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. 2020 Mar 13;21(6):1981.
doi: 10.3390/ijms21061981.

Maternal N-Acetyl Cysteine Intake Improved Glucose Tolerance in Obese Mice Offspring

Michal Michlin  1 , Lital Argaev-Frenkel  1 , Liza Weinstein-Fudim  2 , Asher Ornoy  2   3 , Tovit Rosenzweig  1
Affiliations

Maternal N-Acetyl Cysteine Intake Improved Glucose Tolerance in Obese Mice Offspring

Michal Michlin et al. Int J Mol Sci. .

Abstract

Exposure to certain environmental factors during the early stages of development was found to affect health in adulthood. Among other environmental factors, oxidative stress has been suggested to be involved in fetal programming, leading to elevated risk for metabolic disorders, including type 2 diabetes; however, the possibility that antioxidant consumption during early life may affect the development of diabetes has scarcely been studied. The aim of this study was to investigate the effects of N-acetyl-l-cysteine (NAC) given during pregnancy and lactation on the susceptibility of offspring to develop glucose intolerance at adulthood. C57bl6/J mice were given NAC during pregnancy and lactation. High fat diet (HFD) was given to offspring at an age of 6 weeks for an additional 9 weeks, till the end of the study. Isolated islets of NAC-treated offspring (6 weeks old, before HFD feeding) had an increased efficacy of glucose-stimulated insulin secretion and a higher resistance to oxidative damage. Following HFD feeding, glucose tolerance and insulin sensitivity of NAC-treated offspring were improved. In addition, islet diameter was lower in male offspring of NAC-treated mice compared to their HFD-fed littermates. NAC consumption during early life improves glucose tolerance in adulthood in mice.

