KLF4 down-regulation underlies placental angiogenesis impairment induced by maternal glucose intolerance in late pregnancy.

Maternal glucose intolerance in late pregnancy can easily impair pregnancy outcomes and placental development. The impairment of placental angiogenesis is closely related to the occurrence of glucose intolerance during pregnancy, but the mechanism remains largely unknown. In this study, the pregnant mouse model of maternal high-fat diet and endothelial injury model of porcine vascular endothelial cells (PVECs) was used to investigate the effect of glucose intolerance on pregnancy outcomes and placental development. Feeding pregnant mice, a high-fat diet was shown to induce glucose intolerance in late pregnancy, and significantly increase the incidence of resorbed fetuses. Moreover, a decrease was observed in the proportion of blood sinusoids area and the expression level of CD31 in placenta, indicating that placental vascular development was impaired by high-fat diet. Considering that hyperglycemia is an important symptom of glucose intolerance, we exposed PVECs to high glucose (50 mM), which verified the negative effects of high glucose on endothelial function. Bioinformatics analysis further emphasized that high glucose exposure could significantly affect the angiogenesis-related functions of PVECs and predicted that Krüppel-like factor 4 (KLF4) may be a key mediator of these functional changes. The subsequent regulation of KLF4 expression confirmed that the inhibition of KLF4 expression was an important reason why high glucose impaired the endothelial function and angiogenesis of PVECs. These results indicate that high-fat diet can aggravate maternal glucose intolerance and damage pregnancy outcome and placental angiogenesis, and that regulating the expression of KLF4 may be a potential therapeutic strategy for maintaining normal placental angiogenesis.

Starch supplementation improves the reproductive performance of sows in different glucose tolerance status.

This study was to evaluate the effects of glucose tolerance status, maternal starch supplementation and soybean substitution in diets on the performance of dams and their offspring. Eighty-eight pregnant sows (Landrace × Large White) were selected from an initial total of 120 sows, based on blood glucose test values, and assigned to 4 experimental treatments in a 2 × 2 factorial design. The factors were glucose tolerance status (glucose intolerant [GIT] vs. normal glucose tolerant [NGT]) or dietary treatments (corn starch diet [CS] vs. soybean substitution diet [SS]). A higher area under the curve (AUC) for post-meal glucose was observed (P < 0.05) in the GIT group than in the NGT group on d 109 of gestation. The CS group had a lower value of homeostasis model assessment-insulin resistance than the SS group (P < 0.05) on d 109 of gestation. Corn starch supplementation for sows decreased the stillbirth rate (P < 0.05), regardless of the sows' glucose tolerance status. The villus height of the jejunum and the villus height to crypt depth ratio of the ileum were greater in normal birth weight piglets from the CS group than from the SS group (P < 0.01), and so was the activity of sucrase in the jejunum and ileum (P < 0.01). Compared with the SS group, the CS group showed a reduction in pre-weaning mortality rate, an increase in the number of high-birth-weight piglets, and a decrease in the number of low-birth-weight piglets (P < 0.05) under GIT status. In conclusion, sows fed CS decreased stillbirth rate and improved insulin resistance, as well as improving the intestinal morphology and digestive enzyme activities of their progeny, regardless of glucose tolerance status. Additionally, the CS group improved birth weight distribution and decreased pre-weaning mortality rate of piglets under GIT status.

A maternal high-fat/low-fiber diet impairs glucose tolerance and induces the formation of glycolytic muscle fibers in neonatal offspring.

