Dietary bile acid supplementation in weaned piglets with intrauterine growth retardation improves colonic microbiota,metabolic activity,and epithelial function

Background Intrauterine growth retardation(IUGR)is one of the major constraints in animal production.Our previ-ous study showed that piglets with IUGR are associated with abnormal bile acid(BA)metabolism.This study explored whether dietary BA supplementation could improve growth performance and colonic development,function,micro-biota,and metabolites in the normal birth weight(NBW)and IUGR piglets.A total of 48 weaned piglets(24 IUGR and 24 NBW)were allocated to four groups(12 piglets per group):(i)NBW group,(ii)NBW+BA group,(iii)IUGR group,and(iv)IUGR+BA group.Samples were collected after 28 days of feeding.Results The results showed that dietary BA supplementation increased the length and weight of the colon and colon weight to body weight ratio,while decreased the plasma diamine oxidase(DAO)concentration in the NBW pig-lets(P<0.05).Dietary BA supplementation to IUGR piglets decreased(P<0.05)the plasma concentrations of D-lactate and endotoxin and colonic DAO and endotoxin,suggesting a beneficial effect on epithelial integrity.Moreover,dietary BA supplementation to NBW and IUGR piglets increased Firmicutes abundance and decreased Bacteroidetes abundance(P<0.05),whereas Lactobacillus was the dominant genus in the colon.Metabolome analysis revealed 65 and 51 differential metabolites in the colon of piglets fed a diet with/without BA,respectively,which was associated with the colonic function of IUGR piglets.Furthermore,dietary BA supplementation to IUGR piglets upregulated the expressions of CAT,GPX,SOD,Nrf1,IL-2,and IFN-γin colonic mucosa(P<0.05).Conclusions Collectively,dietary BA supplementation could improve the colonic function of IUGR piglets,which was associated with increasing proportions of potentially beneficial bacteria and metabolites.Furthermore,BA shows a promising application prospect in improving the intestinal ecosystem and health of animals.

Pterostilbene attenuates intrauterine growth retardation-induced colon inflamm tion in piglets by modulating endoplasmic reticulum stress and autophagy

Background:Endoplasmic reticulum(ER)stress and autophagy are implicated in the pathophysiology of intestinal inflammation;however,their roles in intrauterine growth retardation(IUGR)-induced colon inflammation are unclear.This study explored the protective effects of natural stilbene pterostilbene on colon inflammation using the IUGR piglets and the tumor necrosis factor alpha(TNF-α)-treated human colonic epithelial cells(Caco-2)by targeting ER stress and autophagy.Results:Both the IUGR colon and the TNF-α-treated Caco-2 cells exhibited inflammatory responses,ER stress,and impaired autophagic flux(P<0.05).The ER stress inducer tunicamycin and the autophagy inhibitor 3-methyladenine further augmented inflammatory responses and apoptosis in the TNF-α-treated Caco-2 cells(P<0.05).Conversely,pterostilbene inhibited ER stress and restored autophagic flux in the IUGR colon and the TNF-α-treated cells(P<0.05).Pterostilbene also prevented the release of inflammatory cytokines and nuclear translocation of nuclear factor kappa B p65,reduced intestinal permeability and cell apoptosis,and facilitated the expression of intestinal tight junction proteins in the IUGR colon and the TNF-α-treated cells(P<0.05).Importantly,treatment with tunicamycin or autophagosome-lysosome binding inhibitor chloroquine blocked the positive effects of pterostilbene on inflammatory response,cell apoptosis,and intestinal barrier function in the TNF-α-exposed Caco-2 cells(P<0.05).Conclusion:Pterostilbene mitigates ER stress and promotes autophagic flux,thereby improving colon inflammation and barrier dysfunction in the IUGR piglets and the TNF-α-treated Caco-2 cells.

