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Maternal obesity and high-fat diet program offspring metabolic syndrome

Published:March 17, 2014DOI:https://doi.org/10.1016/j.ajog.2014.03.025

      Objective

      We determined the potential programming effects of maternal obesity and high-fat (HF) diet during pregnancy and/or lactation on offspring metabolic syndrome.

      Study Design

      A rat model of maternal obesity was created using an HF diet prior to and throughout pregnancy and lactation. At birth, pups were cross-fostered, thereby generating 4 paradigms of maternal diets during pregnancy/lactation: (1) control (Con) diet during pregnancy and lactation (Con/Con), (2) HF during pregnancy and lactation (HF/HF), (3) HF during pregnancy alone (HF/Con), and (4) HF during lactation alone (Con/HF).

      Results

      Maternal phenotype during pregnancy and the end of lactation evidenced markedly elevated body fat and plasma corticosterone levels in HF dams. In the offspring, the maternal HF diet during pregnancy alone programmed increased offspring adiposity, although with normal body weight, whereas the maternal HF diet during lactation increased both body weight and adiposity. Metabolic disturbances, particularly that of hyperglycemia, were apparent in all groups exposed to the maternal HF diet (during pregnancy and/or lactation), although differences were apparent in the manifestation of insulin resistant vs insulin-deficient phenotypes. Elevated systolic blood pressure was manifest in all groups, implying that exposure to an obese/HF environment is disadvantageous for offspring health, regardless of pregnancy or lactation periods. Nonetheless, the underlying mechanism may differ because offspring that experienced in utero HF exposure had increased corticosterone levels.

      Conclusion

      Maternal obesity/HF diet has a marked impact on offspring body composition and the risk of metabolic syndrome was dependent on the period of exposure during pregnancy and/or lactation.

      Key words

      An epidemic of metabolic syndrome is well recognized within the United States. Currently 65% of adult Americans are overweight and 30% obese, although a marked increase in obesity is apparent from childhood through adolescence.
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      Gestational weight gain in relation to offspring body mass index and obesity from infancy through adulthood.
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      Thus, maternal obesity may affect breast milk–induced newborn programming, independent of the maternal obesity effects on fetal programming.
      In view of the potential programming effects of maternal obesity during both pregnancy and lactation, we sought to examine the effects of each of these periods on programmed metabolic syndrome in rats. We hypothesized that offspring exposed to maternal obesity during both pregnancy and lactation would be more obese and exhibit a greater degree of metabolic abnormalities.

      Materials and Methods

      Maternal diet and studies

      A rat model of maternal obesity was created using a HF diet prior to and through pregnancy and lactation was utilized. Studies were approved by the Animal Research Committee of Harbor-UCLA Medical Research and Education Institute and were in accordance with the American Association for Accreditation of Laboratory Care and National Institutes of Health guidelines.
      Sprague Dawley rats (Charles River Laboratories, Inc, Hollister, CA) were housed in a facility with constant temperature and humidity and on a controlled 12 hour light–12 hour dark cycle. Beginning as weanlings, female rats were fed a HF (60% k/cal fat; Purified Diet 58Y1, New Brunswick, NJ; n = 16) or control (Con; 10% k/cal fat, Purified Diet 58Y2; n = 16) diet. The nutrient composition is given in Table 1. At 11 weeks of age, the rats were mated and continued on their respective diets during pregnancy and lactation.
      Table 1Nutrient composition of diets
      VariablePurified diet 58Y2, 10% k/cal fatPurified diet 58Y1, 60% k/cal fat
      Nutrients, %
       Carbohydrates67.425.9
       Protein17.323.1
       Fat4.334.9
       Lard1.931.7
       Soybean oil2.43.2
      Fat type, %
       Saturated2537
       Monounsaturated3546
       Polyunsaturated4017
      Nutrient values are percentage per 100 g food, and fat type is percentage of total fat.
      Desai. Maternal obesity programs offspring metabolic syndrome. Am J Obstet Gynecol 2014.
      Maternal body weights and their food intake were recorded daily. In addition, maternal blood was obtained via a tail bleed at term (gestational age embryonic day [e] 20) and at the end of lactation (21 days postpartum) for glucose, lipid, and hormonal analysis (details are given in the following text). The food was removed 1 hour prior to the blood sampling. Furthermore, at the end of lactation, dams underwent a noninvasive dual-energy x-ray absorptiometry (DEXA) scan (details are given in the following text) for evaluating percentage body fat and lean body mass.

      The offspring

      At birth, pups were culled to 8 per litter (4 males and 4 females) to normalize rearing and were cross-fostered, thereby generating 4 paradigms of maternal diets during pregnancy/lactation: (1) Con diet during pregnancy and lactation (Con/Con), (2), HF during pregnancy and lactation (HF/HF), (3) HF during pregnancy alone (HF/Con), and (4) HF during lactation alone (Con/HF). At 3 weeks of age, the offspring in each of the 4 groups were housed individually and 2 males and 2 females per litter were weaned to a normal-fat diet (10% k/cal).

      Body weights and food intake

      Each litter from the 4 groups was weighed weekly, and the weight of an individual pup was calculated from it (ie, litter weight per number of pups). The first weight was recorded at 1 day of age, and subsequent weights were taken at 7, 14, and 21 days of age. Thereafter the body weight and food intake were monitored weekly on an individual basis.

      Body composition

      At 3 and 24 weeks of age, offspring of both sexes underwent noninvasive DEXA scanning using the DEXA system with a software program for small animals (QDR 4500A; Hologic, Bedford, MA). An in vivo scan of whole-body composition was obtained, including the lean and fat tissue mass, the total mass, and the percentage body fat measurements.

