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Diabetologia

, Volume 57, Issue 10, pp 2165–2172 | Cite as

Maternal vitamin D deficiency during pregnancy results in insulin resistance in rat offspring, which is associated with inflammation and Iκbα methylation

  • Huaqi Zhang
  • Xia Chu
  • Yifan Huang
  • Gang Li
  • Yuxia Wang
  • Ying LiEmail author
  • Changhao SunEmail author
Article

Abstract

Aims/hypothesis

We aimed to investigate the impact of maternal vitamin D deficiency during pregnancy on insulin resistance in male offspring and examine its mechanism.

Methods

Pregnant Sprague–Dawley rats were maintained on a vitamin-D-free diet with ultraviolet-free light during pregnancy (early-VDD group). Insulin resistance in the male offspring was assessed by HOMA-IR, OGTT and euglycaemic clamp. NEFA, oxidative stress and inflammation levels were estimated as risk factors for insulin resistance. DNA methylation was examined by bisulfate sequencing PCR analysis. Luciferase reporter assay was performed to validate the effect of DNA methylation.

Results

The offspring in the early-VDD group had significantly higher fasting insulin and HOMA-IR levels, markedly reduced glucose tolerance and significantly lower tissue sensitivity to exogenous insulin at 16 weeks (all p < 0.05) compared with control offspring. Significantly higher serum and liver IL-1β, IL-6, IL-8 and TNF-α concentrations were observed in the offspring of the early-VDD group at 0, 3, 8 and 16 weeks. Expression of hepatic Iκbα (also known as Nfkbia) mRNA and nuclear factor κB inhibitor α (IκBα) protein was persistently lower in the early-VDD offspring at all time points, and their hepatic Iκbα methylation levels at the cytosine phosphate guanine site +331 were significantly higher at 0 and 16 weeks (all p < 0.01). Methylation at Iκbα first exon +331 markedly decreased the luciferase activity (p < 0.05).

Conclusions/interpretation

Maternal vitamin D deficiency during pregnancy results in insulin resistance in the offspring, which is associated with persistently increased inflammation. Persistently decreased Iκbα expression, potentially caused by changes in Iκbα methylation, plays an important role in persistent inflammation.

Keywords

DNA methylation Inflammation Insulin resistance Vitamin D deficiency 

Abbreviations

1,25(OH)D

1,25-Dihydroxyvitamin D

25(OH)D

25-Hydroxyvitamin D

CpG

Cytosine phosphate guanine

Early-VDD

Early-life vitamin D deficiency

FG

Fasting glucose

FIN

Fasting insulin

GIR

Glucose infusion rate

GSH-Px

Glutathione peroxidase

IκBα

NFκB inhibitor α

IKK

NFκB inhibitor kinase

IR

Insulin resistance

MDA

Malonaldehyde

NFκB

Nuclear factor κB

P-IκBα

Phosphorylated NFκB inhibitor α

PTH

Parathyroid hormone

ROS

Reactive oxygen species

SOD

Superoxide dismutase

TG

Triacylglycerol

Notes

Acknowledgements

We thank J. Li (Department of Nutrition and Food Hygiene, Public Health College, Harbin Medical University, Harbin, China) for helping with the revision and L. Y. Gong and J. N. Song (Department of Nutrition and Food Hygiene, Public Health College, Harbin Medical University) for their help with animal feeding and molecular biology experiments.

Funding

This study was supported by the State Key Program of National Natural Science of China (No. 81130049), the National ‘Twelfth Five-Year’ Plan for Science & Technology Support (No. 2012BAI02B02) and the Program for New Century Excellent Talents in University of Ministry of Education of China (grant NCET-10-0148).

Duality of interest

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

Contribution statement

CS and YL designed the study, analysed and interpreted the data and reviewed and edited the manuscript. HZ designed and performed the study, acquired and analysed the data and wrote the draft of the manuscript. XC, YH, GL and YW were responsible for performing the study, data collection and interpretation and drafting the manuscript. All authors approved the final version of the manuscript. CS is responsible for the integrity of the work as a whole.

Supplementary material

125_2014_3316_MOESM1_ESM.pdf (30 kb)
ESM Methods (PDF 30 kb)
125_2014_3316_MOESM2_ESM.pdf (58 kb)
ESM Table 1 (PDF 58 kb)
125_2014_3316_MOESM3_ESM.pdf (29 kb)
ESM Table 2 (PDF 28 kb)
125_2014_3316_MOESM4_ESM.pdf (61 kb)
ESM Fig. 1 (PDF 61 kb)
125_2014_3316_MOESM5_ESM.pdf (108 kb)
ESM Fig. 2 (PDF 108 kb)
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ESM Fig. 3 (PDF 109 kb)
125_2014_3316_MOESM7_ESM.pdf (178 kb)
ESM Fig. 4 (PDF 178 kb)
125_2014_3316_MOESM8_ESM.pdf (144 kb)
ESM Fig. 5 (PDF 143 kb)
125_2014_3316_MOESM9_ESM.pdf (105 kb)
ESM Fig. 6 (PDF 105 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  1. 1.Department of Nutrition and Food Hygiene, Public Health CollegeHarbin Medical UniversityHarbinPeople’s Republic of China
  2. 2.The Second Affiliated Hospital of Harbin Medical UniversityHarbinPeople’s Republic of China
  3. 3.The Third Affiliated Hospital of Harbin Medical UniversityHarbinPeople’s Republic of China

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