[en] [en] INTRODUCTION: Maternal dietary protein intake plays a vital role in pregnancy outcomes. However, the impact of popular high-protein diets, often accompanied by carbohydrate restriction, remains poorly understood. We investigated the effect of different levels of maternal protein during pregnancy on embryo implantation, placental development, and fetal growth in mice.
METHODS: Pregnant C57BL/6 mice were fed isocaloric purified diets containing 10% (low protein, LP), 20% (medium protein, MP), or 40% (high protein, HP) protein. Implantation was assessed on day 6.5, and gestational outcomes were analyzed on day 18.5.
RESULTS: Maternal protein did not affect implantation number or litter size. However, HP intake reduced fetal weight and placental efficiency. Morphologically, HP intake decreased the placental labyrinth zone/total area ratio compared to LP. It was further revealed that maternal HP intake reduced fetal serum glucose, increased fetal serum urea nitrogen, and activated hepatic gluconeogenic and amino acid catabolic pathways compared to MP. Mechanistically, HP intake induced placental oxidative stress, evidenced by enhanced malondialdehyde content, and decreased the activity of placental catalase, superoxide dismutase, and glutathione peroxidase. The expression of genes involved in antioxidant defense and amino acid transport was also suppressed in placenta. Moreover, maternal HP intake reduced ATP production, lowered mitochondrial DNA copy number, and downregulated gene expression of electron transport chain components.
DISCUSSION: Excessive maternal protein intake under isoenergetic conditions disrupts placental redox homeostasis and mitochondrial function, and impairs nutrient transfer. These alterations compromise placental efficiency and fetal growth, highlighting the potential risks associated with high-protein dietary patterns during pregnancy.
Disciplines :
Zoology
Author, co-author :
Zhao, Ying ; Université de Liège - ULiège > TERRA Research Centre
Cai, Long; Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing, 100097, China
He, Liyuan; Key Laboratory of Feed Biotechnology of Ministry of Agriculture and Rural Affairs, Institute of Feed Research, Chinese Academy of Agricultural Sciences, Beijing, 100081, China, College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang, 453003, China
Gao, Ge; Key Laboratory of Feed Biotechnology of Ministry of Agriculture and Rural Affairs, Institute of Feed Research, Chinese Academy of Agricultural Sciences, Beijing, 100081, China
Schroyen, Martine ; Université de Liège - ULiège > TERRA Research Centre > Animal Sciences (AS)
Pi, Yu; Key Laboratory of Feed Biotechnology of Ministry of Agriculture and Rural Affairs, Institute of Feed Research, Chinese Academy of Agricultural Sciences, Beijing, 100081, China
Sun, Wenjuan; Key Laboratory of Feed Biotechnology of Ministry of Agriculture and Rural Affairs, Institute of Feed Research, Chinese Academy of Agricultural Sciences, Beijing, 100081, China
Ming, Dongxu; Key Laboratory of Feed Biotechnology of Ministry of Agriculture and Rural Affairs, Institute of Feed Research, Chinese Academy of Agricultural Sciences, Beijing, 100081, China
Li, Yanpin; Key Laboratory of Feed Biotechnology of Ministry of Agriculture and Rural Affairs, Institute of Feed Research, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. Electronic address: liyanpin@caas.cn
Li, Xilong; Key Laboratory of Feed Biotechnology of Ministry of Agriculture and Rural Affairs, Institute of Feed Research, Chinese Academy of Agricultural Sciences, Beijing, 100081, China. Electronic address: lixilong@caas.cn
Language :
English
Title :
Maternal high-protein diet impairs fetal growth through placental dysfunction in mice.
CSC - China Scholarship Council NSCF - National Natural Science Foundation of China
Funding text :
This research was supported by the National Natural Science Foundation of China (Grant number 32272908 ). The author Ying Zhao is grateful for the financial support provided by the China Scholarship Council program (Grant number 202403250107 ).
R. Elango, R.O. Ball, Protein and amino acid requirements during pregnancy, Adv. Nutr. 7(4) (2016) 839S-844S. https://doi.org/10.3945/an.115.011817.
S.C. Langley-Evans, S.J. Welham, R.C. Sherman, A.A. Jackson, Weanling rats exposed to maternal low-protein diets during discrete periods of gestation exhibit differing severity of hypertension, Clin. Sci. 91(5) (1996) 607-615. https://doi.org/10.1042/cs0910607.