Intrauterine growth retardation affects liver bile acid metabolism in growing pigs:effects associated with the changes of colonic bile acid derivatives

Background:Intrauterine growth retardation(IUGR)is associated with severely impaired nutrient metabolism and intestinal development of pigs.Our previous study found that IUGR altered intestinal microbiota and metabolites in the colon.However,the consequences of IUGR on bile acid metabolism in pigs remained unclear.The present study aimed to investigate the bile acid metabolism in the liver and the profile of bile acid derivatives in the colon of grow-ing pigs with IUGR using bile acid targeted metabolomics.Furthermore,we determined correlations between colonic microbiota composition and metabolites of IUGR and normal birth weight(NBW)pigs at different growth stages that were 7,21,and 28-day-old,and the average body weight(BW)of 25,50,and 100 kg of the NBW pigs.Results:The results showed that the plasma total bile acid concentration was higher(P<0.05)at the 25 kg BW stage and tended to increase(P=0.08)at 28-day-old in IUGR pigs.The hepatic gene expressions related to bile acid synthe-sis(CYP7A1,CYP27A1,and NTCP)were up-regulated(P<0.05),and the genes related to glucose and lipid metabolism(ATGL,HSL,and PC)were down-regulated(P<0.05)at the 25 kg BW stage in IUGR pigs when compared with the NBW group.Targeted metabolomics analysis showed that 29 bile acids and related compounds were detected in the colon of pigs.The colonic concentrations of dehydrolithocholic acid and apocholic acid were increased(P<0.05),while isodeoxycholic acid and 6,7-diketolithocholic acid were decreased(P<0.05)in IUGR pigs,when compared with the NBW pigs at the 25 kg BW stage.Moreover,Spearman’s correlation analysis revealed that colonic Unclassified_[Mogi-bacteriaceae],Lachnospira,and Slackia abundances were negatively correlated(P<0.05)with dehydrolithocholic acid,as well as the Unclassified_Clostridiaceae abundance with 6,7-diketolithocholic acid at the 25 kg BW stage.Conclusions:These findings suggest that IUGR could affect bile acid and glucolipid metabolism in growing pigs,especially at the 25 kg BW stage,these effects being paralleled by a modification of bile acid derivatives concentra-tions in the colonic content.The plausible links between these modified parameters are discussed.

Effect of Dietary Folic Acid Supplementation on Growth Performance and Hepatic Protein Metabolism in Early-Weaned Intrauterine Growth Retardation Piglets

To investigate the effects of dietary supplementation with folic acid on growth performance, hepatic protein metabolism and serum biochemical indices of early-weaned intrauterine growth retardation （IUGR） piglets, 24 male （Durocx （LandracexYorkshire）） weaned （14-d-old） IUGR piglets were randomly divided into 3 treatments with 8 replicates of 1 piglet per replicate. The piglets in each treatment were fed basal diet supplementation with either 0 （control）, 5 and 10 mg kg^-1 folic acid. The trial lasted for 21 d. Dietary folic acid supplementation reduced average daily feed intake （ADFI） （P〈0.05）. In addition, the average daily gain （ADG） in 10 mg kg^-1 folic acid group was significantly decreased （P〈0.01） and the ratio of feed：gain （F/G） increased slightly （P〉0.05）. Serum folic acid concentration increased （P〈0.01） with increasing folic acid inclusion, however, serum homocysteine concentration decreased significantly （P〈0.01）. Enhanced serum urine nitrogen （SUN） and diminished serum total protein （TP） as well as liver TP content were observed in 10 mg kg^-1 folic acid group （/＇〈0.05）. Furthermore, the relative mRNA expressions of insulin-like growth factor 1 （IGF-1） and mammalian target of rapamycin （m-TOR） in liver were respectively tended to reduce （P=0.06） and significantly downregulated （P〈0.05） in 10 mg kg1 group, in compared with 5 mg kg1 group. However, when compared with control group, folic acid supplementation had no significant effect on the mRNA abundance of IGF- 1 and m-TOR. The results indicated that supplementation with 10 mg kg-I folic acid impaired growth performance and hepatic protein metabolism of early-weaned IUGR piglets while 5 mg kg-~ folic acid enriched diet exerted limited positive effects.

