, Volume 61, Issue 8, pp 1862–1876 | Cite as

Folate treatment of pregnant rat dams abolishes metabolic effects in female offspring induced by a paternal pre-conception unhealthy diet

  • Jian Li
  • Yong-Ping Lu
  • Oleg Tsuprykov
  • Ahmed A. Hasan
  • Christoph Reichetzeder
  • Mei Tian
  • Xiao Li Zhang
  • Qin Zhang
  • Guo-Ying Sun
  • Jingli Guo
  • Mohamed M. S. Gaballa
  • Xiao-Ning Peng
  • Ge Lin
  • Berthold HocherEmail author



Paternal high-fat diet prior to mating programmes impaired glucose tolerance in female offspring. We examined whether the metabolic consequences in offspring could be abolished by folate treatment of either the male rats before mating or the corresponding female rats during pregnancy.


Male F0 rats were fed either control diet or high-fat, high-sucrose and high-salt diet (HFSSD), with or without folate, before mating. Male rats were mated with control-diet-fed dams. After mating, the F0 dams were fed control diet with or without folate during pregnancy.


Male, but not female offspring of HFSSD-fed founders were heavier than those of control-diet-fed counterparts (p < 0.05 and p = 0.066 in males and females, respectively). Both male and female offspring of HFSSD-fed founders were longer compared with control (p < 0.01 for both sexes). Folate treatment of the pregnant dams abolished the effect of the paternal diet on the offspring’s body length (p ˂ 0.05). Female offspring of HFSSD-fed founders developed impaired glucose tolerance, which was restored by folate treatment of the dams during pregnancy. The beta cell density per pancreatic islet was decreased in offspring of HFSSD-fed rats (−20% in male and −15% in female F1 offspring, p ˂ 0.001 vs controls). Folate treatment significantly increased the beta cell density (4.3% and 3.3% after folate supplementation given to dams and founders, respectively, p ˂ 0.05 vs the offspring of HFSSD-fed male rats). Changes in liver connective tissue of female offspring of HFSSD-fed founders were ameliorated by treatment of dams with folate (p ˂ 0.01). Hepatic Ppara gene expression was upregulated in female offspring only (1.51-fold, p ˂ 0.05) and was restored in the female offspring by folate treatment (p ˂ 0.05). We observed an increase in hepatic Lcn2 and Tmcc2 expression in female offspring born to male rats exposed to an unhealthy diet during spermatogenesis before mating (p ˂ 0.05 vs controls). Folate treatment of the corresponding dams during pregnancy abolished this effect (p ˂ 0.05). Analysis of DNA methylation levels of CpG islands in the Ppara, Lcn2 and Tmcc2 promoter regions revealed that the paternal unhealthy diet induced alterations in the methylation pattern. These patterns were also affected by folate treatment. Total liver DNA methylation was increased by 1.52-fold in female offspring born to male rats on an unhealthy diet prior to mating (p ˂ 0.05). This effect was abolished by folate treatment during pregnancy (p ˂ 0.05 vs the offspring of HFSSD-fed male rats).


Folate treatment of pregnant dams restores effects on female offspring’s glucose metabolism induced by pre-conception male founder HFSSD.


Glucose tolerance High-fat-sucrose-salt diet Maternal folate treatment Paternal programming 



Per cent 5-methylcytosine


High-fat, high-sucrose and high-salt diet




Contribution statement

BH designed the study. JL, MT, XLZ, QZ, OT, JG, MG, AH, CR, X-NP, G-YS, Y-PL and GL generated and analysed the data. JL, OT, BH, Y-PL, AH and CR interpreted the data and wrote the manuscript. All the authors revised the manuscript for intellectual content and approved its final version to be published. BH is the guarantor of this work.


This project has been funded in whole or in part with funds from the National Natural Science Foundation of China (Grant No. 81300557), Hunan Province Science and Technology Plan (grant no. 2014SK3003) and the Programme for Excellent Talents of Hunan Normal University (grant no. ET14106).

