Diabetes Metab J.  2016 Dec;40(6):494-508. 10.4093/dmj.2016.40.6.494.

Investigating Susceptibility to Diabetes Using Features of the Adipose Tissue in Response to In Utero Polycyclic Aromatic Hydrocarbons Exposure

• Gato WE
• Hunter DA
• Whitby SL
• Mays CA
• Yau W

• KMID: 2362222
• DOI: http://doi.org/10.4093/dmj.2016.40.6.494

Abstract

BACKGROUND
In recent times, there has been an increase in the incidence of type 2 diabetes mellitus (T2DM) particularly in children. Adipocyte dysfunction provide a critical link between obesity and insulin resistance resulting in diabetes outcome. Further, environmental chemical exposure during early years of life might be a significant contributing factor to the increase in the incidence of T2DM. This study tests the idea that exposure to environmental contaminants (2-aminoanthracene [2AA]) in utero will show effects in the adipose tissue (AT) that signify T2DM vulnerability. 2AA is a polycyclic aromatic hydrocarbon found in a variety of products.
METHODS
To accomplish the study objective, pregnant dams were fed various amounts of 2AA adulterated diets from gestation through postnatal period. The neonates and older offspring were analyzed for diabetic-like genes in the ATs and analysis of serum glucose. Furthermore, weight monitoring, histopathology and immunohistochemical (IHC) staining for CD68 in AT, adipocyte size determination and adiponectin amounts in serum were undertaken.
RESULTS
Up-regulation of adiponectin and interleukin-6 genes were noted in the pups and older rats. Combination of intrauterine 2AA toxicity with moderate high fat diet exhibited gene expression patterns similar to those of the neonates. Elevated serum glucose levels were noted in treated groups. IHC of the AT indicated no significant malformations; however, CD68+ cells were greater in the animals treated to 2AA. Similarly, mean sizes of the adipocytes were larger in treated and combined 2AA and moderate high fat animals. Adiponectin was reduced in 2AA groups.
CONCLUSION
From the preceding, it appears intrauterine 2AA disturbance, when combined with excess fat accumulation will lead to greater risk for the diabetic condition.

Keyword

Adipocytes; Adiponectin; Diabetes; Gene expression; Interleukin-6; Serum glucose; 2-Anthramine

MeSH Terms

Adipocytes
Adiponectin
Adipose Tissue*
Animals
Blood Glucose
Child
Diabetes Mellitus, Type 2
Diet
Diet, High-Fat
Gene Expression
Humans
Incidence
Infant, Newborn
Insulin Resistance
Interleukin-6
Obesity
Polycyclic Hydrocarbons, Aromatic*
Pregnancy
Rats
Up-Regulation
Adiponectin
Interleukin-6
Polycyclic Hydrocarbons, Aromatic

Figure

• 

  Fig. 1 Mean weights, g, (n=12 to 15) of each treatment group over 14 weeks. Broadly, progeny were exposed to 0 mg/kg 2-aminoanthracene (2AA) diet (control), 50 mg/kg 2AA diet (low dose), and 100 mg/kg 2AA diet (high dose) in utero. Time seem to have an effect on weight gain, no statistical differences were noted with regards to 2AA treatment.

• 

  Fig. 2 Histologic characterization (H&E stain, 100 µm) of adipose tissue of Sprague Dawley rats exposed to (A) 0 mg/kg- (control [C] pup), (B) 50 mg/kg- (low dose [LD] pup), and (C) 100 mg/kg-2-aminoanthracene (high dose [HD] pup) from gestation through of the postnatal period (H&E stain). Older pups were fed moderate high and regular diet for 6 weeks. Select images include: (D) control female regular diet (CFR); (E) control male regular diet (CMR); (F) control high fat diet (CFH); (G) control male high fat diet (CMH); (H) low dose male high fat (LMH); and (I) high dose male high fat diet (HMH). Small to moderate numbers of macrophages are scattered between adipocytes in all three groups of young pups. Fewer numbers of macrophages are present in all groups of older pups.