INTRAUTERINE GROWTH RETARDATION AT FULL TERM PREGNANCIES WITH ENDOCRINE FACTORS

Objective To investigate the relationship between intrauterine growth retardation (IUGR) and endocrine parameters so as to assess the effects or the main endocrine ractors on IUGR. The concentrations of growth hormone(GH), insulin, T3, T4 and TSH were measured in umbilical cord blood, amniotic fluid and maternal serum.Methods The samples were collected from 23 pregnant women who were diagnosed as the full term IUGR, 42 normal full term pregnant women with normal infants’ weight were taken as control. Growth hormone and insulin were measured by radioimmunoassay. T3, T4 and TSH were investigated by micro-radioimmunoassay. Results The concentrations of growth hormone, insulin and T4 in umbilical cord blood were lower in IUGR than that in control group(GH4. 63μ/L vs 7. o1μg/L, insulin 1o. 68μIU/ml vs 31. 44μIU/ml, T487. 39nmol/L vs 138. 1onmol/L. P <o. o5, o. o5 and o. o5, respectively). The TSH concentration in umbilical cord blood was higher in IUGR than in control group (1o. 84μmIU/L vs 5. 75μmIU/L, P <o. o1 ). The concentration of growth hormone in maternal serum and the concentration of insulin in amniotic fluid were also lower in IUGR group than in control group(GH 1. 77μg/L vs 2. 74μg/L,P <o. o1, insulin 5. 84μIU/ml vs 15. 64μIU/ml, P <o. o1). Conclusion This study confirms that full term neonates with IUGR are abnormal in endocrine factors. The inadequacy of growth hormone may be one of the causes of IUGR. The relatlve scarcity of growth hormone and insulin seems to be a factor to compromise the fetus’ metabolism. Besides, the early hypothyrosis of infants with IUGR might protect them from unfavorable environment in the uterine.

Dietary dihydroartemisinin supplementation alleviates intestinal inflammatory injury through TLR4/NOD/NF-kB signaling pathway in weaned piglets with intrauterine growth retardation

The aim of present study was to evaluate whether diets supplemented with dihydroartemisinin(DHA)could alleviate intestinal inflammatory injury in weaned piglets with intrauterine growth retardation(IUGR).Twelve normal birth weight(NBW)piglets and 12 piglets with IUGR were fed a basal diet(NBW-CON and IUCR-CON groups),and another 12 piglets with IUGR were fed the basal diet supplemented with DHA at 80 mg/kg(IUGR-DHA group)from 21 to 49 d of age.At 49 d of age,8 piglets with similar body weight in each group were sacrificed.The jejunal and ileal samples were collected for further analysis.The results showed that IUGR impaired intestinal morphology,increased intestinal inflamma-tory response,raised enterocyte apoptosis and reduced enterocyte proliferation and activated trans-membrane toll-like receptor 4(TLR4)/nucleotide-binding and oligomerization domain(NOD)/nuclear factor-kB(NF-kB)signaling pathway.Dihydroartemisinin inclusion ameliorated intestinal morphology,indicated by increased villus height,villus height-to-crypt depth ratio,villus surface area and decreased villus width of piglets with IUGR(P<0.05).Compared with NBW piglets,IUGR piglets supplemented with DHA exhibited higher apoptosis index and caspase-3 expression,and lower proliferation index and proliferating cell nuclear antigen expression in the intestine(P<0.05).Dihydroartemisinin supple-mentation attenuated the intestinal inflammation of piglets with IUGR,indicated by increased concen-trations of intestinal inflammatory cytokines and lipopolysaccharides(P<0.05).In addition,DHA supplementation down-regulated the related mRNA expressions of TLR4/NOD/NF-kB signaling pathway and upregulated mRNA expressions of negative regulators of TLR4 and NOD signaling pathway in the intestine of piglets with IUGR(P<0.05).Piglets in the IUGR-DHA group showed lower protein ex-pressions of TLR4,phosphorylated NF-kB(pNF-kB)inhibitorα,nuclear pNF-kB,and higher protein expression of cytoplasmic pNF-kB in the intestine than those in the IUGR-CON group(P<0.05).In conclusion,DHA supplementation could improve intestinal morphology,regulate enterocyte prolifera-tion and apoptosis,and alleviate intestinal inflammation through TLR4/NOD/NF-kB signaling pathway in weaned piglets with IUGR.

