Dietary supplementation with garcinol during late gestation alleviates disorders of bile acid metabolism and improves the performance of sows and newborn piglets.

The present study was conducted to evaluate the effects of dietary garcinol supplementation during late gestation on bile acid metabolism and performance of sows. Sixty sows (Duroc × Yorkshire × Landrace; second- or third-parity; n = 20) with disorder of bile acid metabolism were randomly divided into three groups: control diet (CON; basal diet), basal diet with 200 mg garcinol (Low Gar), and basal diet with 600 mg garcinol (High Gar) per kg of feed. The body weight (BW); backfat thickness and litter size of the sows; and birth weight, weaning weight, and mortality of piglets were recorded. Sows' blood was collected for the measurements of hematological parameters and antioxidative and immune indexes, and indicators related to bile acid metabolism, respectively. The colostrum and fecal samples of the sows were also collected for analysis of colostrum composition and apparent total tract nutrient digestibility. Garcinol had no effect on the BW and backfat thickness of the sows but significantly decreased the mortality and number of weak litter (P < 0.05). Moreover, the white blood cell counts, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) activity in the plasma of the sows were increased more significantly (P < 0.05) in the garcinol groups than that in the CON group, whereas the malondialdehyde (MDA) content was decreased (P < 0.05). Dietary supplementation with garcinol significantly reduced TBA concentrations (P < 0.05). The content of immunoglobulin A (IgA) and immunoglobulin G (IgG) in the plasma and in colostrum of sows were increased more significantly (P < 0.05) in the garcinol groups than that in the CON group. In addition, dry matter (DM), Ash, and EE in the colostrum were similar between groups (P > 0.05), whereas the garcinol significantly increased the crude protein (CP) in the colostrum. The apparent total tract nutrient digestibility was similar between treatments. Garcinol treatment induced a gradually decreased (P > 0.05) the expression of genes involved in BA synthesis (CYP7A1, CYP8B1), BA uptake (NTCP, OATP1A2), BA secretion (BSEP and MRP2), BA detoxification (SULT2A1), and BA efflux into the blood circulation (OSTß). Collectively, this study indicates that sows fed with garcinol in late gestation showed relieved bile acid metabolism disorder and improved sows performance, antioxidative status, colostrum protein content, showing promise in natural plant extract nutrition for sows with disorder of bile acid metabolism.

The elevated maternal serum bile acid (BA) levels in late gestation leads to accumulation of BA in fetal tissues, and thus increases the risk of fetal mortality and metabolic disease of offspring. It has confirmed that BA disordered and oxidative damage are intimately related. Thus, studies about alleviating oxidative stress and facilitating BA metabolism in pregnant sows can be relevant. As an excellent antioxidative plant extract, garcinol has been widely used in dietary supplementation of rodents; however, the effect of dietary supplementation with garcinol on the bile acids disorders of sows in late gestation has rarely been reported. The present study provides the first evidence that dietary supplementation with garcinol during late gestation improved maternal BA metabolism of sows with disorder of BA metabolism, as well as the health and antioxidative status, colostrum protein content, showing promise in natural plant extract nutrition for sows with disorder of bile acid metabolism.

Effects of garcinol supplementation on the performance, egg quality, and intestinal health of laying hens in the late laying period.

The problem of rapid decline in egg production performance and poor egg quality is a key obstacle to improving the economic benefits of laying hens. Garcinol is an antioxidant polyphenol plant extract that has multiple physiological functions. Diets with the appropriate amount of garcinol might be able to improve the performance traits and health of late laying hens. Therefore, this study was conducted to evaluate the utilization of garcinol in late laying hens. A total of 400 healthy 59-wk-old Tingfen No. 6 hens were randomly allocated into 4 dietary treatment groups and fed a basal diet supplemented with 0, 100, 300, and 500 mg/kg garcinol for 12 wk, denoted the Con, LG, MG, and HG groups, respectively. The results showed that the addition of garcinol in the diet tended to increase the egg production rate compared with that of the control group (P = 0.080), while the average egg weight was significantly lower (P < 0.05) during the whole period of the experiment. The results showed that MG group hens had higher egg quality and strengthened antioxidant capacity in their serum (P < 0.05). Moreover, the laying hens in the MG group had significantly decreased crypt depth (CD) and increased villus height (VH) in the jejunum and ileum (P < 0.05), as well as an increased ratio of VH to CD (P < 0.05) and increased expression levels of Occludin (P < 0.05) and Claudin-2 (P < 0.05) in the jejunum to improve intestinal barrier function. In addition, dietary supplementation with garcinol influenced the cecal microbiota of laying hens, which was characterized by changes in the microbial community composition, including increased abundances of Firmicutes, Romboutsia, and Ruminococcus torques. In conclusion, dietary 300 mg/kg garcinol supplementation could increase the egg production and egg quality of late laying hens, which may be attributed to the antioxidant effects of garcinol and the improvement of intestinal morphology and epithelial barrier function as well as the regulation of mucosal immune status by altering microbial composition.