      Blood pressure

      Blood pressure was determined in conscious 8 week old male and female offspring using a noninvasive tail-cuff sphygmomanometry (ML125 NIPB system; AD Instruments, Colorado Springs, CO) method. Several cuff sizes were used, depending on the weight of the animal. To circumvent the potential problem of restrain-induced stress, the animals were acclimatized for at least 1 week with placement in the restraint.

      Glucose tolerance test

      At 6 and 24 weeks of age, offspring of both sexes underwent a glucose tolerance test (GTT) as follows: after an overnight fast, D-glucose (1 mg/g body weight) was injected intraperitoneally in conscious rats. Blood was taken via a tail bleed prior to (time 0) and 15, 30, 60, 120, and 180 minutes after glucose injection.

      Blood collection

      One day after birth (approximately 24 hours after birth), the excess pups (4 pups per litter) from Con (n = 16) and HF (n = 16) groups were decapitated and blood was pooled from pups from the same litter. At ages 3 and 24 weeks, 1 male and 1 female from each litter (n = 8 per group) were fasted overnight, and blood was collected via cardiac puncture in heparinized tubes for plasma analysis.

      Plasma analysis

      Plasma insulin, leptin, and coritcosterone levels were measured using rat-specific commercial radioimmunoassay kits (insulin and leptin radioimmunoassay kit; LINCO Research Inc, St Charles, MO; coritcosterone radioimmunoassay kit; Diagnostic Systems Laboratories, Inc, Webster, TX). Plasma lipid levels were measured using reagents from Raichem, Inc (San Diego, CA) and run on an automated Cobas-Mira chemistry analyzer (Roche Diagnostic Systems Inc, Sommerville, NJ). Plasma triglyceride (catalog no. 80008) and cholesterol (catalog no. 80015) concentrations were analyzed using Raichem enzymatic reagents (with control serum level 1 [no. 83082] and control serum level 2 [no. 83083]). Blood glucose was determined using a Hemocue B-glucose analyzer (HemoCue Inc, Mission Viejo, CA).

      Statistical analysis

      For all offspring studies at 3, 6, 8, and 24 weeks of age, 8 males and 8 females from 8 litters were studied per group. Differences between Con and the experimental groups were compared using an unpaired Student t test (1 day old neonate), repeated measures of analysis of variance (body weight and food intake), or analysis of variance with Dunnett's post hoc tests (body composition and plasma hormones/metabolites). At ages 1 day and 3 weeks, combined data for males and females are shown because no sex differences were evident. However, at the age 24 weeks, sex differences justified analyzing the data according to sex. Values are expressed as means ± SE.

      Results

      Maternal dams

      Pregnancy

      Maternal body weight was increased at the initiation of pregnancy as per the experimental model. Both HF and Con dams gained nearly identical amounts of weight during the pregnancy (Figure 1, A). However, there were marked differences immediately following delivery because Con dams experienced a significantly greater weight loss from pregnancy day e20 to postnatal day one (100 ± vs 40 g; Figure 1, B). Among the 4 lactation groups, 3 of the groups demonstrated a similar maternal weight change during lactation (Con/Con, Con/HF, and HF/HF) with a slight increase in maternal weight through postnatal day (p) 10-12 and a slight decrease in maternal weight at the completion of lactation (p20). The HF dams nursing Con pups evidenced a continued decrease in maternal weight throughout the lactation period.
      Figure thumbnail gr1
      Figure 1Maternal body weights during pregnancy and lactation
      A, Daily maternal body weight during pregnancy (e8–e20) of Con (filled circles) and HF dams (open circles). B, Daily maternal body weight during lactation (day 2–20) of Con dams nursing Con pups (filled circles), HF dams nursing HF pups (open circles), Con dams nursing HF pups (filled triangles), and HF dams nursing Con pups (open triangles). Values are mean ± SE of 16 dams per group during pregnancy and 8 dams per group during lactation.
      Con, control; HF, high-fat.
      Desai. Maternal obesity programs offspring metabolic syndrome. Am J Obstet Gynecol 2014.
      When assessed by the percentage of carbohydrate, fat, and protein intake, HF dams consumed a greater percentage of kilocalories via fat and a lower percentage by carbohydrate during both pregnancy and lactation, with nearly identical levels of protein intake as Con dams (Figure 2). The total food intake was similar among HF and Con dams during pregnancy. However, food intake was significantly greater during the terminal portion of lactation among HF dams nursing HF pups and HF dams nursing Con pups (Figure 3).
      Figure thumbnail gr2
      Figure 2Calorie intake during pregnancy and lactation
      A, Daily maternal calorie intake during pregnancy (e8–e20) of Con (filled circles) and HF dams (open circles). B, Daily maternal calorie intake during lactation (day 2–20) of Con dams nursing Con pups (filled circles), HF dams nursing HF pups (open circles), Con dams nursing HF pups (filled triangles), and HF dams nursing Con pups (open triangles). Values are mean ± SE of 16 dams per group during pregnancy and 8 dams per group during lactation.
      Con, control; HF, high-fat.
      Desai. Maternal obesity programs offspring metabolic syndrome. Am J Obstet Gynecol 2014.
      Figure thumbnail gr3
      Figure 3Total maternal calorie intake during pregnancy and lactation
      A, Daily maternal total calorie intake during pregnancy (e8–e20) of Con (filled circles) and HF dams (open circles). B, Daily maternal total calorie intake during lactation (day 2–20) of Con dams nursing Con pups (filled circles), HF dams nursing HF pups (open circles), Con dams nursing HF pups (filled triangles), and HF dams nursing Con pups (open triangles). Values are mean ± SE of 16 dams per group during pregnancy and 8 dams per group during lactation.
      Con, control; HF, high-fat.
      Desai. Maternal obesity programs offspring metabolic syndrome. Am J Obstet Gynecol 2014.
      In conjunction with maternal obesity and a HF maternal diet, the plasma profile of pregnant dams at gestation day e21 included increased plasma cholesterol and coritcosterone, although HF and Con dams had similar plasma leptin, plasma triglyceride, and blood glucose levels (Table 2).
      Table 2Plasma profile of pregnant dams
      VariableConHF
      Plasma triglycerides, mg/dL164 ± 13196 ± 16
      Plasma cholesterol, mg/dL110 ± 5128 ± 3
      P < .05 vs Con.
      Blood glucose, mg/dL57 ± 363 ± 5
      Plasma leptin, ng/mL3.3 ± 0.34.3 ± 0.5
      Plasma corticosterone, ng/mL266 ± 34776 ± 64
      P < .05 vs Con.
      Plasma lipid, glucose, and hormone levels of pregnant dams at term (gestational age e20). Values are mean ± SE of 16 dams per group.
      Con, control; HF, high-fat.
      Desai. Maternal obesity programs offspring metabolic syndrome. Am J Obstet Gynecol 2014.
      a P < .05 vs Con.