PURPOSE: In our previous study, the maternal high-fat/low-fiber (HF-LF) diet was suggested to induce metabolic disorders and placental dysfunction of the dam, but the effects of this diet on glucose metabolism of neonatal offspring remain largely unknown. Here, a neonatal pig model was used to evaluate the effects of maternal HF-LF diet during pregnancy on glucose tolerance, transition of skeletal muscle fiber types, and mitochondrial function in offspring. METHODS: A total of 66 pregnant gilts (Guangdong Small-ear Spotted pig) at day 60 of gestation were randomly divided into two groups: control group (CON group; 2.86% crude fat, 9.37% crude fiber), and high-fat/low-fiber diet group (HF-LF group; 5.99% crude fat, 4.13% crude fiber). RESULTS: The maternal HF-LF diet was shown to impair the glucose tolerance of neonatal offspring, downregulate the protein level of slow-twitch fiber myosin heavy chain I (MyHC I), and upregulate the protein levels of fast-twitch fiber myosin heavy chain IIb (MyHC IIb) and IIx (MyHC IIx) in soleus muscle. Additionally, compared with the CON group, the HF-LF offspring showed inhibition of insulin signaling pathway and decrease in mitochondrial function in liver and soleus muscle. CONCLUSION: Maternal HF-LF diet during pregnancy impairs glucose tolerance, induces the formation of glycolytic muscle fibers, and decreases the hepatic and muscular mitochondrial function in neonatal piglets.

Placentae for Low Birth Weight Piglets Are Vulnerable to Oxidative Stress, Mitochondrial Dysfunction, and Impaired Angiogenesis.

Intrauterine growth restriction (IUGR) is associated with fetal mortality and morbidity. One of the most common causes of IUGR is placental insufficiency, including placental vascular defects, and mitochondrial dysfunction. In addition, a high level of oxidative stress induces placental vascular lesions. Here, we evaluated the oxidative stress status, mitochondrial function, angiogenesis, and nutrient transporters in placentae of piglets with different birth weights: <500 g (L), 500-600 g (LM), 600-700 g (M), and >700 g (H). Results showed that placentae from the L group had higher oxidative damage, lower adenosine triphosphate and citrate synthase levels, and lower vascular density, compared to those from the other groups. Protein expression of angiogenic markers, including vascular endothelial cadherin, vascular endothelial growth factor A, and platelet endothelial cell adhesion molecule-1, was the lowest in the L group placentae compared to the other groups. In addition, the protein levels of glucose transporters GLUT1 and GLUT3 were downregulated in the L group, compared to the other groups. Furthermore, oxidative stress induced by H2O2 inhibited tube formation and migration in porcine vascular endothelial cells. Collectively, placentae for lower birth weight neonates are vulnerable to oxidative damage, mitochondrial dysfunction, and impaired angiogenesis.

Effects of different maternal feeding strategies from day 1 to day 85 of gestation on glucose tolerance and muscle development in both low and normal birth weight piglets.

BACKGROUND: Maternal nutrition during gestation plays a vital role in fetal development. The effects of different maternal feeding strategies from day 1 to day 85 of gestation on glucose tolerance and muscle development in low and normal birth weight offspring were investigated by using 80 gilts randomly allotted to T1 and T2 groups and treated respectively with a gradual-increase (T1) and a convex transition (T2) feeding strategy, with no difference in total feed intake. RESULTS: T2 group was seen to have a higher percentage of piglets with birth weight less than 500 g, while T1 group was shown to have a higher percentage of piglets with birth weight over 700 g. Meanwhile, for both low and normal birth weight piglets, T1 group was higher than T2 group in terms of muscle free amino acid concentration, mRNA expression levels of muscle growth-related factors, relative muscle fiber number and cross-sectional area. We must emphasize that the T2 group was shown to improve glucose tolerance, slow-twitch muscle fiber protein levels, and muscle mitochondrial function only in low birth weight piglets. CONCLUSION: The convex transition feeding strategy can decrease the percentage of piglets with birth weight over 700 g, while improving glucose tolerance, slow-twitch muscle fiber protein levels, and muscle mitochondrial function in low birth weight piglets. Our findings provide new evidence for the potential importance of nutritional strategies during gestation, especially for improving the glucose tolerance and muscle development of low birth weight neonatal. © 2020 Society of Chemical Industry.

Maternal Diet-Induced Obesity Compromises Oxidative Stress Status and Angiogenesis in the Porcine Placenta by Upregulating Nox2 Expression.