Effects of different processing techniques of palm kernel cake on processing quality of pellet feed, nutrient digestibility, and intestinal microbiota of pigs.

The present study was conducted to investigate the effects of extrusion, fermentation, and enzymolysis of palm kernel cake on processing quality of pellet feed, nutrient digestibility, and intestinal microbiota of pigs. First, the pretreatment parameters of extrusion, enzymolysis, and fermentation of palm kernel cake were optimized. Then, PKC after three processing techniques were used to prepare pellet feed. A total of 160 crossbred piglets (Duroc × Landrace × Yorkshire) with an average body weight of 28 ±â€…0.5 kg were used in an 8-wk feeding experiment. Pigs were randomly assigned to five treatments with four replicates per treatment and eight pigs per replicates. The five experimental groups were as follows: basal diet group (whole corn-soybean meal), 10% PKC group (PKC), 10% extrusion PKC group (PPKC), 10% enzymolysis PKC group (EPKC), and 10% fermented PKC group (FPKC), respectively. At the end of the experiment, four pigs from each treatment (randomly collected one pig per pen) were sacrificed by administering a pentobarbital overdose, the gut and blood samples were collected for the quantification analysis of microbiota, hematological parameters, and apparent total tract nutrient digestibility. The results showed that all three processing techniques significantly decreased the contents of crude fiber of PKC (P < 0.01), pulverization rate (P < 0.01), powder content (P < 0.01), and increased the hardness and gelatinization starch of pellet feed (P < 0.05) compared to PKC group. In addition, PPKC significantly improved the dry matter, crude protein, and ether extract content, blood indices and average daily feed intake compared to PKC group (P < 0.01), while the parameters were similar among FPKC, EPKC, and control group (P > 0.01). Furthermore, all three processing techniques significantly increased the Lactobacillus and decreased the Escherichia levels in feces or gut compared to PKC. Collectively, extrusion, fermentation, and enzymolysis of PKC had positively enhanced the pellet quality, growth performance, nutrient digestibility, and gut microbiota, extrusion exhibited a superior feeding effect compared to fermentation and enzymolysis.

Palm kernel cake (PKC) has lower nutritional value compared with soybean meal, cottonseed meal, and rapeseed meal, but its cost advantage is great, and it has been gradually used in the ruminant feeding. Due to its high crude fiber content, the processing technique applied to the PKC has a significant impact on its effectiveness. However, the different processing techniques of PKC on pellet quality, and performance of pigs have been poorly reported. The present study was conducted to investigate the effects of extrusion, fermentation, and enzymolysis pretreatment of PKC on processing quality of pellet feed, nutrient digestibility (in vivo), and intestinal microbiota of growing­finishing pigs. This study provides the optimal processing parameters of the three processing techniques, and demonstrated that PKC after processing could significantly improve the pellet quality, performance, and intestinal microbiota of growing­finishing pigs, while extrusion exhibited a superior feeding effect compared to fermentation and enzymolysis.

Comprehensive Proteome and Acetyl-Proteome Atlas Reveals Hepatic Lipid Metabolism in Layer Hens with Fatty Liver Hemorrhagic Syndrome.

The feeding of high-energy and low-protein diets often induces fatty liver hemorrhagic syndrome (FLHS) in laying hens. However, the mechanism of hepatic fat accumulation in hens with FLHS remains uncertain. In this research, a comprehensive hepatic proteome and acetyl-proteome analysis was performed in both normal and FLHS-affected hens. The results indicated that the upregulated proteins were primarily associated with fat digestion and absorption, the biosynthesis of unsaturated fatty acids, and glycerophospholipid metabolism, while the downregulated proteins were mainly related to bile secretion and amino acid metabolism. Furthermore, the significant acetylated proteins were largely involved in ribosome and fatty acid degradation, and the PPAR signaling pathway, while the significant deacetylated proteins were related to valine, leucine, and isoleucine degradation in laying hens with FLHS. Overall, these results demonstrate that acetylation inhibits hepatic fatty acid oxidation and transport in hens with FLHS, and mainly exerts its effects by affecting protein activity rather than expression. This study provides new nutritional regulation options to alleviate FLHS in laying hens.

Effects of different processing techniques of broken rice on processing quality of pellet feed, nutrient digestibility, and gut microbiota of weaned piglets.