Clinical Study on Effect of Chinese Herbal Medicine for Supplementing Kidney and Qi and Activating Blood Circulation in Treating Intrauterine Growth Retardation of Fetus

Objective: To explore the therapeutic effect and the possible working mechanism in using Chinese herbal medicine (CHM) for supplementing Kidney and Qi, and activating blood circulation in treating intrauterine growth retardation of fetus (IUGR). Methods: Fifty-five cases of IUGR were divided into two groups, 30 cases in the CHM group treated with CHM and the 25 in the control group treated with amino acids. The effect of CHM treatment was observed and compared with that of the control group, normal pregnancy group and non-treated IUGR group. Results: Body weight of the newborns in the CHM was markedly higher than that in the control group. Not only the maternal fundal height (FH) and the abdominal circumference (AC), but also the fetal growth parameters, including biparietal diameter, head circumference (HC), and femur length (FL) in the CHM group increased much faster than those in the control group. After CHM treatment, the maternal serum levels of estriol (E3) and human placental lactogen (hPL) approached to those in the normal pregnancy group, but the control group,in comparison with the normal pregnancy group, was significantly different. The umbilical venous plasma concentration of essential amino acids in both treated groups improved, but the improvement in the CHM group was more significant than that in the control group. No apparent adverse effect of CHM was observed in either mother or fetus.Conclusion: CHM for supplementing Kidney and Qi and activating blood circulation was more effective in improving placental function and enhancing amino acid transportation than amino acid

Resveratrol and its derivative pterostilbene ameliorate intestine injury in intrauterine growth-retarded weanling piglets by modulating redox status and gut microbiota

Background:Intestinal disorder is an important factor contributing to growth lag and high rates of morbidity and mortality of piglets with intrauterine growth retardation(IUGR).Resveratrol(RSV)and its derivative pterostilbene(PT)are natural stilbenes possessing various bioactivities,such as antioxidative and anti-inflammatory effects.This study compared the protective potential of RSV and PT on the intestinal redox status and gut microbiota in weanling piglets with IUGR.Methods:Eighteen male piglets of normal body weight(NBW)and 54 same-sex IUGR piglets were chosen according to their birth and weaning weights.The NBW piglets accepted a basal diet,while the IUGR piglets were allotted to one of three groups according to their body weight at weaning and received a basal diet,an RSV-supplemented diet(300 mg/kg),or a PT-supplemented diet(300 mg/kg),respectively.Results:Compared with IUGR piglets,both RSV and PT improved the IUGR-associated decrease in jejunal villus height and increases in plasma diamine oxidase activity and D-lactate level and jejunal apoptosis of piglets(P<0.05).Administering RSV and PT also enhanced jejunal superoxide dismutase activity and the mRNA and protein expression of superoxide dismutase 2 of IUGR piglets by promoting nuclear factor erythroid 2-related factor 2(Nrf2)nuclear translocation(P<0.05).Comparatively,PT was more effective than RSV in elevating the villus height/crypt depth ratio and occludin mRNA and protein levels in the jejunum of IUGR piglets(P<0.05).PT was also superior to RSV in increasing Nrf2 nuclear translocation and inhibiting malondialdehyde accumulation in the jejunum of IUGR piglets(P<0.05).Additionally,RSV modulated the composition of cecal microbiota of IUGR piglets,as evidenced by increasing the prevalence of the phylum Bacteroidetes and the genera Prevotella,Faecalibacterium,and Parabacteroides and inhibiting the growth of the phylum Proteobacteria and its genera Escherichia and Actinobacillus(P<0.05).Moreover,RSV significantly increased the butyrate concentration in the cecum of IUGR piglets(P<0.05).Conclusion:PT is more potent than RSV to prevent intestinal oxidative stress,while RSV has a stronger capacity to regulate gut microbiota compared to PT.