      Lactation

      At the completion of lactation, there were marked differences among the dams, dependent on the offspring being nursed (Table 3). HF/HF dams demonstrated markedly increased percentage body fat and reduced lean body mass as compared with all groups. Notably, the HF/Con dams' body fat was significantly less than the HF/HF, consistent with the loss of maternal body weight among HF/Con dams during lactation. Plasma leptin levels reflected the percentage body fat with elevated levels in the HF/HF group. Plasma cholesterol levels, which were elevated among HF dams at the completion of pregnancy, demonstrated a reduction in HF/HF dams as well as HF/Con dams, coinciding with lower levels of plasma triglycerides. Blood glucose levels were similar among all groups, although HF/HF and HF/Con dams demonstrated elevated levels of plasma insulin. Notably, Con/HF dams had markedly elevated levels of plasma corticosterone at the completion of lactation.
      Table 3Plasma profile of lactating dams
      VariableCon dams nursing Con pupsCon dams nursing HF pupsHF dams nursing HF pupsHF dams nursing Con pups
      Body fat, %4.6 ± 0.45.6 ± 0.612.1 ± 1.2
      P < .05 vs Con.
      7.5 ± 0.8
      P < .05 vs Con.
      Lean body mass, %92.9 ± 0.391.5 ± 0.685.4 ± 1.2
      P < .05 vs Con.
      90.0 ± 1.9
      Plasma triglycerides, mg/dL72 ± 960 ± 943 ± 6
      P < .05 vs Con.
      45 ± 3
      P < .05 vs Con.
      Plasma cholesterol, mg/dL95 ± 791 ± 654 ± 5
      P < .05 vs Con.
      65 ± 2
      P < .05 vs Con.
      Blood glucose, mg/dL57 ± 753 ± 668 ± 762 ± 5
      Plasma leptin, ng/mL1.6 ± 0.31.3 ± 0.22.1 ± 0.4
      P < .05 vs Con.
      1.6 ± 0.4
      Plasma insulin, ng/mL0.12 ± 0.020.16 ± 0.020.23 ± 0.05
      P < .05 vs Con.
      0.18 ± 0.02
      P < .05 vs Con.
      Plasma corticosterone, ng/mL247 ± 47588 ± 47
      P < .05 vs Con.
      414 ± 45
      P < .05 vs Con.
      397 ± 45
      P < .05 vs Con.
      Body composition and plasma lipid, glucose, and hormone levels of lactating dams at day 21. Values are mean ± SE of 8 dams per group.
      Con, control; HF, high-fat.
      Desai. Maternal obesity programs offspring metabolic syndrome. Am J Obstet Gynecol 2014.
      a P < .05 vs Con.

      The offspring

      Growth

      Both Con and HF newborns at day p1 had similar body weights (7.3 ± 0.1; 7.4 ± 0.2, respectively). During the nursing period, offspring weight gain diverged into a rapid weight gain group among HF/HF and Con/HF offspring, which was evident in both males and females by 3 weeks of age (Figure 4). Females continued the divergent pattern through 30 weeks of age, whereas males exhibited 3 growth patterns as follows: HF/HF offspring demonstrated a continued acceleration of body weight gain through 30 weeks of age, Con/HF males demonstrated intermediate growth, and HF/Con demonstrating weight gain similar to Con/Con.
      Figure thumbnail gr4
      Figure 4Offspring body weights
      Mean body weights of male and female offspring from 1 to 24 weeks of age in Con (filled circles; pups born to Con dams and nursed by Con dams), HF/HF (open circles; pups born to HF dams and nursed by HF dams), HF/Con (open triangles; pups born to HF dams and nursed by Con dams), and Con/HF (filled triangles; pups born to Con dams and nursed by HF dams). Insets demonstrate body weights of males and females from 1 day to 3 weeks of age. Number of offspring studied per group was 32 males and 32 females (from 8 litters) until 3 weeks of age, after which half the offspring were weaned onto a Con diet. Thereafter the data from 16 males and 16 females (from 8 litters) until 24 weeks of age per group are shown. Values are mean ± SE.
      Con, control; HF, high-fat.
      P < .05 vs Con.
      Desai. Maternal obesity programs offspring metabolic syndrome. Am J Obstet Gynecol 2014.