Keywords: N-acetyl-l-cysteine; glucose intolerance; high fat diet; metabolic programming; offspring; type 2 diabetes.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
N-acetyl cysteine (NAC) improved glucose tolerance in high fat diet (HFD)-fed offspring of NAC-treated male mice. NAC was given to female mice during pregnancy and lactation. HFD was given to offspring from the age of 6 weeks. Body weight of female (A) and male (B) offspring was measured every week. Glucose tolerance test (GTT) was performed at age of 14 weeks in female (C) and male (D) offspring as described in Materials and Methods. The results are presented as mean ± SE, * p < 0.05. ** p < 0.005, *** p < 0.0005 compared to C/HFD mice, by one-way (A,C) or two-way (C,D) Anova. (E) Area under curve was calculated using GraphPad Prism 8. C/STD: STD-fed offspring of control mice, C/HFD: HFD-fed offspring of control mice, NAC/HFD: HFD-fed offspring of NAC-treated mice. The results are presented as mean ± SE (n = 8), **** p < 0.0001, # p < 0.05 and # p < 0.05, #### p < 0.0001 between genders, by two-way Anova followed by Tukey’s post-test.
Figure 2
Figure 2
NAC improved insulin sensitivity in HFD-fed offspring of NAC-treated male mice. NAC was given to female mice during pregnancy and lactation. HFD was given to offspring from the age of 6 weeks. Insulin tolerance test (ITT) was performed at age of 15 weeks in female (A) and male (B) offspring as described in Methods. The results are presented as mean ± SE (n = 8), * p < 0.05, ** p < 0.005, *** p < 0.0005 by Student’s t-test, compared to C/HFD mice. Relative values are presented in (C) and (D). Fasting serum insulin was measured (n ≥ 8) at age of 16 weeks (E). H&E staining of pancreas was performed (5 mice, 3 section for each pancreas), and islet diameter (F) and number of islets per section (G) were measured. C/STD: STD-fed offspring of control mice, C/HFD: HFD-fed offspring of control mice, NAC/HFD: HFD-fed offspring of NAC-treated mice. The results are presented as mean ± SE, ** p < 0.005, *** p < 0.0005, **** p < 0.0005 and ## p < 0.005, ### p < 0.0005, between genders, in two-way Anova followed by Tukey’s post-test.
Figure 3
Figure 3
NAC improved the transmission of insulin signaling in HFD-fed offspring of NAC-treated male mice. NAC was given to females during pregnancy and lactation. HFD was given to offspring from the age of 6 weeks. Soleus muscle and liver of male (A,B, respectively) and soleus muscle and liver of female (C,D, respectively) offspring were removed at the age of 16 weeks as described in the Methods (n = 5). Western blot analysis was performed on protein lysates of muscle and liver using specific antibodies. C/STD: STD-fed offspring of control mice, C/HFD: HFD-fed offspring of control mice, NAC/HFD: HFD-fed offspring of NAC-treated mice.
Figure 4
Figure 4
NAC reduced severity of hepatic steatosis and adipose tissue inflammation in HFD-fed offspring male mice. NAC was given to females during pregnancy and lactation. HFD was given to offspring from the age of 6 weeks. Mice were killed at age 16 weeks and H&E staining of liver (A) and adipose tissue (E) was performed. Severity of steatosis was graded using steatosis score, as described in Methods (B). Visceral adipose tissue weight was measured (C) and adipocyte diameter was measured (D). C/STD: STD-fed offspring of control mice, C/HFD: HFD-fed offspring of control mice, NAC/HFD: HFD-fed offspring of NAC-treated mice. The results are presented as mean ± SE (n ≥ 4), ** p < 0.005, **** p < 0.005 in one-way Anova followed by Tukey’s post-test.
Figure 5
Figure 5
Effect of early life administration of NAC on markers of oxidative stress in HFD-fed mice. NAC was given to females during pregnancy and lactation. HFD was given to offspring from the age of 6 weeks. Mice were killed at age 16 weeks and serum (A) and liver (B) thiobarbituric acid reactive substrate (TBARS) were measured. (C) Protein carbonylation was measured in liver, as describes in Methods. The results are presented as mean ± SE (n = 8), * p < 0.05, ** p < 0.005, **** p < 0.0.0005 in one-way Anova followed by Tukey’s post-test. ### p < 0.0005 in two way Anova followed by Bonferroni’s post-test.
Figure 6
Figure 6
NAC improved pancreatic islet function in young offspring. NAC was given to females during pregnancy and lactation. Offspring were killed at age of 6 weeks, H&E staining of pancreas was performed, and number of islets per section (A) and islet diameter (B) were measured (5 mice, 3 sections for each pancreas). In additional set of 6 weeks old offspring, pancreatic islets were isolated and glucose stimulated insulin secretion (GSIS) was performed as described in Methods (C,D). The results are presented as mean ± SE (n = 6, 3 internal replicates for each experiment), * p < 0.05, *** p < 0.0005 in one-way Anova followed by Tukey’s post-test.
Figure 7
Figure 7
NAC increased the resistance of pancreatic islets against H2O2-induced oxidative stress. NAC was given to females during pregnancy and lactation. Pancreatic islets of offspring were isolated at age of 6 weeks. (A) Isolated pancreatic islets were treated with different doses of H2O2, and stained with propidium iodide, as described in Methods. A total of 3 representing micrographs are presented. The number of dead cells/islets was quantified in female (B) and male (C) mice. (D) mRNA was isolated from islets of 6 weeks old mice, and expression of selected genes was measured by RTPCR as described in Methods. Results were normalized to the expression of the housekeeping gene, RPS29 and are presented as the mean ± SEM. (E) Immunohistochemistry for the expression of SOD and Catalase was performed in pancreas of 6 weeks old male mice, and the expression level was blindly scored. The results are presented as mean ± SEM (n = 5, 3 replicates for each experiment), * p < 0.05, ** p < 0.005 and *** p < 0.0005 in one-way Anova followed by Tukey’s post-test.
Figure 8
Figure 8
Study design. C57bl6/J mice were given STD with or without NAC supplementation during pregnancy and lactation. Offspring were separated at the age of 3 weeks. Analysis of islet mass and function was performed at age of 6 weeks. STD or HFD were given at age of 6 weeks till the end of the study.
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References

    1. Shaw J.E., Sicree R.A., Zimmet P.Z. Global estimates of the prevalence of diabetes for 2010 and 2030. Diabetes Res. Clin. Pract. 2010;87:4–14. doi: 10.1016/j.diabres.2009.10.007. - DOI - PubMed
    1. Bellou V., Belbasis L., Tzoulaki I., Evangelou E. Risk factors for type 2 diabetes mellitus: An exposure-wide umbrella review of meta-analyses. PLoS ONE. 2018;13:e0194127. doi: 10.1371/journal.pone.0194127. - DOI - PMC - PubMed
    1. Franks P.W., McCarthy M.I. Exposing the exposures responsible for type 2 diabetes and obesity. Science. 2016;354:69–73. doi: 10.1126/science.aaf5094. - DOI - PubMed
    1. Elksnis A., Martinell M., Eriksson O., Espes D. Heterogeneity of metabolic defects in type 2 diabetes and its relation to reactive oxygen species and alterations in beta-cell mass. Front. Physiol. 2019;10:107. doi: 10.3389/fphys.2019.00107. - DOI - PMC - PubMed
    1. Rolo A.P., Teodoro J.S., Palmeira C.M. Role of oxidative stress in the pathogenesis of nonalcoholic steatohepatitis. Free Radic. Biol. Med. 2012;52:59–69. doi: 10.1016/j.freeradbiomed.2011.10.003. - DOI - PubMed

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