Maternal obesity is associated with placental oxidative stress. However, the mechanism underlying this association remains poorly understood. In the present study, a gilt obesity model was developed by exposure to different energy diets and used to investigate the role of NADPH oxidase 2 (Nox2) in the placenta. Specifically, 99 gilts (Guangdong Small-ear Spotted pig) at day 60 of gestation were randomly assigned to one of the following three treatments: low-energy group (L, DE = 11.50 MJ/kg), medium-energy group (M, DE = 12.41 MJ/kg), and high-energy group (H, DE = 13.42 MJ/kg), with 11 replicate pens per treatment and 3 gilts per pen. At the start of the study, maternal body weight and backfat thickness were not significantly different in the three treatments. After the study, data indicated that the H group had higher body weight and backfat thickness gain for gilts during gestation and lower piglet birth weight compared with the other two groups. Additionally, the H group showed glucolipid metabolic disorders and increased triglyceride and nonesterified fatty acid contents in the placenta of gilts. Compared with the L group, the H group exhibited lower mitochondrial biogenesis and increased oxidative damage in the placenta. Importantly, increased mRNA expression and protein abundance of Nox2 were observed for the first time in H group placentae. Furthermore, compared with the L group, the H group showed a decrease in the density of placental vessels and the protein levels of vascular endothelial cadherin (VE-cadherin), vascular endothelial growth factor A (VEGF-A), and phosphorylation of vascular endothelial growth factor receptor 2 (p-VEGFR2) as well as the immunostaining intensity of platelet endothelial cell adhesion molecule-1 (CD31). Our findings suggest that maternal high-energy diet-induced obesity increases placental oxidative stress and decreases placental angiogenesis possibly through the upregulation of Nox2.

Dietary energy sources during late gestation and lactation of sows: effects on performance, glucolipid metabolism, oxidative status of sows, and their offspring1.

In this study, the effects of maternal energy sources during late gestation and lactation on the performance, glucolipid metabolism, and oxidative status of sows and their offspring were investigated using a total of 75 (2 to 6 of parity) Landrace × Large White sows at day 85 of gestation under 3 different dietary treatments: SO diet (basal diet plus 3.0% and 5.0% soybean oil during late gestation and lactation, respectively), FO diet (basal diet plus 3.0%/5.0% fish oil during late gestation and lactation, respectively), and CS diet (basal diet plus 32%/42% corn starch during late gestation and lactation, respectively). All the 3 groups showed no obvious differences (P > 0.05) in the number of total piglets born, born alive, after cross-fostering, and at weaning, whereas the CS group exhibited a shorter farrowing duration (P < 0.05) and lower stillbirth rate (P < 0.05) when compared with the SO group. In addition, litter weight at birth was significantly higher in the CS group than in the SO or FO group (P < 0.05). Despite no notable differences in the ADG of suckling piglets among dietary treatments (P > 0.05), the CS group had greater feed intake than the SO group during the lactation period (P < 0.05). In neonatal piglets with normal birth weight (NBW, 1.3 to 1.5 kg), the CS group was lower than the SO group in the content of liver glycogen (P < 0.05) and the mRNA abundances of fatty acid synthase, acetyl-CoA carboxylase, fatty acid-binding protein 1, and acyl-CoA oxidase (P < 0.05). Interestingly, compared with the SO group, the FO group had a lower preweaning mortality rate (P < 0.05), but greater liver glycogen pools (P < 0.05) in neonatal piglets with low birth weight (LBW, <1.1 kg). Compared with the CS group, the FO group showed an increase in the plasma malondialdehyde levels (P < 0.05) of sows, as well as an increase of 8-hydroxy-deoxyguanosine (P < 0.05) and a decrease of ferric reducing ability of plasma (P < 0.05) in NBW piglets. Overall, the diet rich in starch decreased the stillbirth rate and increased the litter weight of neonatal piglets, the dietary supplementation with fish oil decreased preweaning mortality rate, and the diet with a low n6:n3 ratio increased the oxidative status of sows and their offspring.