The present study was conducted to assess the effect of different processing techniques of broken rice on processing quality of pellet feed, growth performance, nutrient digestibility, blood biochemical parameters, and fecal microbiota of weaned piglets. A total of 400 crossbred piglets (Duroc × Landrace × Yorkshire) with a mean initial body weight (BW) of 7.24 ±â€…0.52 kg were used in a 28-d experiment. Piglets were randomly distributed to one of 4 treatment and 10 replicate pens per treatment, with 10 piglets per pen. The dietary treatments were as follows: CON, corn as the main cereal type in the dietary; BR, 70% of the corn replaced by broken rice; ETBR, 70% of the corn replaced by extruded broken rice; EPBR, 70% of the corn replaced by expanded broken rice. Extruded broken rice and expanded broken rice supplementation significantly (P < 0.05) increased hardness, pellet durability index, crispness, and starch gelatinization degree. Extruded broken rice and expanded broken rice generated a higher (P < 0.05) average daily feed intake, increased (P < 0.05) average daily gain, decreased (P < 0.05) feed conversion ratio, and lowered (P < 0.05) the diarrhea rate. Piglets fed extruded broken rice displayed high apparent total tract digestibility levels of dry matter (P < 0.05), gross energy (P < 0.05), crude protein (P < 0.05), and organic matter (P < 0.05). In addition, extruded broken rice and expanded broken rice supplementation had increased Lactobacillus and Bifidobacterium levels in gut, whereas a lower abundance of the potential pathogens Clostridium_sensu_strictio_1 and Streptococcus was observed. Dietary supplementation of extruded broken rice and expanded broken rice failed to show significant effects on blood biochemical parameters. Combined, 70% corn substituted with broken rice failed to show significant effects. Collectively, extruded broken rice and expanded broken rice supplementation had positively enhanced the pellet quality, growth performance, nutrient digestibility, and gut microbiota of weaned piglets.

Weaned piglets represent a critical phase in animal husbandry, and with the rising demand for meat, the consumption of animal feed has surged. Corn, a vital constituent of animal feed, has been consumed at an accelerated pace. In this regard, the use of broken rice as an alternative to corn is a feasible solution. Nevertheless, due to the incomplete development of piglets' bodies, higher quality feed is necessary. The processing technique applied to the feed has a significant impact on its effectiveness. Thus, we experimented to assess the effect of different processing techniques on the feed efficiency of weaning piglets, substituting corn with broken rice, extruded broken rice, and expanded broken rice. The study results revealed that the application of extruded and expanded broken rice improved the feed pellet quality, growth performance, nutrient digestibility, and gut microbiota of weaned piglets. Furthermore, extruded broken rice exhibited a superior feeding effect compared to expanded broken rice.

Garcinol Promotes the Formation of Slow-Twitch Muscle Fibers by Inhibiting p300-Dependent Acetylation of PGC-1α.

The conversion of skeletal muscle fiber from fast-twitch to slow-twitch is crucial for sustained contractile and stretchable events, energy homeostasis, and anti-fatigue ability. The purpose of our study was to explore the mechanism and effects of garcinol on the regulation of skeletal muscle fiber type transformation. Forty 21-day-old male C57/BL6J mice (n = 10/diet) were fed a control diet or a control diet plus garcinol at 100 mg/kg (Low Gar), 300 mg/kg (Mid Gar), or 500 mg/kg (High Gar) for 12 weeks. The tibialis anterior (TA) and soleus muscles were collected for protein and immunoprecipitation analyses. Dietary garcinol significantly downregulated (p < 0.05) fast myosin heavy chain (MyHC) expression and upregulated (p < 0.05) slow MyHC expression in the TA and soleus muscles. Garcinol significantly increased (p < 0.05) the activity of peroxisome proliferator-activated receptor gamma co-activator 1α (PGC-1α) and markedly decreased (p < 0.05) the acetylation of PGC-1α. In vitro and in vivo experiments showed that garcinol decreased (p < 0.05) lactate dehydrogenase activity and increased (p < 0.05) the activities of malate dehydrogenase and succinic dehydrogenase. In addition, the results of C2C12 myotubes showed that garcinol treatment increased (p < 0.05) the transformation of glycolytic muscle fiber to oxidative muscle fiber by 45.9%. Garcinol treatment and p300 interference reduced (p < 0.05) the expression of fast MyHC but increased (p < 0.05) the expression of slow MyHC in vitro. Moreover, the acetylation of PGC-1α was significantly decreased (p < 0.05). Garcinol promotes the transformation of skeletal muscle fibers from the fast-glycolytic type to the slow-oxidative type through the p300/PGC-1α signaling pathway in C2C12 myotubes.

Dietary embelin supplementation during mid-to-late gestation improves performance and maternal-fetal glucose metabolism of pigs.

This study aimed to evaluate the effects of dietary embelin supplementation during late gestation (from days 60 to 110) on performance and maternal-fetal glucose metabolism of pigs. Sixty sows (Duroc × Yorkshire × Landrace; parity = 1.68 ±â€…0.03; N = 20) were randomly divided into three gestation (day 60 of pregnancy) treatments, Control pigs (CON) were fed a basal diet, and the other animals were fed a basal diet supplemented with 200 or 600 mg/kg embelin per kg of feed. The body weight, backfat thickness and litter size of the sows, and birth weight and mortality of piglets were recorded. Sows' blood and piglets' umbilical cord blood were collected for the measurements of hematological parameters and anti-oxidative and immune indexes, and maternal-fetal glucose metabolism parameters, respectively. The colostrum and milk and fecal samples of the sows were also collected for analysis of milk composition and apparent total tract nutrient digestibility. Dietary embelin had no effect on the BW and backfat thickness of the sows but significantly increased the birth weight of piglets (P < 0.05) and decreased the mortality (P < 0.05). Moreover, the white blood cell counts (day 90), neutrophil count and mean cell hemoglobin (day 110), total anti-oxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) content of the sows were increased significantly (P < 0.05) in the embelin groups than that in the CON group, whereas the malondialdehyde (MDA) content was decreased (P < 0.05). Embelin significantly increased immunoglobulin A (IgA) and immunoglobulin G (IgG) content in plasma of piglets as well as those in colostrum and milk of sows than the CON treatment (P < 0.05). In addition, dry matter, ash, and ether extract in the colostrum were similar between groups (P > 0.05), whereas the embelin significantly increased the crude protein in the milk. The apparent total tract nutrient digestibility was similar between treatments (P > 0.05). The embelin treatment significantly increased the glucose levels and lactate dehydrogenase B (LDHB) activity in sows plasma, and decreased the lactate levels in both sows and fetuses plasma (P < 0.05). Collectively, this study indicates that sows fed with embelin in mid-to-late gestation showed improved maternal health and anti-oxidative status, milk protein content, and maternal-fetal glucose metabolism, showing promise in natural plant extract nutrition for sows.