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Supplementary material

125_2018_4635_MOESM1_ESM.pdf (227 kb)
ESM (PDF 226 kb)
125_2018_4635_MOESM2_ESM.xlsx (16 kb)
ESM 2 (XLSX 16 kb)
125_2018_4635_MOESM3_ESM.xlsx (43 kb)
ESM 3 (XLSX 43 kb)
125_2018_4635_MOESM4_ESM.xlsx (32 kb)
ESM 4 (XLSX 31 kb)
125_2018_4635_MOESM5_ESM.xlsx (90 kb)
ESM 5 (XLSX 89 kb)
125_2018_4635_MOESM6_ESM.xlsx (133 kb)
ESM 6 (XLSX 132 kb)


  1. 1.
    Reichetzeder C, Dwi Putra SE, Li J, Hocher B (2016) Developmental origins of disease - crisis precipitates change. Cell Physiol Biochem 39:919–938CrossRefPubMedGoogle Scholar
  2. 2.
    Reichetzeder C, Dwi Putra SE, Pfab T et al (2016) Increased global placental DNA methylation levels are associated with gestational diabetes. Clin Epigenetics 8:82CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Novakovic B, Saffery R (2010) DNA methylation profiling highlights the unique nature of the human placental epigenome. Epigenomics 2:627–638CrossRefPubMedGoogle Scholar
  4. 4.
    Hocher B, Haumann H, Rahnenführer J et al (2016) Maternal eNOS deficiency determines a fatty liver phenotype of the offspring in a sex dependent manner. Epigenetics 11:539–552CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Thompson RF, Fazzari MJ, Niu H et al (2010) Experimental intrauterine growth restriction induces alterations in DNA methylation and gene expression in pancreatic islets of rats. J Biol Chem 285:15111–15118CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Woods LL, Weeks DA, Rasch R (2004) Programming of adult blood pressure by maternal protein restriction: role of nephrogenesis. Kidney Int 65:1339–1348CrossRefPubMedGoogle Scholar
  7. 7.
    Barker DJ, Gluckman PD, Godfrey KM et al (1993) Fetal nutrition and cardiovascular disease in adult life. Lancet 341:938–941CrossRefPubMedGoogle Scholar
  8. 8.
    Reichetzeder C, Chen H, Föller M et al (2014) Maternal vitamin D deficiency and fetal programming--lessons learned from humans and mice. Kidney Blood Press Res 39:315–329CrossRefPubMedGoogle Scholar
  9. 9.
    Thone-Reineke C, Kalk P, Dorn M et al (2006) High-protein nutrition during pregnancy and lactation programs blood pressure, food efficiency, and body weight of the offspring in a sex-dependent manner. Am J Phys Regul Integr Comp Phys 291:R1025–R1030Google Scholar
  10. 10.
    Kajantie E, Dunkel L, Turpeinen U et al (2003) Placental 11β-hydroxysteroid dehydrogenase-2 and fetal cortisol/cortisone shuttle in small preterm infants. J Clin Endocrinol Metab 88:493–500CrossRefPubMedGoogle Scholar
  11. 11.
    Li J, Lu YP, Reichetzeder C et al (2016) Maternal PCaaC38:6 is associated with preterm birth - a risk factor for early and late adverse outcome of the offspring. Kidney Blood Press Res 41:250–257CrossRefPubMedGoogle Scholar
  12. 12.
    Hocher B, Slowinski T, Stolze T et al (2000) Association of maternal G protein beta3 subunit 825T allele with low birthweight. Lancet 355:1241–1242CrossRefPubMedGoogle Scholar
  13. 13.
    McPherson NO, Fullston T, Aitken RJ, Lane M (2014) Paternal obesity, interventions, and mechanistic pathways to impaired health in offspring. Ann Nutr Metab 64:231–238CrossRefPubMedGoogle Scholar
  14. 14.
    Li J, Tsuprykov O, Yang X, Hocher B (2016) Paternal programming of offspring cardiometabolic diseases in later life. J Hypertens 34:2111–2126CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Chen Y-P, Xiao X-M, Li J et al (2012) Paternal body mass index (BMI) is associated with offspring intrauterine growth in a gender dependent manner. PLoS One 7:e36329CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Ng S-F, Lin RCY, Laybutt DR et al (2010) Chronic high-fat diet in fathers programs β-cell dysfunction in female rat offspring. Nature 467:963–966CrossRefPubMedGoogle Scholar
  17. 17.
    McPherson NO, Lane M, Sandeman L et al (2017) An exercise-only intervention in obese fathers restores glucose and insulin regulation in conjunction with the rescue of pancreatic islet cell morphology and microRNA expression in male offspring. Nutrients 9:122CrossRefPubMedCentralGoogle Scholar
  18. 18.
