Hepatic and Skeletal Muscle Autophagy Marker Levels in Rat Models of Prenatal and Postnatal Protein Restriction
<p>Body weight of pups on day 0 to week 3. Body weight of Con, Pre, and Pre + Post pups was recorded at birth and weeks 1, 2, and 3. Body weight of Pre and Pre + Post pups was significantly reduced compared with that of the Con group. Con, control; Pre, prenatal protein restriction; Pre + Post, prenatal and postnatal protein restriction. **, <span class="html-italic">p</span> < 0.0001; *, <span class="html-italic">p</span> < 0.05.</p> "> Figure 2
<p>Blood glucose level on week 3. The blood glucose level was measured before organ sampling. Blood glucose levels between the Con and Pre groups were not significantly different. Con, control; ns, not significant; Pre, prenatal protein restriction; Pre + Post, prenatal and postnatal protein restriction. ***, <span class="html-italic">p</span> < 0.005; **, <span class="html-italic">p</span> < 0.05.</p> "> Figure 3
<p>Western blot of the autophagic marker in livers of newborn pups (day 0). Protein expression of hepatic autophagy markers in newborn pups. Atg12 and LC3-II protein expression was significantly reduced in the Pre group compared to the Con group (<span class="html-italic">p</span> = 0.0015 and <span class="html-italic">p</span> = 0.01, respectively; <span class="html-italic">n</span> = 5 per group). Protein expression of the termination phase p62 significantly increased in the Pre group (<span class="html-italic">p</span> = 0.024). Bar columns represent the mean and error bars represent the standard deviation. Atg12, autophagy-related 12; Con, control; LC3-II, microtubule-associated protein light chain 3-II; ns, not significant; Pre, prenatal protein restriction. **, <span class="html-italic">p</span> < 0.005; *, <span class="html-italic">p</span> < 0.05.</p> "> Figure 4
<p>Western blot of the autophagic markers in livers of 3-week-old pups. Hepatic protein levels of LC3-I and LC3-II decreased in the Pre and Pre + Post groups compared with those in the Con group (<span class="html-italic">p</span> < 0.0001), whereas the level of beclin-1 increased in the 3-week-old Pre + Post pups compared with that in the Pre group pups (<span class="html-italic">p</span> = 0.0064), but it did not significantly differ from the levels in the Con group. Bar columns and error bars represent mean and standard deviation, respectively. Atg12, autophagy-related 12; Con, control; LC3-II, microtubule-associated protein light chain 3-II; ns, not significant; Pre, prenatal protein restriction; Pre + Post, prenatal and postnatal protein restriction. **, <span class="html-italic">p</span> < 0.005; ****, <span class="html-italic">p</span> < 0.0001.</p> "> Figure 5
<p>Western blot for skeletal muscle autophagic marker at week 3. Expression of autophagy regulator proteins beclin-1, LC3-II, and p62 decreased in the skeletal muscles of the Pre pups compared with that in the skeletal muscles of the Con pups (<span class="html-italic">p</span> = 0.04, <span class="html-italic">p</span> = 0.04, and <span class="html-italic">p</span> = 0.01, respectively). The levels of autophagy regulator proteins (Atg12, LC3-II, and p62) were also significantly lower in the Pre pups than in the Pre + Post pups (<span class="html-italic">p</span> = 0.02, <span class="html-italic">p</span> = 0.03, and <span class="html-italic">p</span> = 0.01, respectively). Atg12, autophagy-related 12; Con, control; LC3-II, microtubule-associated protein light chain 3-II; ns, not significant; Pre, prenatal protein restriction; Pre + Post, prenatal and postnatal protein restriction. *, <span class="html-italic">p</span> < 0.05.</p> "> Figure 6
<p>Ratio of LC3-II:LC3-I in the (<b>a</b>) liver and (<b>b</b>) skeletal muscles. In the liver, the ratio of LC3-II:LC3-I as autophagy markers was lower in Pre pups on day 0 and higher in Pre + Post pups than in the Pre and Con groups by week 3. Bar columns represent the mean, and error bars represent the standard deviation. Con, control; LC3-II, microtubule-associated protein light chain 3-II; ns, not significant; Pre, prenatal protein restriction; Pre + Post, prenatal and postnatal protein restriction. ***, <span class="html-italic">p</span> < 0.005; **, <span class="html-italic">p</span> < 0.05.</p> "> Figure 7
<p>Western blot of the ER stress markers in the livers of (<b>a</b>) newborn pups and (<b>b</b>) 3-week-old pups. Hepatic CHOP and BiP protein levels in the Con and Pre groups were not significantly different in newborn pup livers. Bar columns and error bars represent mean and standard deviation, respectively. Con, control; FGR, fetal growth restriction; ns, not significant; Pre + Post, prenatal and postnatal protein restriction.</p> "> Figure 8
<p>Western blot of skeletal muscle ER stress markers at week 3. Expression of ER stress marker proteins CHOP and BiP was not significantly different among the groups. Con, control; ns, not significant; Pre, prenatal protein restriction; Pre + Post, prenatal and postnatal protein restriction.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Animals
2.2. Prenatal and Postnatal Nutrition Management
2.3. Western Blotting
2.4. Statistical Analysis
3. Results
3.1. Body Weight and Glucose Level of the Rat Model
3.2. Hepatic and Skeletal Muscle Autophagy-Related Proteins
3.3. Stress Markers in the Liver and Skeletal Muscle
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
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Santosa, I.; Shoji, H.; Arai, Y.; Awata, K.; Tokita, K.; Shimizu, T. Hepatic and Skeletal Muscle Autophagy Marker Levels in Rat Models of Prenatal and Postnatal Protein Restriction. Nutrients 2023, 15, 3058. https://doi.org/10.3390/nu15133058
Santosa I, Shoji H, Arai Y, Awata K, Tokita K, Shimizu T. Hepatic and Skeletal Muscle Autophagy Marker Levels in Rat Models of Prenatal and Postnatal Protein Restriction. Nutrients. 2023; 15(13):3058. https://doi.org/10.3390/nu15133058
Chicago/Turabian StyleSantosa, Irena, Hiromichi Shoji, Yoshiteru Arai, Kentaro Awata, Kazuhide Tokita, and Toshiaki Shimizu. 2023. "Hepatic and Skeletal Muscle Autophagy Marker Levels in Rat Models of Prenatal and Postnatal Protein Restriction" Nutrients 15, no. 13: 3058. https://doi.org/10.3390/nu15133058