Abnormal glucose metabolism in sows in late gestation can lead to incapacity of sow production, and even reproductive disorders. It has been confirmed that inefficient glucose utilization and oxidative damage are intimately related. Thus, studies about alleviating oxidative stress and facilitating glucose metabolism in pregnant sows can be relevant. As an excellent anti-oxidative plant extract, embelin has been widely used in dietary supplementation of rodents, however, the effect of dietary supplementation with embelin on the performance of sows and newborn piglets, as well as on the glucose metabolism has rarely been reported. The present study provides the first evidence that dietary supplementation with embelin during mid-to-late gestation improved maternal immune and oxidative status, the milk quality as well as the glucose metabolism of both sows and piglets, suggesting that embelin may be a promising natural plant extractive of nutrition for sows especially during mid-to-late pregnancy and lactation.

Embelin alleviates weaned piglets intestinal inflammation and barrier dysfunction via PCAF/NF-κB signaling pathway in intestinal epithelial cells.

BACKGROUND: Intestinal barrier plays key roles in maintaining intestinal homeostasis. Inflammation damage can severely destroy the intestinal integrity of mammals. This study was conducted to investigate the protective effects of embelin and its molecular mechanisms on intestinal inflammation in a porcine model. One hundred sixty 21-day-old castrated weaned pigs (Duroc × Landrace × Yorkshire, average initial body weight was 7.05 ± 0.28 kg, equal numbers of castrated males and females) were allotted to four groups and fed with a basal diet or a basal diet containing 200, 400, or 600 mg embelin/kg for 28 d. The growth performance, intestinal inflammatory cytokines, morphology of jejunum and ileum, tight junctions in the intestinal mucosa of piglets were tested. IPEC-1 cells with overexpression of P300/CBP associating factor (PCAF) were treated with embelin, the activity of PCAF and acetylation of nuclear factor-κB (NF-κB) were analyzed to determine the effect of embelin on PCAF/NF-κB pathway in vitro. RESULTS: The results showed that embelin decreased (P < 0.05) serum D-lactate and diamine oxidase (DAO) levels, and enhanced the expression of ZO-1, occludin and claudin-1 protein in jejunum and ileum. Moreover, the expression levels of critical inflammation molecules (interleukin-1ß, interleukin-6, tumor necrosis factor-α, and NF-κB) were down-regulated (P < 0.05) by embelin in jejunal and ileal mucosa. Meanwhile, the activity of PCAF were down-regulated (P < 0.05) by embelin. Importantly, transfection of PCAF siRNAs to IPEC-1 cell decreased NF-κB activities; embelin treatment downregulated (P < 0.05) the acetylation and activities of NF-κB by 31.7%-74.6% in IPEC-1 cells with overexpression of PCAF. CONCLUSIONS: These results suggested that embelin ameliorates intestinal inflammation in weaned pigs, which might be mediated by suppressing the PCAF/NF-κB signaling pathway.

Dietary Se-Enriched Cardamine enshiensis Supplementation Alleviates Transport-Stress-Induced Body Weight Loss, Anti-Oxidative Capacity and Meat Quality Impairments of Broilers.

The aim of this experiment was to explore the effects of a new selenium (Se) source from Se-enriched Cardamine enshiensis (SeCe) on body weight loss, anti-oxidative capacity and meat quality of broilers under transport stress. A total of 240 one-day-old ROSS 308 broilers were allotted into four treatments with six replicate cages and 10 birds per cage using a 2 × 2 factorial design. The four groups were as follows: (1) Na2SeO3-NTS group, dietary 0.3 mg/kg Se from Na2SeO3 without transport stress, (2) SeCe-NTS group, dietary 0.3 mg/kg Se from SeCe without transport stress, (3) Na2SeO3-TS group, dietary 0.3 mg/kg Se from Na2SeO3 with transport stress, and (4) SeCe-TS group, dietary 0.3 mg/kg Se from SeCe with transport stress. After a 42 d feeding period, the broilers were transported by a lorry or kept in the original cages for 3 h, respectively. The results showed that dietary SeCe supplementation alleviated transport-stress-induced body weight loss and hepatomegaly of the broilers compared with the broilers fed Na2SeO3 diets (p < 0.05). Furthermore, dietary SeCe supplementation increased the concentrations of plasma total protein and glucose, and decreased the activities of aspartate aminotransferase and alanine aminotransferase of the broilers under transport stress (p < 0.05). Dietary SeCe supplementation also enhanced the anti-oxidative capacity and meat quality in the breast and thigh muscles of the broilers under transport stress (p < 0.05). In summary, compared with Na2SeO3, dietary SeCe supplementation alleviates transport-stress-induced body weight loss, anti-oxidative capacity and meat quality impairments of broilers.