    McPherson NO, Owens JA, Fullston T, Lane M (2015) Preconception diet or exercise intervention in obese fathers normalizes sperm microRNA profile and metabolic syndrome in female offspring. Am J Physiol Endocrinol Metab 308:E805–E821CrossRefPubMedGoogle Scholar
  19. 19.
    Lillycrop KA, Phillips ES, Jackson AA et al (2005) Dietary protein restriction of pregnant rats induces and folic acid supplementation prevents epigenetic modification of hepatic gene expression in the offspring. J Nutr 135:1382–1386CrossRefPubMedGoogle Scholar
  20. 20.
    Torrens C, Brawley L, Anthony FW et al (2006) Folate supplementation during pregnancy improves offspring cardiovascular dysfunction induced by protein restriction. Hypertens Dallas Tex 1979 47:982–987Google Scholar
  21. 21.
    Chen Q, Yan M, Cao Z et al (2016) Sperm tsRNAs contribute to intergenerational inheritance of an acquired metabolic disorder. Science 351:397–400CrossRefPubMedGoogle Scholar
  22. 22.
    Waldron D (2016) Non-coding RNA: inheritance of diet-induced metabolic changes via tsRNAs. Nat Rev Genet 17:128PubMedGoogle Scholar
  23. 23.
    Yan M, Zhai Q (2016) Sperm tsRNAs and acquired metabolic disorders. J Endocrinol 230:F13–F18CrossRefPubMedGoogle Scholar
  24. 24.
    Shin JH, Shiota K (1999) Folic acid supplementation of pregnant mice suppresses heat-induced neural tube defects in the offspring. J Nutr 129:2070–2073CrossRefPubMedGoogle Scholar
  25. 25.
    Zhao M, Chen Y-H, Chen X et al (2014) Folic acid supplementation during pregnancy protects against lipopolysaccharide-induced neural tube defects in mice. Toxicol Lett 224:201–208CrossRefPubMedGoogle Scholar
  26. 26.
    Wang X, Tang D, Shen P et al (2017) Analysis of DNA methylation in chondrocytes in rats with knee osteoarthritis. BMC Musculoskelet Disord 18:377CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Paul DS, Guilhamon P, Karpathakis A et al (2014) Assessment of RainDrop BS-seq as a method for large-scale, targeted bisulfite sequencing. Epigenetics 9:678–684CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Feng H, Conneely KN, Wu H (2014) A Bayesian hierarchical model to detect differentially methylated loci from single nucleotide resolution sequencing data. Nucleic Acids Res 42:e69CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Terashima M, Barbour S, Ren J et al (2015) Effect of high fat diet on paternal sperm histone distribution and male offspring liver gene expression. Epigenetics 10:861–871CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Ornellas F, Souza-Mello V, Mandarim-de-Lacerda CA, Aguila MB (2015) Programming of obesity and comorbidities in the progeny: lessons from a model of diet-induced obese parents. PLoS One 10:e0124737CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Ng S-F, Lin RCY, Maloney CA et al (2014) Paternal high-fat diet consumption induces common changes in the transcriptomes of retroperitoneal adipose and pancreatic islet tissues in female rat offspring. FASEB J 28:1830–1841CrossRefPubMedGoogle Scholar
  32. 32.
    Carone BR, Fauquier L, Habib N et al (2010) Paternally induced transgenerational environmental reprogramming of metabolic gene expression in mammals. Cell 143:1084–1096CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Gabory A, Roseboom TJ, Moore T et al (2013) Placental contribution to the origins of sexual dimorphism in health and diseases: sex chromosomes and epigenetics. Biol Sex Differ 4:5CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Oster M, Trakooljul N, Reyer H et al (2017) Sex-specific muscular maturation responses following prenatal exposure to methylation-related micronutrients in pigs. Nutrients 9:74CrossRefPubMedCentralGoogle Scholar
  35. 35.
    Rees WD, Hay SM (2014) Lipocalin-2 (Lcn2) expression is mediated by maternal nutrition during the development of the fetal liver. Genes Nutr 9:380CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Joubert BR, den Dekker HT, Felix JF et al (2016) Maternal plasma folate impacts differential DNA methylation in an epigenome-wide meta-analysis of newborns. Nat Commun 7:10577CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Beckett EL, Veysey M, Lucock M (2017) Folate and microRNA: bidirectional interactions. Clin Chim Acta Int J Clin Chem 474:60–66CrossRefGoogle Scholar
  38. 38.