Effects of Maternal Folic Acid Supplementation on Hepatic Apoptosis-Related Gene Expressions between Intrauterine Growth Restriction and Normal Body Weight Piglets

Intrauterine growth retardation （IU- GR） causes significantly negative effects on the meth- ylation status of genes related to cell apoptosis com- pared with normal body weight （NBW） piglets. Thus, the objective of the present study was to exam- ine the effects of maternal dietary folic acid supple- mentation on genes expression profile for hepatic ap- optosis in IUGR and NBW piglets. Twenty four York- shire gilts were allocated randomly to one of the two diets ： control （ C, folic acid 1.3 mg/kg） or folic acid supplementation （ FS, folic acid 30 mg/kg） after mat- ing. Gene expressions in liver samples were deter- mined and revealed that the mRNA expressions of p53 ,BCL-2 associated X protein （Bax）, and Cyclin- dependent kinase inhibitor 1A （CDKN1A） were up-regulated in IUGR piglets compared with NBW pig- lets fed C diets,but could be reversed by maternal fo- lic acid supplementation. The expressions of vascular endothelial growth factor （VEGF）, Serine-protein Ki- nase-Ataxia Telangiectasia Mutated （ATM） ,and Cad- herin-associated protein-beta-catenin 1 （ CTNNB1 ） were influenced by maternal folic acid supplementa- tion significantly, but were not influenced by birth weight. Expression of p53 binding protein-MDM-2 （ MDM-2 ） remained unchanged. In conclusion, these results demonstrated that maternal folic acid supple- mentation could exert positive effects on genes related to apoptosis in IUGR and NBW piglets, which might facilitate their postnatal health and growth perform- alice.

Restored intestinal integrity,nutrients transporters,energy metabolism,antioxidative capacity and decreased harmful microbiota were associated with IUGR piglet's catch-up growth before weanling

Background:Intrauterine growth restriction(IUGR)is a major inducer of higher morbidity and mortality in the pig industry and catch-up growth(CUG)before weanling could significantly restore this negative influence.But there was limited knowledge about the underlying mechanism of CUG occurrence.Methods:Eighty litters of newborn piglets were divided into normal birth weight(NBW)and IUGR groups according to birth weight.At 26 d,those piglets with IUGR but over average body weight of eighty litters of weaned piglets were considered as CUG,and the piglets with IUGR still below average body weight were considered as NCUG.This study was conducted to systemically compare the intestinal difference among NBW,CUG and NCUG weaned piglets considering the crucial role of the intestine for piglet growth.Results:The results indicated that the m RNA expression of nutrients(amino acids,glucose,and fatty acids)transporters,and mitochondrial electron transport chain(ETC)I were upregulated in CUG piglets'gut with improved morphology compared with those NCUG,as well as the ratio of P-AMPK/AMPK protein expression which is the indicator of energy metabolism.Meanwhile,CUG piglet's gut showed higher antioxidative capacity with increased SOD and GSHPx activity,decreased MDA levels,as well as higher m RNA expressions of Nrf2,Keap1,SOD,and GSH-Px.Furthermore,inflammatory parameters including TNF-α,IL-1β,IL-6,and IL-12 factors,and the activation of MAPK and NF-κB signaling pathways were significantly elevated in the NCUG intestine,while the protein expression of ZO-1,Occludin and Claudin-1 was reduced.The alpha diversity of fecal microbiota was higher in CUG piglets in contrast with NCUG piglets,and the increased beneficial bacteria and decreased pathogenic bacteria was also observed in CUG piglets.Conclusions:CUG piglet's intestine showed comprehensive restoration including higher nutrients transport,energy metabolism,antioxidant capacity,and intestinal physical barrier,while lower oxidative stress,inflammatory response,and pathogenic microbiota.

Study on the Effects of ＂Foshousan＂ Plus Danshen in Preventing IUGR Rats with Passive Smoking from Peroxidation in Erythrocyt

The pregnant rats with cigarette smoking exposure were used as the animal model for studying the pathogenesis of IUGR. The mechanism of preventing intrauterine growth retardation with 'Foshousan' plus Danshen by protecting erythrocytes against lipid peroxidation damage was also preliminarily discussed.The erythrocyte membrane lipid peroxide content, erythrocyte SOD activities were detected by ultraviolet split-beam spectrophotometry and chemiluminescence technique. The results showed that the erythrocyte MDA levels and ratio of abnormal erythrocyte in model group were significantly higher than those in control group and treated group, and the fetal mean birth weight, erythrocyte SOD levels were all statistically significantly lower than those in control group and treated group. The fetal birth weight showed a significant positive correlation with SOD levels and a significant negative correlation with MDA level. The results suggest 'Foshousan' plus Danshen can protect erythrocytes against lipid peroxidation damage, thus conserving the normal form, structure and function of erythrocytes, improving the uteroplacental blood flow and thereby increasing the fetal mean birth weight.