Targeting gut microbiota-derived butyrate improves hepatic gluconeogenesis through the cAMP-PKA-GCN5 pathway in late pregnant sows.

Short chain fatty acids (SCFAs) produced by gut microbiota affected hepatic glucose metabolism via the gut-liver axis. The present study aimed to investigate the effects of butyrate produced by gut microbiota on hepatic gluconeogenesis in late-pregnancy sows. A total of 240 primiparous sows in late pregnancy were tested for blood glucose using a glucose meter before feeding and grouped according to their blood glucose level as follows: 0-3.0 mmol L-1 (low blood glucose group, LG group) and 3.1-5.0 mmol L-1 (normal blood glucose group, NG group). Colonic SCFAs and microbiota, SCFAs in the portal vein and liver, and acetylation and phosphorylation levels in the liver samples were analyzed. Hepatocytes from pregnant sows were examined for the effect of butyrate on hepatic glucose gluconeogenesis. In vivo experiments showed that the reproductive performance, serum glucose metabolism index, colonic butyrate and butyrate-producing bacteria decreased in the LG group compared with the NG group. Correlation analysis found a positive correlation among colonic butyrate, butyrate-producing bacteria and the serum glucose metabolism index. Moreover, the hepatic cAMP concentration, PKA activity, GCN5 phosphorylation, and the expression of G6P and PEPCK were decreased and PGC1-α acetylation was increased in the LG group compared with the NG group. In vitro, sodium butyrate significantly stimulated the cAMP concentration, PKA activity, GCN5 phosphorylation, and the expression of G6P and PEPCK and inhibited PGC-1α acetylation in the LG group of hepatocytes from late-pregnancy sows. Interestingly, another in vivo experiment showed that dietary 1-kestose, a natural regulator of gut bacteria, significantly increased butyrate and butyrate-producing bacteria, and improved the reproductive performance and serum glucose metabolism index in late-pregnancy sows. Taken together, we found that targeting gut microbiota-derived butyrate could improve hepatic gluconeogenesis through the cAMP-PKA-GCN5 pathway in late-pregnancy sows.

Acetylation of lactate dehydrogenase B drives NAFLD progression by impairing lactate clearance.

BACKGROUND & AIMS: Lactate has recently been reported to accumulate in the livers of patients progressing from simple steatosis to non-alcoholic steatohepatitis (NASH). However, the underlying mechanism(s) of lactate accumulation and the role of lactate in the progression of non-alcoholic fatty liver disease (NAFLD) are essentially unknown. METHODS: We compared the acetylome in liver samples taken from healthy individuals, patients with simple steatosis and patients with NASH to identify potential targets of acetylation with a role in lactate metabolism. Interactions between the acetylated target and acetyltransferases were measured in multiple cell lines. An acetyltransferase inhibitor was injected into high-fat diet (HFD)-fed mice to determine the role of lactate on NAFLD progression in vivo. RESULTS: Hyperacetylation of lactate dehydrogenase B (LDHB) was found to be associated with lactate accumulation in NAFL and NASH livers in humans and mice. P300/CBP-associated factor (PCAF)-mediated acetylation of LDHB K82 was found to significantly decrease LDHB activity and impair hepatic lactate clearance, resulting in lactate accumulation. Acetylated LDHB induced lactate accumulation which exacerbated lipid deposition and inflammatory responses by activating histone hyperacetylation in HFD-induced NASH. The administration of embelin, a PCAF inhibitor, and the generation of an acetylation-deficient mutant of LDHB ameliorated NASH. CONCLUSION: PCAF-dependent LDHB acetylation plays a key role in hepatic lipid accumulation and inflammatory responses by impairing lactate clearance; this process might be a potential therapeutic target for the treatment of NASH. LAY SUMMARY: Lactate is known to accumulate in the livers of patients during the progression of non-alcoholic fatty liver disease (NAFLD); however, the underlying mechanism(s) of this accumulation and its importance in disease progression are unknown. Herein, we show that the acetylation of an enzyme involved in lactate metabolism leads to impaired lactate clearance and exacerbates NAFLD progression.

Garcinol promotes hepatic gluconeogenesis by inhibiting P300/CBP-associated factor in late-pregnant sows.

