General Obstetrics and Gynecology: Obstetrics
Novel peptides prevent alcohol-induced spatial learning deficits and proinflammatory cytokine release in a mouse model of fetal alcohol syndrome

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Objective

Previously, the novel peptides NAPVSIPQ and SALLRSIPA were shown to prevent alcohol-induced fetal death and growth abnormalities in a mouse model of fetal alcohol syndrome. This study evaluated whether these peptides could prevent long-term alcohol-induced learning abnormalities. In addition, because specific cytokines are known to effect long-term potentiation, a model of learning at the molecular level, we studied the effect of these novel peptides on tumor necrosis factor–α, interleukin-6, and interferon-γ levels.

Study design

We used a well-characterized mouse model of fetal alcohol syndrome. Pregnant mice were injected on day 8 with alcohol (0.03 mL/kg) or placebo. Pretreatment with NAPVSIPQ + SALLRSIPA (20 μg) or placebo was given 30 minutes before alcohol. Embryos were removed after 6 hours, at which time cytokine, tumor necrosis factor–α, interleukin-6, and interferon-γ levels were measured with enzyme-linked immunoassays. To test spatial learning, adult offspring from litters that were treated with alcohol, control, NAPVSIPQ + SALLRSIPA then alcohol, or NAPVSIPQ + SALLRSIPA alone were evaluated for latency to find a hidden platform in the Morris water maze.

Results

Alcohol treatment increased tumor necrosis factor–α levels versus control levels (50.0 ± 3.5 pg/mL vs 32.7 ± 2.4 pg/mL; P < .001). NAPVSIPQ + SALLRSIPA pretreatment prevented this increase (39.9 9 ± 2.8 pg/mL; P ≤ .01), with levels similar to control (P = .1). Similarly, alcohol increased interleukin-6 levels versus control levels (22.6 ± 1.4 pg/mL vs 17.3 ± 0.6 pg/mL; P < .001), and NAPVSIPQ + SALLRSIPA prevented this increase (19.1 ± 1.0 pg/mL; P ≤ .02), with levels similar to control levels (P = .2). Interferon-γ levels were not different among the 3 groups (alcohol, 14.6 ± 4.9 pg/mL; control, 17.9 ± 6.6 pg/mL; alcohol + NAPVSIPQ + SALLRSIPA, 13.6 ± 4.9 pg/mL; P = .2). In the Morris water maze, alcohol-treated groups did not learn over the 7-day trial compared with the control group (P = .001). Groups that were pretreated with NAPVSIPQ + SALLRSIPA then alcohol learned significantly, which was similar to the control group. Groups that were treated with only NAPVSIPQ + SALLRSIPA learned significantly earlier, with the shortest latency once learning commenced.

Conclusion

The peptides, NAPVSIPQ + SALLRSIPA, prevented the alcohol-induced spatial learning deficits and attenuated alcohol-induced proinflammatory cytokine increase in a model of fetal alcohol syndrome. This study demonstrates the peptides' significant in vivo efficacy with long-lasting effects obtained after prenatal administration.

Section snippets

Material and methods

We used a well-characterized mouse model of FAS.13 C57Bl6/J female mice (Jackson Laboratories, Bar Harbor, Maine) were kept under a 12-hour light, 12-hour dark regimen, with food and water available at all times. The mice received humane animal care in compliance with the “Guideline for Care and Use of Experimental Animals.” Six-week-old females (21-24 g) were mated with C57Bl6/J males for 4 hours. On gestational day 8, we injected pregnant mice intraperitoneally with 25% ethyl alcohol in

Results

In the learning paradigm, offspring from the alcohol-treated litters demonstrated no evidence of learning over the 7-day trial (Figure). In contrast, animals from the control litters decreased their latency 50% by the fifth day (P<.001). Males from the litters who were pretreated with NAP + ADNF-9 and then given alcohol also significantly learned, with a learning curve not different from that of the control at all time points that were tested. The offspring from litters that were treated with

Comment

This study showed that alcohol treatment increases proinflammatory cytokine levels and produces spatial learning deficits in affected offspring. The data also imply that the peptides, NAP + ADNF-9, prevent alcohol-induced increases in proinflammatory cytokines and protect against alcohol-induced spatial learning deficits. In addition, this study suggests that treatment with NAP + ADNF-9 alone enhanced learning in normal mice, when compared with control. The data are consistent with previous work in

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