      Food intake

      Unlike the females, the food intake of male offspring paralleled weight gain, with HF/HF offspring demonstrating the greatest food intake from the end of lactation through 24 weeks of age. Con/HF offspring demonstrated intermediate food intake, slightly greater than Con offspring. Notably, HF/Con offspring demonstrated accelerated food intake through 18 weeks of age, after which food intake normalized to levels of the Con/Con offspring (Figure 5).
      Figure thumbnail gr5
      Figure 5Offspring food intake
      Mean food intake of male and female offspring from 5 to 30 weeks of age in Con (filled circles; pups born to Con dams and nursed by Con dams), HF/HF (open circles; pups born to HF dams and nursed by HF dams), HF/Con (open triangles; pups born to HF dams and nursed by Con dams), and Con/HF (filled triangles; pups born to Con dams and nursed by HF dams). Number of offspring studied per group was 32 males and 32 females (from 8 litters) until 3 weeks of age, after which all offspring were weaned onto a Con diet. Thereafter data from 16 males and 16 females (from 8 litters) until 30 weeks of age per group are shown. Values are mean ± SE.
      Con, control; HF, high-fat.
      P < .05.
      Desai. Maternal obesity programs offspring metabolic syndrome. Am J Obstet Gynecol 2014.

      Body composition

      Consistent with the weight gain, both HF/HF and Con/HF offspring demonstrated significantly increased percentage body fat and a reduction in lean body mass at 3 weeks of age (Figure 6, A). At 24 weeks, all 3 groups exposed to an HF diet during pregnancy and/or lactation demonstrated markedly increased percentage body fat as compared with Con/Con (Figure 6, B). However, as described previously, only the HF/HF and Con/HF demonstrated significantly increased body mass. The HF/Con males and females had significantly reduced lean body mass as compared with respective Con/Con offspring, as measured by both grams and percentage body weight, whereas all 3 groups exposed to an HF diet during pregnancy and/or lactation demonstrated a reduced percentage lean body mass at 24 weeks.
      Figure thumbnail gr6
      Figure 6Body composition of 3 and 24 week old offspring
      A, Body mass, percentage body fat, and percentage lean body mass in 3 week old offspring. Because no sex differences were evident, combined data of males (n = 8) and females (n = 8) from 8 litters in the 4 groups are shown. Values are mean ± SE. B, Body mass, lean body mass, and percentage body fat and lean body mass in 24 week old male (light gray columns) and female (white columns) offspring from 4 groups. Number of offspring studied per group was 8 males and 8 females from 8 litters. Values are mean ± SE.
      Con, control; HF, high-fat.
      P < .001 vs Con offspring; $P < .001 males vs females.
      Desai. Maternal obesity programs offspring metabolic syndrome. Am J Obstet Gynecol 2014.

      Plasma profile

      At 1 day of age, HF newborns had lower plasma triglyceride, cholesterol, leptin, and corticosterone levels, although similar plasma insulin levels as Con/Con (Table 4).
      Table 4Plasma profile of newborn pups
      VariableConHF
      Body weight, g7.3 ± 0.17.4 ± 0.2
      Plasma triglyceride, mg/dL126 ± 992 ± 12
      P < .05 vs Con.
      Plasma cholesterol, mg/d L95.7 ± 2.782.5 ± 3.3
      P < .05 vs Con.
      Plasma leptin, ng/mL4.9 ± 0.92.1 ± 0.5
      P < .05 vs Con.
      Plasma insulin, ng/mL0.52 ± 0.100.52 ± 0.20
      Plasma corticosterone, ng/mL80 ± 1252 ± 8
      P < .05 vs Con.
      Plasma profile of one day old Con and HF pups. Because no sex differences were evident, combined data of males (n = 16) and females (n = 16) from 16 litters in the 4 groups are shown.
      Con, control; HF, high-fat.
      Desai. Maternal obesity programs offspring metabolic syndrome. Am J Obstet Gynecol 2014.
      a P < .05 vs Con.
      At 3 weeks of age, plasma triglyceride levels were significantly increased among HF/HF and Con/HF offspring, although there was no difference in plasma cholesterol levels. However, HF/HF and HF/Con offspring both demonstrated significantly increased plasma leptin, insulin, and corticosterone levels as compared with Con/Con and Con/HF. Consistent with the plasma insulin levels, HF/HF and HF/Con offspring demonstrated elevated blood glucose levels (Figure 7, A).
      Figure thumbnail gr7a
      Figure 7Plasma profile of 3 and 24-week-old offspring
      A, Plasma triglyceride, cholesterol, glucose, leptin, corticosterone, and insulin levels in 3 week old offspring. Because no sex differences were evident, combined data of males (n = 8) and females (n = 8) from 8 litters in the 4 groups are shown. Values are mean ± SE. B, Plasma triglyceride, cholesterol, glucose, leptin, corticosterone, and insulin levels in 24-week-old male (light gray columns) and female (white columns) offspring from 4 groups. Number of offspring studied per group was 8 males and 8 females from 8 litters. Values are mean ± SE.
      Con, control.
      P < .01 vs Con offspring; Asterisk indicates P < .001 vs Con offspring. $P < .001 males vs females.
      Desai. Maternal obesity programs offspring metabolic syndrome. Am J Obstet Gynecol 2014.
      Figure thumbnail gr7b
      Figure 7Plasma profile of 3 and 24-week-old offspring
      A, Plasma triglyceride, cholesterol, glucose, leptin, corticosterone, and insulin levels in 3 week old offspring. Because no sex differences were evident, combined data of males (n = 8) and females (n = 8) from 8 litters in the 4 groups are shown. Values are mean ± SE. B, Plasma triglyceride, cholesterol, glucose, leptin, corticosterone, and insulin levels in 24-week-old male (light gray columns) and female (white columns) offspring from 4 groups. Number of offspring studied per group was 8 males and 8 females from 8 litters. Values are mean ± SE.
      Con, control.
      P < .01 vs Con offspring; Asterisk indicates P < .001 vs Con offspring. $P < .001 males vs females.
      Desai. Maternal obesity programs offspring metabolic syndrome. Am J Obstet Gynecol 2014.
      At 24 weeks of age, plasma triglyceride levels were increased among the 3 groups of HF exposure, although only Con/HF males demonstrated increased cholesterol levels. Furthermore, all 3 groups of HF exposure showed increased levels of glucose, leptin, and insulin levels in both sexes. However, plasma coritcosterone levels were significantly elevated in HF/HF and HF/Con male and female offspring (Figure 7, B).