Disorder of hepatic glucose metabolism is the characteristic of late-pregnant sows. The purpose of our study was to look into the mechanism of garcinol on the improvement of hepatic gluconeogenic enzyme in late-pregnant sows. Thirty second- and third-parity sows (Duroc × Yorkshire × Landrace, n 10/diet) were fed a basal diet (control) or that diet supplemented with 100 mg/kg (Low Gar) or 500 mg/kg (High Gar) garcinol from day 90 of gestation to the end of farrowing. The livers were processed to measure enzymatic activity. Hepatocytes from pregnant sows were transfected with P300/CBP-associating factor (PCAF) small interfering RNA (siRNA) or treated with garcinol. Dietary garcinol had no effect on average daily feed intake, body weight (BW), backfat and BW gain of late-pregnant sows. Garcinol promoted plasma glucose levels in pregnant sows and newborn piglets. Garcinol up-regulated hepatic gluconeogenic enzyme expression and decreased PCAF activity. Garcinol had no effect on the expression of PPAR-γ co-activator 1α (PGC-1α) and Forkhead box O1 (FOXO1) but significantly increased their activity and decreased their acetylation in late-pregnant sows. Transfection of PCAF siRNA to hepatocytes of pregnant sows increased PGC-1α and FOXO1 activities. Furthermore, in hepatocytes of pregnant sows, garcinol treatment also up-regulated the activities of PGC-1α and FOXO1 and inhibited the acetylation of PGC-1α and FOXO1. Garcinol improves hepatic gluconeogenic enzyme expression in late-pregnant sows, and this may be due to the mechanism of down-regulating the acetylation of PGC-1α and FOXO1 induced by PCAF in isolated hepatocytes.

Acetylation of Phenylalanine Hydroxylase and Tryptophan 2,3-Dioxygenase Alters Hepatic Aromatic Amino Acid Metabolism in Weaned Piglets.

Weaning significantly alters hepatic aromatic amino acid (AAA) metabolism and physiological functions. However, less is known about the regulating mechanism of hepatic AAA metabolism after weaning. A total of 200 21-day-old piglets (Duroc × Landrace) were assigned randomly to the control group and the weaning group. In this study, weaning significantly decreased the concentration of phenylalanine, tryptophan, and tyrosine in piglet livers (p < 0.05). Additionally, through the detection of liver AAA metabolites and metabolic enzyme activity, it was observed that hepatic tryptophan catabolism was enhanced, while that of phenylalanine was weakened (p < 0.05). Intriguingly, acetyl-proteome profiling of liver from weaned piglets showed that weaning exacerbated the acetylation of phenylalanine hydroxylase (PAH) and the deacetylation of tryptophan 2,3-dioxygenase (TDO). Analysis of PAH and TDO acetylation in Chang liver cells showed that acetylation decreased the PAH activity, while deacetylation increased the TDO activity (p < 0.05). Furthermore, metabolites of AAAs and the acetylation statuses of PAH and TDO in primary hepatocytes from weaned piglets were consistent with the results in vivo. These findings indicated that weaning altered the PAH and TDO activity by affecting the acetylation state of the enzyme in piglets'' livers. Lysine acetylation may be a potential regulatory mechanism for AAA metabolism in response to weaning.

Dietary garcinol supplementation improves diarrhea and intestinal barrier function associated with its modulation of gut microbiota in weaned piglets.

BACKGROUND: The effects of dietary garcinol on diarrhea and intestinal barrier function associated with its modulation of gut microbiota in weaned piglets were investigated. METHOD: One hundred forty four weaned piglets (Duroc × Yorkshire × Landrace) from 16 pens (9 piglets per pen) were randomly divided into four treatment groups: controls (CON) or those supplemented with 200 mg/kg (LOW), 400 mg/kg (MID), or 600 mg/kg (HIGH) diet garcinol. After 14-day trial, three piglets per pen were chosen to collect plasma, intestinal tissue and colonic digesta samples. RESULTS: We demonstrated for the first time that garcinol promoted growth performance, as increased average daily feed intake (ADFI) and decreased feed/gain ratio (F/G); and reduced diarrhea incidence (P < 0.05); and strengthened antioxidant capacity, as an increased antioxidative index (P < 0.05). Additionally, garcinol ameliorated intestinal barrier dysfunction, as an increased villus height to crypt depth ratio, increased zonula occludens protein 1 (ZO-1), occludin and claudin-1 expression in the jejunum and ileum (P < 0.05), and decreased intestinal permeability (P < 0.05); and reduced inflammation, as decreased cytokine interleukin (IL)-6, IL-10, IL-1ß and tumor necrosis factor-α (TNF-α) levels in the mucosa of the jejunum and ileum, and NF-κB p65 translocation (P < 0.05). Moreover, garcinol inhibited the growth of most harmful bacteria in the gut, especially Escherichia coli, and increased the growth of the beneficial bacteria Lactobacillus. CONCLUSION: This work provides a fundamental basis for the future development of garcinol-functional food use for improving diarrhea and intestinal barrier function in weaned piglets and for understanding the biological effects of garcinol and its potential as a functional feed additive.