D.S. Fernandez-Twinn, S. Ekizoglou, A. Wayman, C.J. Petry, S.E. Ozanne, Maternal low-protein diet programs cardiac β-adrenergic response and signaling in 3-mo-old male offspring, Am. J. Physiol. Regul. Integr. Comp. Physiol. 291(2) (2006) R429-R436. https://doi.org/10.1152/ajpregu.00608.2005.
C.J. Petry, M.W. Dorling, D.B. Pawlak, S.E. Ozanne, C.N. Hales, Diabetes in old male offspring of rat dams fed a reduced protein diet, Int. J. Exp. Diabetes. Res. 2(2) (2001) 139-143. https://doi.org/10.1155/edr.2001.139.
V. Zohdi, K. Lim, J.T. Pearson, M.J. Black, Developmental programming of cardiovascular disease following intrauterine growth restriction: findings utilising a rat model of maternal protein restriction, Nutrients. 7(1) (2014) 119-152. https://doi.org/10.3390/nu7010119.
C. Guzman, R. Cabrera, M. Cardenas, F. Larrea, P.W. Nathanielsz, E. Zambrano, Protein restriction during fetal and neonatal development in the rat alters reproductive function and accelerates reproductive ageing in female progeny, J. Physiol. 572(1) (2006) 97-108. https://doi.org/10.1113/jphysiol.2005.103903.
J. Yang, Q. Chang, X. Tian, B. Zhang, L. Zeng, H. Yan, S. Dang, Y.-H. Li, Dietary protein intake during pregnancy and birth weight among Chinese pregnant women with low intake of protein, Nutr. Metab. (Lond.) 19(1) (2022) 43. https://doi.org/10.1186/s12986-022-00678-0.
H. Zhang, A.M. Senior, C. Saner, N.A. Koemel, S.J. Simpson, D. Raubenheimer, B.L. Heitmann, Maternal protein intake during pregnancy and obesity risk in mothers and offspring: a prospective cohort study, Am. J. Clin. Nutr. (2025). https://doi.org/10.1016/j.ajcnut.2025.04.011.
D. Mikolainis, Impact of the Higgins Nutrition Intervention Program on birth weight: a within-mother analysis, J. Am. Diet. Assoc. 89(8) (1989) 1097-1103.
K. Andreasyan, A.-L. Ponsonby, T. Dwyer, R. Morley, M. Riley, K. Dear, J. Cochrane, Higher maternal dietary protein intake in late pregnancy is associated with a lower infant ponderal index at birth, Eur. J. Clin. Nutr. 61(4) (2007) 498-508. https://doi.org/10.1038/sj.ejcn.1602552.
C. Rehfeldt, L. Lefaucheur, J. Block, B. Stabenow, R. Pfuhl, W. Otten, C.C. Metges, C. Kalbe, Limited and excess protein intake of pregnant gilts differently affects body composition and cellularity of skeletal muscle and subcutaneous adipose tissue of newborn and weanling piglets, Eur. J. Nutr. 51(2) (2012) 151-165. https://doi.org/10.1007/s00394-011-0201-8.
J.C. Cross, L. Mickelson, Nutritional influences on implantation and placental development, Nutr. Rev. 64(suppl_2) (2006) S12-S18. https://doi.org/10.1301/nr.may.s12-s18.
I. Cetin, G. Alvino, Intrauterine growth restriction: implications for placental metabolism and transport. A review, Placenta. 30 (2009) 77-82. https://doi.org/10.1016/j.placenta.2008.12.006.
A.L. Fowden, A. Sferruzzi-Perri, P. Coan, M. Constancia, G. Burton, Placental efficiency and adaptation: endocrine regulation, J. Physiol. 587(14) (2009) 3459-3472. https://doi.org/10.1113/jphysiol.2009.173013.
P.D. Gluckman, M.A. Hanson, C. Cooper, K.L. Thornburg, Effect of in utero and early-life conditions on adult health and disease, New Engl. J. Med. 359(1) (2008) 61-73. https://doi.org/10.1056/NEJMra0708473.
O. Holland, M.D. Nitert, L.A. Gallo, M. Vejzovic, J.J. Fisher, A.V. Perkins, Placental mitochondrial function and structure in gestational disorders, Placenta. 54 (2017) 2-9. https://doi.org/10.1016/j.placenta.2016.12.012.
Z.C. Luo, W.D. Fraser, P. Julien, C.L. Deal, F. Audibert, G.N. Smith, X. Xiong, M. Walker, Tracing the origins of "fetal origins" of adult diseases: programming by oxidative stress?, Med. Hypotheses. 66(1) (2006) 38-44. https://doi.org/10.1016/j.mehy.2005.08.020.