• 

  Fig. 3 Numbers of CD68 positive cells (n=6 to 8) in adipose tissue of Sprague Dawley rats. (A) Intrauterine treatment to 2-aminoanthracene groups included: control (C) pup (0 mg/kg diet), low dose (LD) pup (50 mg/kg diet), and high dose (HD) pup (100 mg/kg diet). (B) In utero exposure was combined with dietary ingestions of regular diet (5M30 rodent diet; PMI Nutrition International) and high fat diet (Adjusted Fat Diet TD.96132) for 6 weeks 3 months postwean. Though the treated exposed rats indicated higher CD68 positive values, statistical inference showed they were not significant. CMH, control male high fat; CMR, control male regular diet; LMH, low dose male high fat; CFR, control female regular diet; LFR, low dose female regular diet; HFR, high dose female regular diet; HMH, high dose high fat.

• 

  Fig. 4 Mean size of adipocytes (pixels) (n=5 to 6). Significant increment in the adipocyte size observed for the LMH and HMH animals. CMH, control male high fat; CMR, control male regular diet; LMH, low dose male high fat; CFR, control female regular diet; LFR, low dose female regular diet; HFR, high dose female regular diet; HMH, high dose high fat. aP<0.05.

• 

  Fig. 5 Mean serum glucose concentration (mg/dL) of neonates exposed to 2-aminoanthracene (2AA) in utero. (A) Dams ingested 0 mg/kg (control [C]); 50 mg/kg (low dose [LD]); and 100 mg/kg (high dose [HD])-2AA from gestation through 2 weeks postnatal period; (B) control male high fat (CMH), low dose male high fat (LMH), and high dose high fat (HMH); (C) control female regular diet (CFR), low dose female regular diet (LFR), and high dose female regular diet (HFR); (D) control male regular diet (CMR), low dose male regular diet (LMR), and high dose male regular diet (HMR). Significant differences in serum glucose concentration were noted as aP<0.05, bP<0.01.

• 

  Fig. 6 Relative expression (ΔCq) of adipose tissue genes involved in diabetic-related conditions. In utero exposure to 2-aminoanthracene (2AA) were 0 mg/kg 2AA diet (control [C]), 50 mg/kg diet (low dose [LD]), and 100 mg/kg diet (high dose [HD]) from gestation through 14 days postnatal. IL-6, interleukin-6; TNF-α, tumor necrosis factor α. Significant changes in differential gene expression is noted as aP<0.05, bP<0.01.

• 

  Fig. 7 Relative expression (ΔCq) of adipokine and cytokines involved in diabetic-related conditions. (A) In utero exposure of 0 mg/kg 2-aminoanthracene diet (control), 50 mg/kg diet (low dose), and 100 mg/kg diet (high dose) from gestation through 14 days postnatal was combined with dietary ingestions of regular diet (5M30 rodent diet; PMI Nutrition International) and high fat diet (Adjusted Fat Diet TD.96132) for 6 weeks 3 months postwean. (A) Control male regular diet (CMR), low dose male regular diet (LMR), and high dose male regular diet (HMR). (B) Control female regular diet (CFR), low dose female regular diet (LFR), and high dose female regular diet (HFR). (C) Control male high fat (CMH), low dose male high fat (LMH), and high dose high fat (HMH). IL-6, interleukin-6; TNF-α, tumor necrosis factor α. Significant differences (aP<0.05, bP<0.01) in adiponectin and IL-6 mRNA expression were noted.

• 

  Fig. 8 (A-D) Serum adiponectin levels in rats exposed to 2-aminoanthracene (2AA) in utero and moderate high fat 3 months postwean. Changes in adiponectin amounts were significant (P<0.01) in neonatal rats but not significantly altered in the regular dietary female animals. Both the male regular and high fat dietary group showed significant (P<0.05) reduction of adiponectin in 2AA treated rats. C, control; LD, low dose; HD, high dose; CMH, control male high fat; LMH, low dose male high fat; HMH, high dose high fat; CFR, control female regular diet; LFR, low dose female regular diet; HFR, high dose female regular diet; CMR, control male regular diet; LMR, low dose male regular diet; HMR, high dose male regular diet. aP<0.05, bP<0.01.

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