Dietary Garcinol Attenuates Hepatic Pyruvate and Triglyceride Accumulation by Inhibiting P300/CBP-Associated Factor in Mid-to-Late Pregnant Rats.

BACKGROUND: Increased hepatic glycolysis and lipogenesis are characteristic of pregnancy. OBJECTIVES: The present study aimed to investigate the mechanism of garcinol on the amelioration of hepatic pyruvate and triglyceride (TG) accumulation in mid-to-late pregnant rats. METHODS: Forty Sprague-Dawley pregnant rats (aged 9 wk, n = 10/diet) were fed a basal diet (control) or that diet plus garcinol at 100 ppm (Low Gar), 300 ppm (Mid Gar), or 500 ppm (High Gar) for 14 d. The livers were processed for Western blotting analyses and measuring enzymatic activity and pyruvate and TG concentrations. Hepatocytes from other pregnant Sprague Dawley rats were transfected with P300/CBP associating factor (PCAF) short interfering (si)RNAs; hepatocytes from nonpregnant Sprague-Dawley rats with overexpression of PCAF were treated with garcinol (5 µM). The activity and acetylation of upstream stimulatory factor (USF-1) and glycolytic enzymes were analyzed. RESULTS: Dietary garcinol significantly decreased (P < 0.05) concentrations of hepatic and plasma TG (27.1-45.8%) and total cholesterol (25.3-49.5%), plasma free fatty acids (24.4-37.8%), and hepatic pyruvate (31.5-43.5%) and lactate (33.4-65.7%) in mid-to-late pregnant rats. Garcinol promoted (P < 0.05) antioxidant capacity in the liver and plasma by 27.4-32.1%. Garcinol downregulated (P < 0.05) lipid synthesis-related enzyme expression by 30.6-85.3% and decreased (P < 0.05) glycolytic enzyme activities by 22.5-74.6% and PCAF activity by 18.6-55.4%. Transfection of PCAF siRNAs to hepatocytes of pregnant rats decreased USF-1 and glycolytic enzyme activities by PCAF; garcinol treatment downregulated (P < 0.05) the acetylation and activities of USF-1 and glycolytic enzymes by 35.6-83.7%. CONCLUSIONS: Garcinol attenuates hepatic pyruvate and TG accumulation in the liver of mid-to-late pregnant rats, which may be due to downregulating the acetylation of USF-1 and the glycolytic enzymes induced by PCAF in isolated hepatocytes.

The effect of dietary garcinol supplementation on oxidative stability, muscle postmortem glycolysis and meat quality in pigs.

The objective of this study was to evaluate the effects of dietary garcinol (0, 200, 400 and 600 mg/kg) on the growth performance, meat quality, postmortem glycolysis and antioxidative capacity of finishing pigs. Dietary garcinol increased pigs' average daily gain, pH 24h, a* and myoglobin content of longissimus dorsi (LM) (P < 0.05), and decreased feed/gain ratio, the L*24h, glycolytic potential, drip loss, shear force, and backfat depth (P < 0.05). The glutathione peroxidase (GPx), catalase (CAT) and total antioxidative capacity (T-AOC) were significantly increased by garcinol (P < 0.05), while the activity of lactate dehydrogenase (LDH) and malonaldehyde (MDA) content were decreased (P < 0.05). Moreover, garcinol decreased the p300/CBP-associated factor (PCAF) activity, the acetylation level and activities of glycolysis enzymes phosphoglycerate kinase 1 (PGK1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase-3 (PFKFB3) (P < 0.05). The results of this study showed that garcinol decreased postmortem glycolysis, and this may be due to the mechanism of decreasing glycolytic enzyme acetylation induced by PCAF. The present study indicates that garcinol can facilitate the growth performance of pigs and improve pork quality by changing postmortem glycolysis and antioxidative capacity.

Dietary supplementation with garcinol during late gestation and lactation facilitates acid-base balance and improves the performance of sows and newborn piglets1.

The present study was conducted to evaluate the effects of dietary garcinol supplementation during late gestation (from the 90th day of pregnancy; day 90) and lactation on the acid-base balance of the umbilical cord blood and performance of sows and piglets. Sixty sows (Duroc × Yorkshire × Landrace; second- or third-parity; n = 20) were randomly divided into 3 gestation (day 90 of pregnancy) or lactation treatments, control diet (CON; basal diet), basal diet with 200 mg garcinol, and basal diet with 600 mg garcinol per kg of feed. The body weight (BW); backfat thickness and litter size of the sows; and birth weight, weaning weight, and mortality of piglets were recorded. Sows' blood and piglets' umbilical cord blood were collected for the measurements of hematological parameters and antioxidative and immune indexes, and acid-base balance parameters, respectively. The colostrum and milk and fecal samples of the sows were also collected for analysis of milk composition and apparent total tract nutrient digestibility. Garcinol had no effect on the BW and backfat thickness of the sows but significantly increased the birth weight and weaning weight of piglets (P < 0.05) and decreased the mortality (P < 0.05). Moreover, the white blood cell counts and neutrophil count, mean cell hemoglobin, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) activity in the plasma of the sows were increased more significantly (P < 0.05) in the garcinol groups than that in the CON group, whereas the malondialdehyde (MDA) content was decreased (P < 0.05). The garcinol treatment significantly increased the pH, HCO3- and base excess values (P < 0.05), whereas it decreased the pCO2 and lactate content (P < 0.05) in the umbilical blood. Dry matter (DM), ash, and ether extract in the colostrum were similar between groups (P > 0.05), whereas the garcinol significantly increased the crude protein (CP) in the milk. In addition, the content of immunoglobulin A (IgA) and immunoglobulin G (IgG) in the plasma of piglets and in colostrum and milk of sows were increased more significantly (P < 0.05) in the garcinol groups than that in the CON group. The apparent total tract nutrient digestibility was similar between treatments. Collectively, this study indicates that sows fed with garcinol in late gestation and lactation showed improved maternal health and antioxidative status, milk protein content, acid-base balance in the umbilical cord blood, and growth performance in piglets, showing promise in natural plant extract nutrition for sows.