S. Gupta, A. Agarwal, R.K. Sharma, The role of placental oxidative stress and lipid peroxidation in preeclampsia, Obstet. Gynecol. Surv. 60(12) (2005) 807-816. https://doi.org/10.1097/01.ogx.0000193879.79268.59.
T.-H. Hung, J.N. Skepper, D.S. Charnock-Jones, G.J. Burton, Hypoxia-reoxygenation: a potent inducer of apoptotic changes in the human placenta and possible etiological factor in preeclampsia, Circul. Res. 90(12) (2002) 1274-1281. https://doi.org/10.1161/01.res.0000024411.22110.aa.
J.O. Lo, M.C. Schabel, J. Gaffney, K.S. Lewandowski, C.D. Kroenke, C.T. Roberts Jr, B.P. Scottoline, A.E. Frias, E.L. Sullivan, V.H. Roberts, Impaired placental hemodynamics and function in a non-human primate model of gestational protein restriction, Sci. Rep. 13(1) (2023) 841. https://doi.org/10.1038/s41598-023-28051-y.
A. Trichopoulou, T. Psaltopoulou, P. Orfanos, C. Hsieh, D. Trichopoulos, Low-carbohydrate-high-protein diet and long-term survival in a general population cohort, Eur. J. Clin. Nutr. 61(5) (2007) 575-581. https://doi.org/10.1038/sj.ejcn.1602557.
P. Clifton, D. Condo, J. Keogh, Long term weight maintenance after advice to consume low carbohydrate, higher protein diets-a systematic review and meta analysis, Nutr. Metab. Cardiovasc. Dis. 24(3) (2014) 224-235. https://doi.org/10.1016/j.numecd.2013.11.006.
E. Mellott, D. Moronge, K. Backer, G. Cooper, S. Ogbi, J. Maher, J. Faulkner, Mid-late Gestation Eplerenone Treatment Improves Maternal Cardiovascular, But Not Fetal, Outcomes in a Mouse Model of Leptin-induced Preeclampsia, Physiology. 40(S1) (2025) 0252. https://doi.org/10.1152/physiol.2025.40.S1.0252.
C.E. Hayward, S. Lean, C.P. Sibley, R.L. Jones, M. Wareing, S.L. Greenwood, M.R. Dilworth, Placental adaptation: what can we learn from birthweight: placental weight ratio?, Front. Physiol. 7 (2016) 28. https://doi.org/10.3389/fphys.2016.00028.
Z. Dai, Z. Wu, S. Jia, G. Wu, Analysis of amino acid composition in proteins of animal tissues and foods as pre-column o-phthaldialdehyde derivatives by HPLC with fluorescence detection, J. Chromatogr. B. Analyt. Technol. Biomed. Life. Sci. 964 (2014) 116-127. https://doi.org/10.1016/j.jchromb.2014.03.025.
J. Wu, M. Zhang, H. Zhang, M. Sheng, J. Sun, F. Wu, H. Gao, L. Chen, Z. Li, Q. Tian, PMN-MDSC: A Culprit Behind Immunosenescence and Increased Susceptibility to Clostridioides difficile Infection During Aging, Engineering. 42 (2024) 59-73. https://doi.org/10.1016/j.eng.2024.06.014.
M.-y. He, G. Wang, S.-s. Han, Y. Jin, H. Li, X. Wu, Z.-l. Ma, X. Cheng, X. Tang, X. Yang, Nrf2 signalling and autophagy are involved in diabetes mellitus-induced defects in the development of mouse placenta, Open. Biol. 6(7) (2016) 160064. https://doi.org/10.1098/rsob.160064.
J.S. Eaton, Z.P. Lin, A.C. Sartorelli, N.D. Bonawitz, G.S. Shadel, Ataxia-telangiectasia mutated kinase regulates ribonucleotide reductase and mitochondrial homeostasis, J. Clin. Invest. 117(9) (2007) 2723-2734. https://doi.org/10.1172/JCI31604.
M. Zhao, S. Liu, C. Wang, Y. Wang, M. Wan, F. Liu, M. Gong, Y. Yuan, Y. Chen, J. Cheng, Mesenchymal stem cell-derived extracellular vesicles attenuate mitochondrial damage and inflammation by stabilizing mitochondrial DNA, ACS. Nano. 15(1) (2020) 1519-1538. https://doi.org/10.1021/acsnano.1c09895.