The Effects of "Huoxuequyu" Decoction on Lipid Peroxidation in IUGR Rats

The rats exposed to passive smoking were used as the animal model to further study the pathogenesis of IUGR and the effects of ' Huoxuequyu' decoction. The results showed that the mean birth-weight of fetus and SOD activity of placenta and plasma in model rats were significantly reduced, and LPO levels of placenta and plasma were significantly increased. We found that the mean birthweight and SOD activity in treated group were higher than that in model group,and LPO levels of placenta and plasma were lower in treated group than that in model group and there was no difference in SOD activity and LPO levels of placenta and plasma as well as the mean birthweight between treated group and control group, indicating that 'Huoxuequyu' decoction can protect placenta and plasma against lipid peroxidation, and improve the blood circulation of uteroplacenta, thus contributing to the increase of the mean birth-weight.

THE EFFECTS OF ZINC SUPPLEMENTATION TO CHINESE RURAL PREGNANT WOMEN AND THEIR PREGNANCY OUTCOME

Objective: TO assess the effects of zinc supplementation to Chinese rural pregnant with cereal-based diet and their pregnancy outcome. Methods A total of 156 women were given randomly in double blind 5mg/d (group A, GPA, n=37),10mg/d (group B, GPB, n=40), 30mg (group C, GpC, n = 39) Of zinc supplement or placebo (group D, GpD, n = 40 ) starting from, the first visit at the prenatal clinic (average 9-week gestation ) till the delivery. Results Mean dietary zinc intake of these women was about 10mg/d, and more than 65% of dietary zinc were taken from cereals, grains and vegetables. Serum zinc concentrations at 25-and 32-gestation in GpC and in the umbilical cord blood of their babies at birth significantly increased. Infants in GpC had a significantly heavier birth weight (283. 079, P = 0. 016 ) and larger head circumference(0. 63cm, P = 0. 035) than infants in GpD. The incidence of low birth weight, preterm and intrauterine growth retardation infants was significantly reduced in GpC. As compared to GpD (38. 34 ± 1. 34 weeks), the duration of gestation in GpD(39. 12± 1. 11 weeks) was prolonged (P= 0. 032). Serum zinc concentration at 25-weeks gestation was negatively correlated with duration of gestation (P = 0. 009 ). Apgar score of the infants in GpC was significantly higher than that in GpD, and serum zinc concentration at 32-weeks gestation was positively associated with infant Apgar score (P = 0. 015 ). Conclusion Zinc supplementation in 30mg/d to Chinese rural women during pregnancy could significantly improve their pregnancy outcome. Both 5mg/d and 10mg/d of zinc supplementation had no detected benefits to them.

Epigenetics in neonatal diseases

Objective To review the role of epigenetic regulation in neonatal diseases and better understand Barker＇s ＂fetal origins of adult disease hypothesis＂.Data sources The data cited in this review were mainly obtained from the articles published in Medline/PubMed between January 1953 and December 2009.Study selection Articles associated with epigenetics and neonatal diseases were selected.Results There is a wealth of epidemiological evidence that lower birth weight is strongly correlated with an increased risk of adult diseases, such as type 2 diabetes mellitus, hypertension, and cardiovascular disease. This phenomenon of fetal origins of adult disease is strongly associated with fetal insults to epigenetic modifications of genes. A potential role of epigenetic modifications in congenital disorders, transient neonatal diabetes mellitus （TNDM）, intrauterine growth retardation （IUGR）, and persistent pulmonary hypertension of the newborn （PPHN） have been studied.Conclusions Acknowledgment of the role of these epigenetic modifications in neonatal diseases would be conducive to better understanding the pathogenesis of these diseases, and provide new insight for improved treatment and prevention of later adult diseases.