Enzymatic and nonenzymatic protein acetylations control glycolysis process in liver diseases.

Impaired glycolysis has pathologic effects on the occurrence and progression of liver diseases, and it appears that glycolysis is increased to different degrees in different liver diseases. As an important post-translational modification, reversible lysine acetylation regulates almost all cellular processes, including glycolysis. Lysine acetylation can occur enzymatically with acetyltransferases or nonenzymatically with acetyl-coenzyme A. Accompanied by the progression of liver diseases, there seems to be a temporal and spatial variation between enzymatic and nonenzymatic acetylations in the regulation of glycolysis. Here, we summarize the most recent findings on the functions and targets of acetylation in controlling glycolysis in the different stages of liver diseases. In addition, we discuss the differences and causes between enzymatic and nonenzymatic acetylations in regulating glycolysis throughout the progression of liver diseases. Then, we review these new discoveries to provide the potential implications of these findings for therapeutic interventions in liver diseases.-Li, J., Wang, T., Xia, J., Yao, W., Huang, F. Enzymatic and nonenzymatic protein acetylations control glycolysis process in liver diseases.

The Variation of Nasal Microbiota Caused by Low Levels of Gaseous Ammonia Exposure in Growing Pigs.

Exposure to gaseous ammonia, even at low levels, can be harmful to pigs and human health. However, less is known about the effects of sustained exposure to gaseous ammonia on nasal microbiota colonization in growing pigs. A total of 120 Duroc×Landrace×Yorkshire pigs were housed in 24 separate chambers and continuously exposed to gaseous ammonia at 0,5, 10, 15, 20, and 25 ppm (four groups per exposure level) for 4 weeks. Then, we used high-throughput sequencing to perform 16S rRNA gene analysis in nasal swabs samples from 72 pigs (n = 12). The results of the nasal microbiota analysis showed that an increase in ammonia concentration, especially at 20 and 25 ppm, decreased the alpha diversity and relative abundance of nasal microbiota. Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, and Chloroflexi were the most abundant phyla. In addition, the relative abundances of 24 microbial genera significantly changed as the ammonia level increased. Four microbial genera (Pseudomonas, Lactobacillus, Prevotella, and Bacteroides) were significantly decreased at 25 ppm, while only two genera (Moraxella and Streptococcus) were increased at 25 ppm. PICRUSt analyses showed that the relative abundances of the nasal microbiota involved in cell motility, signal transduction, the nervous system, environmental adaptation, and energy and carbohydrate metabolism were significantly decreased, while genes involved in the immune system, endocrine system, circulatory system, immune system diseases and metabolism of vitamins, lipid, and amino acids were increased with increased ammonia levels. The results of in vivo tests showed that an increase in ammonia levels, especially an ammonia level of 25 ppm, caused respiratory tract injury and increase the number of Moraxella and Streptococcus species, while simultaneously decreasing respiratory immunity and growth performance, consistent with the increased presence of harmful bacteria identified by nasal microbiota analysis. Herein, this study also indicted that the threshold concentration of ammonia in pig farming is 20 ppm.

Glucagon-Induced Acetylation of Energy-Sensing Factors in Control of Hepatic Metabolism.

The liver is the central organ of glycolipid metabolism, which regulates the metabolism of lipids and glucose to maintain energy homeostasis upon alterations of physiological conditions. Researchers formerly focused on the phosphorylation of glucagon in controlling liver metabolism. Noteworthily, emerging evidence has shown glucagon could additionally induce acetylation to control hepatic metabolism in response to different physiological states. Through inducing acetylation of complex metabolic networks, glucagon interacts extensively with various energy-sensing factors in shifting from glucose metabolism to lipid metabolism during prolonged fasting. In addition, glucagon-induced acetylation of different energy-sensing factors is involved in the advancement of nonalcoholic fatty liver disease (NAFLD) to liver cancer. Here, we summarize the latest findings on glucagon to control hepatic metabolism by inducing acetylation of energy-sensing factors. Finally, we summarize and discuss the potential impact of glucagon on the treatment of liver diseases.