      Glucose tolerance test

      At 6 weeks of age, GTTs were performed in all offspring. HF/Con males and females demonstrated the greatest area under the curve as compared with the other 3 groups (Figure 8, A). When the GTT was repeated at 24 weeks of age, HF/Con offspring again demonstrated the greatest areas under the curve, with intermediate values evident among HF/HF and Con/HF as compared with Con/Con.
      Figure thumbnail gr8
      Figure 8Glucose tolerance test at 6 and 24 weeks of age
      A, Mean glucose values of male and female offspring at 6 weeks of age from Con (filled circles; pups born to Con dams and nursed by Con dams), HF/HF (open circles; pups born to HF dams and nursed by HF dams), HF/Con (open triangles; pups born to HF dams and nursed by Con dams), and Con/HF (filled triangles; pups born to Con dams and nursed by HF dams). The GTT area under the curve of male (closed squares) and female (open squares) offspring from 4 groups is shown. Number of offspring studied per group was 8 males and 8 females (from 8 litters). Values are mean ± SE. B, Mean glucose values of male and female offspring at 24 weeks of age from Con (filled circles; pups born to Con dams and nursed by Con dams), HF/HF (open circles; pups born to HF dams and nursed by HF dams), HF/Con (open triangles; pups born to HF dams and nursed by Con dams), and Con/HF (filled triangles; pups born to Con dams and nursed by HF dams). The GTT area under the curve of male (closed squares) and female (open squares) offspring from four groups is shown. Number of offspring studied per group was 8 males and 8 females (from 8 litters). Values are mean ± SE.
      Con, control; GTT, glucose tolerance test; HF, high-fat.
      P < .05.
      Desai. Maternal obesity programs offspring metabolic syndrome. Am J Obstet Gynecol 2014.

      Blood pressure

      Among 8 week old male and female offspring, elevated systolic blood pressures were demonstrated in males and females of all 3 groups exposed to the HF diet during pregnancy and/or lactation (HF/HF, HF/Con, and Con/HF) as compared with Con/Con (Figure 9). We were unable to measure blood pressure at 24 weeks of age because of the unavailability of tail-cuff size in that range.
      Figure thumbnail gr9
      Figure 9Systolic blood pressure
      Systolic blood pressure in 8 week old male (closed squares) and female (open squares) offspring from 4 groups. Number of offspring studied per group was 8 males and 8 females from 8 litters. Values are mean ± SE.
      Con, control; HF, high-fat.
      P < .001 vs Con offspring.
      Desai. Maternal obesity programs offspring metabolic syndrome. Am J Obstet Gynecol 2014.

      Male/female comparison

      In general, 24 week old male offspring had significantly higher percentage lean body mass, blood pressure, plasma leptin, insulin, and triglyceride levels as compared with female offspring. In contrast, percentage body fat and plasma corticosterone levels were significantly higher in females than males.