E.D. Watson, J.C. Cross, Development of structures and transport functions in the mouse placenta, Physiology. 20(3) (2005) 180-193. https://doi.org/10.1152/physiol.00001.2005.
P.K. Ajuogu, M. Wolden, J.R. McFarlane, R.A. Hart, D.J. Carlson, T. Van der Touw, N.A. Smart, Effect of low-and high-protein maternal diets during gestation on reproductive outcomes in the rat: a systematic review and meta-analysis, J. Anim. Sci. 98(1) (2020) skz380. https://doi.org/10.1093/jas/skz380.
K.E. Brett, Z.M. Ferraro, J. Yockell-Lelievre, A. Gruslin, K.B. Adamo, Maternal-fetal nutrient transport in pregnancy pathologies: the role of the placenta, Int. J. Mol. Sci. 15(9) (2014) 16153-16185. https://doi.org/10.3390/ijms150916153.
K.K.S. Bhasin, A. van Nas, L.J. Martin, R.C. Davis, S.U. Devaskar, A.J. Lusis, Maternal low-protein diet or hypercholesterolemia reduces circulating essential amino acids and leads to intrauterine growth restriction, Diabetes. 58(3) (2009) 559-566. https://doi.org/10.2337/db07-1530.
J.K. Cleal, E.M. Lofthouse, B.G. Sengers, R.M. Lewis, A systems perspective on placental amino acid transport, J Physiol. 596(23) (2018) 5511-5522. https://doi.org/10.1113/JP274883. Epub 2018 Sep 7.
M.A. Underwood, W.M. Gilbert, M.P. Sherman, Amniotic fluid: not just fetal urine anymore, J. Perinatol. 25(5) (2005) 341-348. https://doi.org/10.1038/sj.jp.7211290.
W. Schmidt, The amniotic fluid compartment: the fetal habitat, Adv. Anat. Embryol. Cell. Biol. 127 (1992) 1-100. https://doi.org/10.1007/978-3-642-77300-6.
J. Cross, How to make a placenta: mechanisms of trophoblast cell differentiation in mice-a review, Placenta. 26 (2005) S3-S9. https://doi.org/10.1016/j.placenta.2005.01.015.
P. Coan, N. Conroy, G. Burton, A. Ferguson-Smith, Origin and characteristics of glycogen cells in the developing murine placenta, Dev. Dyn. 235(12) (2006) 3280-3294. https://doi.org/10.1002/dvdy.20981.
E.H. Joo, Y.R. Kim, N. Kim, J.E. Jung, S.H. Han, H.Y. Cho, Effect of endogenic and exogenic oxidative stress triggers on adverse pregnancy outcomes: preeclampsia, fetal growth restriction, gestational diabetes mellitus and preterm birth, Int. J. Mol. Sci. 22(18) (2021) 10122. https://doi.org/10.3390/ijms221810122.
L. Myatt, X. Cui, Oxidative stress in the placenta, Histochem. Cell. Biol. 122 (2004) 369-382. https://doi.org/10.1007/s00418-004-0677-x.
O. Ighodaro, O. Akinloye, First line defence antioxidants-superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX): Their fundamental role in the entire antioxidant defence grid, Alex. J. Med. 54(4) (2018) 287-293. https://doi.org/10.1016/j.ajme.2017.09.001.
T. Nguyen, P. Nioi, C.B. Pickett, The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress, J. Biol. Chem. 284(20) (2009) 13291-13295. https://doi.org/10.1074/jbc.R900010200.
D. Poidatz, E. Dos Santos, F. Duval, H. Moindjie, V. Serazin, F. Vialard, P. De Mazancourt, M.-N. Dieudonne, Involvement of estrogen-related receptor-γ and mitochondrial content in intrauterine growth restriction and preeclampsia, Fertil. Steril. 104(2) (2015) 483-490. https://doi.org/10.1016/j.fertnstert.2015.05.005.
A. Musatov, N.C. Robinson, Susceptibility of mitochondrial electron-transport complexes to oxidative damage. Focus on cytochrome c oxidase, Free Radic. Res. 46(11) (2012) 1313-1326. https://doi.org/10.3109/10715762.2012.717273.
M. Adebayo, S. Singh, A.P. Singh, S. Dasgupta, Mitochondrial fusion and fission: the fine-tune balance for cellular homeostasis, FASEB. J. 35(6) (2021) e21620. https://doi.org/10.1096/fj.202100067R.