    Skinner MK (2010) Metabolic disorders: fathers’ nutritional legacy. Nature 467:922–923CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Dwi Putra SE, Reichetzeder C, Meixner M et al (2017) DNA methylation of the glucocorticoid receptor gene promoter in the placenta is associated with blood pressure regulation in human pregnancy. J Hypertens 35:2276–2286CrossRefPubMedGoogle Scholar
  40. 40.
    Hocher B, Adamski J (2017) Metabolomics for clinical use and research in chronic kidney disease. Nat Rev Nephrol 13:269–284CrossRefPubMedGoogle Scholar
  41. 41.
    Tsuprykov O, Ando R, Reichetzeder C et al (2016) The dipeptidyl peptidase inhibitor linagliptin and the angiotensin II receptor blocker telmisartan show renal benefit by different pathways in rats with 5/6 nephrectomy. Kidney Int 89:1049–1061CrossRefPubMedGoogle Scholar
  42. 42.
    de Sousa Rodrigues ME, Bekhbat M, Houser MC et al (2017) Chronic psychological stress and high-fat high-fructose diet disrupt metabolic and inflammatory gene networks in the brain, liver, and gut and promote behavioral deficits in mice. Brain Behav Immun 59:158–172CrossRefPubMedGoogle Scholar
  43. 43.
    Asimakopoulou A, Weiskirchen S, Weiskirchen R (2016) Lipocalin 2 (LCN2) expression in hepatic malfunction and therapy. Front Physiol 7:430CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Abdelmegeed MA, Choi Y, Godlewski G et al (2017) Cytochrome P450-2E1 promotes fast food-mediated hepatic fibrosis. Sci Rep 7:39764CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Borengasser SJ, Kang P, Faske J et al (2014) High fat diet and in utero exposure to maternal obesity disrupts circadian rhythm and leads to metabolic programming of liver in rat offspring. PLoS One 9:e84209CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Williams L, Seki Y, Vuguin PM, Charron MJ (2014) Animal models of in utero exposure to a high fat diet: a review. Biochim Biophys Acta 1842:507–519CrossRefPubMedGoogle Scholar
  47. 47.
    Laubach ZM, Perng W, Dolinoy DC et al (2018) Epigenetics and the maintenance of developmental plasticity: extending the signalling theory framework. Biol Rev Camb Philos Soc.
  48. 48.
    Lövkvist C, Dodd IB, Sneppen K, Haerter JO (2016) DNA methylation in human epigenomes depends on local topology of CpG sites. Nucleic Acids Res 44:5123–5132CrossRefPubMedPubMedCentralGoogle Scholar
  49. 49.
    de Castro BT, Ingerslev LR, Alm PS et al (2016) High-fat diet reprograms the epigenome of rat spermatozoa and transgenerationally affects metabolism of the offspring. Mol Metab 5:184–197CrossRefGoogle Scholar
  50. 50.
    Gapp K, Jawaid A, Sarkies P et al (2014) Implication of sperm RNAs in transgenerational inheritance of the effects of early trauma in mice. Nat Neurosci 17:667–669CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Jian Li
    • 1
  • Yong-Ping Lu
    • 2
    • 3
  • Oleg Tsuprykov
    • 2
    • 4
  • Ahmed A. Hasan
    • 2
    • 5
  • Christoph Reichetzeder
    • 2
  • Mei Tian
    • 1
  • Xiao Li Zhang
    • 1
  • Qin Zhang
    • 1
  • Guo-Ying Sun
    • 1
  • Jingli Guo
    • 2
    • 6
  • Mohamed M. S. Gaballa
    • 2
    • 7
  • Xiao-Ning Peng
    • 1
  • Ge Lin
    • 8
    • 9
    • 10
  • Berthold Hocher
    • 1
    • 2
    Email author
  1. 1.Key Laboratory of Study and Discovery of Small Targeted Molecules of Hunan Province, School of MedicineHunan Normal UniversityChangshaChina
  2. 2.Institute of Nutritional ScienceUniversity of PotsdamNuthetalGermany
  3. 3.Department of Nephrology, Charité - Universitätsmedizin BerlinBerlinGermany
  4. 4.Institute for Laboratory Medicine, IFLBBerlinGermany
  5. 5.Department of Biochemistry, Faculty of PharmacyZagazig UniversityZagazigEgypt
  6. 6.Center for Cardiovascular Research, Charité - Universitätsmedizin BerlinBerlinGermany
  7. 7.Faculty of Veterinary MedicineBenha UniversityToukhEgypt
  8. 8.Institute of Reproductive and Stem Cell Engineering, College of Basic of MedicineCentral South UniversityChangshaChina
  9. 9.Reproductive and Genetic Hospital of CITIC-XiangyaChangshaChina
  10. 10.Key Laboratory of Reproductive and Stem Cell EngineeringNational Health and Family Planning CommissionChangshaChina

Personalised recommendations