      Comment

      The results of the present study demonstrate a marked impact of maternal obesity/HF diet on offspring body composition and the risk of metabolic syndrome. Importantly, differential effects on offspring phenotype were observed dependent on whether exposure to maternal obesity occurred during pregnancy, during lactation, or both the pregnancy and lactation periods.
      Although HF dams were significantly heavier at conception and throughout pregnancy, both HF and Con dams gained similar amounts of weight during pregnancy and demonstrated similar total caloric intakes. Recent guidelines for gestational weight gain have advocated lower weight gain among overweight or obese patients as opposed to normal weight women,
      • Thangaratinam S.
      • Rogozińska E.
      • Jolly K.
      • et al.
      Interventions to reduce or prevent obesity in pregnant women: a systematic review.
      • Rasmussen K.M.
      • Abrams B.
      • Bodnar L.M.
      • Butte N.F.
      • Catalano P.M.
      • Maria Siega-Riz A.
      Recommendations for weight gain during pregnancy in the context of the obesity epidemic.
      with some studies advocating a zero weight gain goal among morbidly obese patients.
      • Vesco K.K.
      • Karanja N.
      • King J.C.
      • et al.
      Healthy Moms, a randomized trial to promote and evaluate weight maintenance among obese pregnant women: study design and rationale.
      Extrapolation from the present rodent studies to humans would suggest that despite an approximately 33% increment in prepregnancy body weight, there is an intrinsic programmed increment in pregnancy weight gain because, in part, of fetal mass, maternal fat deposition, and maternal physiological alterations (eg, blood volume expansion). Thus, attempts to limit maternal body weight gain may require active intervention strategies.
      Notably, Con dams lost more weight at delivery than HF dams, although the explanation for this is unclear. Because litter size and pup weight were similar between the 2 groups, this weight loss is not a reflection of fetal mass. Alternatively, changes in maternal physiological adaptations (eg, maternal blood volume alterations) or in the amniotic/allantois fluid compartments may contribute to the increased maternal weight loss in Con dams.
      Con dams nursing either Con or HF pups demonstrated a gradual increase in body weight through the first half of lactation and a decrease in body weight during the last third of lactation, likely associated with the increased pup milk ingestion and pup weight gain. Similar findings were observed in the HF dams nursing HF pups. Surprisingly, the HF dams nursing Con pups demonstrated continued weight loss throughout the lactation period. Nevertheless, both HF dams nursing Con or HF pups demonstrated a marked increase in caloric intake in the last third of lactation.
      This caloric intake likely served to support the increased weight gain demonstrated by both HF/HF pups and the Con/HF pups. As clarified in the Material and Methods section, because of the process we utilized for identifying dams and pups, Con/HF pups were born to Con dams and nursed by HF dams. These findings indicate that maternal caloric intake, and thus, maintenance of maternal body weight is dependent, at least in part, on the offspring weight gain during lactation.
      The mechanism for the loss of maternal body weight in the HF dams nursing Con pups is unclear because the weight gain of these pups was identical with that of the HF/HF pups, and the caloric intakes of the dams were also identical. Nevertheless, the increased weight loss of HF dams nursing Con pups during lactation was reflected in their body composition and plasma profile because these dams demonstrated a significantly reduced percentage body fat, plasma leptin, and plasma insulin as compared with the HF dams nursing HF pups at day 21 of lactation.
      Consistent with human pregnancy, both Con and HF dams demonstrated elevated plasma cholesterol and triglyceride levels as compared with nonpregnant animals
      • Salameh W.A.
      • Mastrogiannis D.S.
      Maternal hyperlipidemia in pregnancy.
      • Herrera E.
      Lipid metabolism in pregnancy and its consequences in the fetus and newborn.
      • Ramsay J.E.
      • Ferrell W.R.
      • Crawford L.
      • Wallace A.M.
      • Greer I.A.
      • Sattar N.
      Maternal obesity is associated with dysregulation of metabolic, vascular, and inflammatory pathways.
      and compared with end-of-lactation values. Despite the HF diet throughout pregnancy and lactation, both HF dams nursing HF and Con pups demonstrated lower plasma triglyceride and cholesterol levels as compared with Con dams nursing Con or HF pups. Thus, the HF dams transitioned from significantly elevated plasma cholesterol and triglycerides at day 20 of pregnancy to significantly lower values of both analytes at the completion of lactation, despite identical diets during both periods. Thus, it is not dietary changes, but rather metabolism and perhaps gastrointestinal absorption, that contribute to these basal levels of cholesterol and triglycerides.
      It is known that lipid metabolism differs between lean and obese pregnant women, with obese women demonstrating an earlier shift from an anabolic to a catabolic state and a predominance of lipolysis as compared with lean women.
      • Heerwagen M.J.
      • Miller M.R.
      • Barbour L.A.
      • Friedman J.E.
      Maternal obesity and fetal metabolic programming: a fertile epigenetic soil.
      • Catalano P.M.
      • Roman-Drago N.M.
      • Amini S.B.
      • Sims E.A.
      Longitudinal changes in body composition and energy balance in lean women with normal and abnormal glucose tolerance during pregnancy.
      Although maternal hypercholesterolemia and hypertriglyceridemia during pregnancy is believed to contribute to the long-term risk of atherosclerosis or hypertension,
      • Palinski W.
      • Yamashita T.
      • Freigang S.
      • Napoli C.
      Developmental programming: maternal hypercholesterolemia and immunity influence susceptibility to atherosclerosis.
      • Napoli C.
      • D'Armiento F.P.
      • Mancini F.P.
      • et al.
      Fatty streak formation occurs in human fetal aortas and is greatly enhanced by maternal hypercholesterolemia. Intimal accumulation of low density lipoprotein and its oxidation precede monocyte recruitment into early atherosclerotic lesions.
      elevated cholesterol and fatty acid profiles during what may be years of lactation may also have an associated risk. Thus, the reduced levels of triglycerides and cholesterol in overweight and obese HF dams during lactation deserves further study to explore the mechanisms regulating cholesterol and fatty acid profiles.
