) from throughout pregnancy. Beginning at day 11 of gestation, one half of the dams were provided water supplemented with pomegranate juice (5cc per day). At day 18, both groups of dams were randomized to receive injections of i.p. LPS (100 ug/kg) or saline (SAL). Rats were sacrificed 4 hours following LPS/ saline injection. Fetal brains were collected from the 4 treatment groups (H20/SAL, H20/LPS, POM/SAL, POM/LPS). We used one fetal brain from each dam, resulting in 6 brains from 6 different dams in each group. Fetal brain and placenta caspase 3 active form (af), NF-kB p65, neuronal nitric oxide synthase (phospho-nNos) and protein levels of interleukin (IL)-6, CCL2 and TNFα were determined by western blot analysis
Maternal LPS at e18 significantly (p<0.05) increased fetal brain caspase 3 af, NFkB p65, and phospho-nNOS protein levels compared to the control group (caspase 3 af 0.26 ± 0.01 vs. 0.20 ± 0.01 u; NFkB p65 0.24 ± 0.01 vs. 0.1 ± 0.01 u; phospho-Nnos 0.23 ± 0.01 vs. 0.11 ± 0.01 u) as well as fetal brain pro-inflammatory cytokines (IL-6 0.25 ± 0.01 vs. 0.09 ± 0.01 u; TNFα 0.26 ± 0.01 vs. 0.12 ± 0.01 u; CCL2 0.23 ± 0.01 vs. 0.1 ± 0.01 u). Maternal POM juice (POM/LPS) significantly (p < 0.05) reduced fetal brain responses to maternal LPS: caspase 3 af (0.2 ± 0.01 u), NFkB p65 (0.22 ± 0.01 u) and phospho- nNos (0.19 ± 0.01 u) as well as brain pro-inflammatory cytokines (IL-6 0.21 ± 0.01 u; TNFα 0.22 ± 0.01 u; CCL2 0.2 ± 0.01 u).
Maternal POM juice supplementation may attenuate maternal inflammation-induced fetal brain apoptosis, oxidative stress and inflammation. We speculate that the POM polyphenols have potent anti-inflammatory properties.
© 2016 Published by Elsevier Inc.