      Notably, plasma corticosterone levels were markedly elevated in HF dams at term, indicating that the effects of diet and/or maternal obesity represented a significant stress factor. HF dams continued to have elevated plasma corticosterone levels at day 21 of lactation, regardless of the pup identification. However, the highest levels of plasma corticosterone were observed in Con dams nursing HF pups, perhaps suggesting that the increased weight gain of these pups (HF/Con) induces a significant maternal stress. Previous studies on rat maternal obesity have made a similar observation of elevated maternal and lower offspring coritcosterone levels.
      • Rodriguez J.S.
      • Rodríguez-González G.L.
      • Reyes-Castro L.A.
      • et al.
      Maternal obesity in the rat programs male offspring exploratory, learning and motivation behavior: prevention by dietary intervention pre-gestation or in gestation.
      Examination of offspring weight gain demonstrates programming effects of maternal diet during both pregnancy and lactation. Consumption of an HF diet during pregnancy did not have an impact on birthweight and, if limited to the pregnancy, did not have an impact on the body weights of either males or females as 24 week adults. Consumption of an HF diet during lactation only was associated with increased body weight of male and female offspring at 24 weeks, suggesting that an HF diet does not impair milk production. However, consumption of a HF diet during both pregnancy and lactation (HF/HF) resulted in a further increase of body weight of males, although not females, as compared with respective Con/HF at 24 weeks. Thus, the pregnancy HF diet potentiates offspring growth when offspring are also exposed to dams consuming a HF diet during nursing.
      Offspring food intake appears to be a major contributor to body weight changes in males because food intake paralleled changes in body weight. Although female food intake showed a similar trend, the lack of statistical difference in food intake despite significant differences in body weight to 24 weeks may suggest that either metabolic efficiency and/or energy expenditure may be programmed by maternal dietary changes during pregnancy and/or lactation. Notably, studies on rodents and sheep offspring suggested a programming effect of maternal diet on energy expenditure in adult offspring.
      • Vickers M.H.
      • Breier B.H.
      • McCarthy D.
      • Gluckman P.D.
      Sedentary behavior during postnatal life is determined by the prenatal environment and exacerbated by postnatal hypercaloric nutrition.
      • Gardner D.S.
      • Rhodes P.
      Developmental origins of obesity: programming of food intake or physical activity?.
      • Donovan E.L.
      • Hernandez C.E.
      • Matthews L.R.
      • et al.
      Periconceptional undernutrition in sheep leads to decreased locomotor activity in a natural environment.
      The increased body weight of HF/HF and Con/HF offspring at 24 weeks was associated with increased percentage body fat of both males and females. This was accompanied by reduced absolute and percentage lean body mass in Con/HF males and females. These findings are similar to that described in humans, particularly in the Indian subcontinent, of the thin-fat phenotype in which individuals have a normal body weight but markedly increased body fat and reduced lean body mass.
      • Yajnik C.S.
      Early life origins of insulin resistance and type 2 diabetes in India and other Asian countries.
      Importantly, it was subscapular rather than abdominal visceral fat that was preserved, suggesting increased subcutaneous adipose tissue.
      Similarly, a study by Zambrano et al
      • Zambrano E.
      • Martínez-Samayoa P.M.
      • Rodríguez-González G.L.
      • Nathanielsz P.W.
      Dietary intervention prior to pregnancy reverses metabolic programming in male offspring of obese rats.
      (2010) showed increased subcutaneous fat in offspring exposed to maternal obesity during the fetal and nursing periods. However in the present study, it remains to be determined whether there is a differential effect on visceral vs subcutaneous adipose tissue as a result of an HF diet exposure during pregnancy and/or lactation.
      • Zambrano E.
      • Martínez-Samayoa P.M.
      • Rodríguez-González G.L.
      • Nathanielsz P.W.
      Dietary intervention prior to pregnancy reverses metabolic programming in male offspring of obese rats.
      Notably, all 3 groups exposed to an HF diet during any portion of pregnancy or lactation demonstrated a significant decrease in percentage lean body mass as compared with Con offspring.
      Despite their normal birthweight, HF pups had lower plasma triglyceride, cholesterol, leptin, and coritcosterone levels. Moreover, these values contrasted with elevated maternal levels. Although the lower HF newborn plasma levels are consistent with previous studies,
      • Howie G.J.
      • Sloboda D.M.
      • Kamal T.
      • Vickers M.H.
      Maternal nutritional history predicts obesity in adult offspring independent of postnatal diet.
      • Morris M.J.
      • Chen H.
      Established maternal obesity in the rat reprograms hypothalamic appetite regulators and leptin signaling at birth.
      the mechanism remains unclear and may involve placental effects. Unlike the newborn, the plasma profile of 3 week old offspring reflected the maternal diet during lactation with HF/HF and Con/HF offspring demonstrating elevated triglyceride levels and increased leptin. However, at 24 weeks, all groups exposed to HF during pregnancy, lactation, or both demonstrated elevated plasma triglyceride and plasma leptin levels. Thus, HF during pregnancy alone results in adult, although not weanling, hypertriglyceridemia.
      Plasma corticosterone levels were elevated in 3 week old HF/HF and HF/Con offspring, and these effects persisted at 24 weeks, again indicating the programming effects of the HF diet during pregnancy on offspring endogenous glucocorticoid expression. Similarly, the pregnancy HF diet resulted in elevated blood glucose and plasma insulin levels at 3 weeks of age (HF/HF HF/Con), although all 3 HF-exposed groups demonstrated elevated blood glucose and plasma insulin at 24 weeks. Despite the elevated glucose and insulin levels in all 3 groups of HF-exposed offspring, only the HF/Con offspring demonstrated an elevated area under the glucose tolerance test at 6 weeks of age. Nevertheless, these results indicate that the augmented insulin secretion in HF/Con is not adequate for the maintenance of normoglycemia in this group.
      Consistent with elevated blood glucose and plasma insulin, impaired glucose tolerance with elevated area under the curve in all 3 HF offspring groups is evident at 24 weeks of age. The markedly elevated plasma insulin levels in HF/HF offspring at 24 weeks is nearly 2-fold that of HF/Con and Con/HF. These results suggest that the latter 2 groups may have inadequate insulin secretion contributing to the plasma hyperglycemia evident at 24 weeks, whereas the HF/HF offspring have marked hyperglycemia and hyperinsulinemia, indicative of insulin-resistant diabetes. Similar characteristics of increased adiposity and insulin resistance have been reported in adult offspring exposed to maternal HF diet prior to mating and throughout pregnancy and lactation.
      • Zambrano E.
      • Martínez-Samayoa P.M.
      • Rodríguez-González G.L.
      • Nathanielsz P.W.
      Dietary intervention prior to pregnancy reverses metabolic programming in male offspring of obese rats.
      All groups of male and female HF-exposed offspring demonstrated elevated systolic pressure at 8 weeks of age. As noted previously, plasma corticosterone levels were elevated in HF/HF and HF/Con offspring, suggesting that elevated glucocorticoids may contribute to the mechanism of hypertension in this group.
      • O'Regan D.
      • Kenyon C.J.
      • Seckl J.R.
      • Holmes M.C.
      Glucocorticoid exposure in late gestation in the rat permanently programs gender-specific differences in adult cardiovascular and metabolic physiology.
      However, the mechanism of hypertension in Con/HF offspring likely involves mechanisms other than glucocorticoid-mediated processes. Because vasculogenesis continues during the nursing period of maternal HF exposure, it is possible that alterations in vascular and/or cardiac development may alter blood pressure regulation.
      • Tokunaga H.
      Postnatal development of the blood vasculature in the rat adrenal gland: a scanning electron microscope study of microcorrosion casts.
      • Armitage J.A.
      • Lakasing L.
      • Taylor P.D.
      • et al.
      Developmental programming of aortic and renal structure in offspring of rats fed fat-rich diets in pregnancy.
      • Dong M.
      • Zheng Q.
      • Ford S.P.
      • Nathanielsz P.W.
      • Ren J.
      Maternal obesity, lipotoxicity and cardiovascular diseases in offspring.
      The overall sex differences, namely that the males exhibit higher lean body mass, blood pressure, and plasma triglyceride levels than females, is consistent with prior human and rat data.
      • Bayol S.A.
      • Simbi B.H.
      • Bertrand J.A.
      • Stickland N.C.
      Offspring from mothers fed a “junk food” diet in pregnancy and lactation exhibit exacerbated adiposity that is more pronounced in females.
      • Freedman D.S.
      • Jacobsen S.J.
      • Barboriak J.J.
      • et al.
      Body fat distribution and male/female differences in lipids and lipoproteins.
      The disparity in sex response to the maternal obesity/HF diet was evident only in the offspring exposed exclusively during the nursing period (Con/HF). This was apparent only in the cholesterol levels, with the males exhibiting higher levels. Although the cause for sex-dependent response is unknown, previous studies have noted a differential impact of early overnutrition on males vs females.
      • Elahi M.M.
      • Cagampang F.R.
      • Mukhtar D.
      • Anthony F.W.
      • Ohri S.K.
      • Hanson M.A.
      Long-term maternal high-fat feeding from weaning through pregnancy and lactation predisposes offspring to hypertension, raised plasma lipids and fatty liver in mice.
      The primary mechanism linking fetal/postnatal growth with glucose impairment and dyslipidemia remains to be determined, although studies have implicated alterations in epigenomics, gene expression, and signaling factors, including alterations in the milk composition.
      • Murabayashi N.
      • Sugiyama T.
      • Zhang L.
      • et al.
      Maternal high-fat diets cause insulin resistance through inflammatory changes in fetal adipose tissue.
      • Aagaard-Tillery K.M.
      • Grove K.
      • Bishop J.
      • et al.
      Developmental origins of disease and determinants of chromatin structure: maternal diet modifies the primate fetal epigenome.
      • Purcell R.H.
      • Sun B.
      • Pass L.L.
      • Power M.L.
      • Moran T.H.
      • Tamashiro K.L.
      Maternal stress and high-fat diet effect on maternal behavior, milk composition, and pup ingestive behavior.
      The mechanism of programming resulting from in utero overnutrition likely involves an interacting effect of preexisting maternal obesity and HF diet effects.
      In summary, the maternal phenotype during pregnancy and the end of lactation evidenced markedly elevated body fat and plasma corticosterone levels in HF dams. The results of the present study demonstrate that the maternal HF diet during lactation promotes offspring obesity, and the HF diet during pregnancy and lactation results in a further increase in offspring body weight and percentage fat mass. The HF diet during pregnancy alone programmed increased offspring adiposity (percentage body fat), although with normal body weight, simulating the thin/fat human phenotype.
      Metabolic disturbances, particularly that of hyperglycemia, were apparent in all groups exposed to a maternal HF diet (during pregnancy and/or lactation), although differences were apparent in the manifestation of insulin-resistant vs insulin-deficient phenotypes. Elevated systolic blood pressure was manifest in all groups, implying that exposure to an obese/HF environment is disadvantageous for offspring health, regardless of pregnancy or lactation periods. Nonetheless, the underlying mechanism may differ because offspring that experienced in utero HF exposure had increased corticosterone levels. Thus, for both fetal and maternal well-being, prepregnancy obesity and a Western HF diet may have a significant adverse impact on long-term health. The exposure to an HF diet during both pregnancy and lactation results in an altered phenotype and exaggerated obesity when compared with pregnancy alone. However, pregnancy exposure alone also has significant adverse effects, indicating that suggestions to limit nursing or advocate formula feeding to infants of obese/HF mothers would likely not remedy the programming effects of the pregnancy environment. Rather, preconceptual weight loss is optimal to promote the beneficial effects of a pregnancy environment.

      Acknowledgments

      We thank Stacy Behare and Linda Day for technical assistance.

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