Tauroursodeoxycholic acid(TUDCA)improves intestinal barrier function associated with TGR5-MLCK pathway and the alteration of serum metabolites and gut bacteria in weaned piglets

Background:Tauroursodeoxycholic acid(TUDCA),a hydrophilic bile acid,is the main medicinal component of bear bile and is commonly used to treat a variety of hepatobiliary diseases.Meanwhile,TUDCA has been shown to modulate the intestinal barrier function and alleviate DSS-induced colitis in mice.However,the effect of TUDCA on the intestinal barrier of weaned piglets remains largely unclear.Methods:The weaned piglets and porcine IPEC-J2 intestinal epithelial cells were used to investigate the effects of TUDCA on intestinal barrier function in weaned piglets and explore the possible underlying mechanisms.In vivo,72 healthy weaned piglets were randomly allocated into 2 groups according to their gender and body weight,and piglets were fed the basal diet with 0(control,CON)and 200 mg/kg TUDCA for 30 d,respectively.Three female and three male piglets reflecting the average bodyweight were slaughtered in each group and samples were collected.In vitro,IPEC-J2 cells were subjected to 100μmol/L TUDCA to explore the possible underlying mechanisms.Results:Our results demonstrated that dietary TUDCA supplementation significantly reduced the diarrhea incidence of weaned piglets,possibly attributing to the TUDCA-enhanced intestinal barrier function and immunity.In addition,TUDCA supplementation altered serum metabolites and the relative abundance of certain gut bacteria,which might contribute to the improved intestinal barrier function.Furthermore,the in-vitro results showed that TUDCA improved the E.coli-induced epithelial barrier impairment of IPEC-J2 cells and increased Takeda G-coupled protein receptor 5(TGR5)protein expression.However,knockdown of TGR5 and inhibition of myosin light chain kinase(MLCK)pathway abolished the TUDCA-improved epithelial barrier impairment in E.coli-treated IPEC-J2 cells,indicating the involvement of TGR5-MLCK in this process.Conclusions:These findings showed that TUDCA improved intestinal barrier function associated with TGR5-MLCK pathway and the alteration of serum metabolites and gut bacteria in weaned piglets,suggesting the potential application of TUDCA in improving gut health in piglet production.

Effects of Probiotics on Growth, Pork Quality and Serum Metabolites in Growing-finishing Pigs

A total of 72 growing-finishing pigs（56 days old） were used to investigate the effects of dietary probiotics on growing performance, pork quality and serum metabolites. Using single factorial experiment design, pigs were allotted to three groups（four pens/group and six pigs/pen）. Diet treatments were a corn-soybean meal（control group） and a corn-soybean meal with 1% probiotics（probiotics group） or 80 mg kg-1colistin sulfate（antibiotics group）. Pigs were slaughtered when pigs of the lightest averaged 90 kg（157 days of age）. There was no difference（p＆gt;0.05） in average daily gain or feed/gain, and the average daily feed intake of the antibiotics group was higher than that of the probiotics and the control groups（p＆lt;0.05）. Diet supplemented with probiotics reduced the drip loss and cooking loss of pork（p＆lt;0.05）. Dietary supplementation with probiotics significantly decreased the content of triglycerides, lowdensity lipoprotein and urea nitrogen in serum（p＆lt;0.05）, and increased the content of high-density lipoprotein in serum（p＆lt;0.05）. These results suggested improving effects of dietary probiotics on pork quality and serum metabolism in growing-finishing pigs.

A comparison of average daily gain,apparent digestibilities,energy balance,rumen fermentation parameters,and serum metabolites between yaks(Bos grunniens)and Qaidam cattle(Bos taurus)consuming diets differing in energy level

Yaks(Bos grunniens),indigenous to the harsh Qinghai-Tibetan Plateau,are well adapted to the severe conditions,and graze natural pasture without supplements all year round.Qaidam cattle(Bos taurus),introduced to the Qinghai-Tibetan Plateau 1,700 years ago,are raised at a lower altitude than yaks,provided with shelter at night and offered supplements in winter.Based on their different backgrounds,we hypothesized that yaks have lower energy requirements for maintenance than cattle.To test this hypothesis,we measured average daily gain(ADG),apparent digestibilities,energy balance,rumen fermentation parameters,and serum metabolites in growing yaks and cattle offered diets differing in metabolizable energy(ME)levels(6.62,8.02,9.42 and 10.80 MJ/kg),but with the same crude protein concentration.Six castrated yaks(155±5.8 kg)and 6 castrated Qaidam cattle(154±8.0 kg),all 2.5 years old,were used in 2 concurrent 4×4 Latin square designs.Neutral and acid detergent fiber digestibilities were greater(P<0.05)in yaks than in cattle,and decreased linearly(P<0.05)with increasing dietary energy level;whereas,digestibilities of dry matter,organic matter,crude protein and ether extract increased(P<0.05)linearly with increasing energy level.The ADG was greater(P<0.001)in yaks than in cattle,and increased(P<0.05)linearly with increasing energy levels.From the regressions of ADG on ME intake,the estimated ME requirement for maintenance was lower(P<0.05)in yaks than in cattle(0.43 vs.0.57 MJ/kg BW0.75).The ratios of digestible energy(DE):gross energy and ME:DE were higher(P<0.05)in yaks than in cattle,and increased(P<0.05)linearly with increasing dietary energy level.Ruminal pH decreased(P<0.05),whereas concentrations of total volatile fatty acids(VFAs)and ammonia increased(P<0.01)with increasing dietary energy level,and all were greater(P<0.05)in yaks than in cattle.Concentrations of ruminal acetate and iso-VFAs were greater(P<0.05),whereas propionate was lower(P<0.05)in yaks than in cattle;acetate decreased(P<0.001),whereas butyrate and propionate increased(P<0.001)linearly with increasing dietary energy level.Serum concentrations ofβ-hydroxybutyrate were lower(interaction,P<0.001)in yaks than in cattle fed diets of 9.42 and 10.80 MJ/kg,whereas non-esterified fatty acids were greater(interaction,P<0.01)in yaks than in cattle fed diets of 6.62 and 8.02 MJ/kg.Concentrations of serum leptin and growth hormone were greater in yaks than in cattle and serum insulin and growth hormone increased(P<0.01)linearly with increasing dietary energy level.Our hypothesis that yaks have lower energy requirements for maintenance than cattle was supported.This lower requirement confers an advantage to yaks over Qaidam cattle in consuming low energy diets during the long winter on the Qinghai-Tibetan Plateau.

Effects of dietary supplementation with Allium mongolicum Regel extracts on growth performance,serum metabolites,immune responses,antioxidant status,and meat quality of lambs

The present study was to evaluate the effects of dried Allium mongolicum Regel(AMR)powder and its water-and fat-soluble extracts(AWE and AFE)on the growth performance,serum metabolites,immune responses,antioxidant status,and meat quality of lambs.A total of 32 male small-tailed Han lambs(5 months old;initial body weight=34.8±0.40 kg)were used in a 60-d feeding experiment after a 15-d adaptation period.The lambs were randomly divided into 4 groups(n=8)and fed a basal diet(control,CON group),the basal diet supplemented with dried AMR powder at 10 g/d per lamb(AMR group),the basal diet supplemented with AWE at 3.4 g/d per lamb(AWE group),or the basal diet supplemented with AFE at 2.8 g/d per lamb(AFE group).Blood samples were collected on d 0,30,and 60 in the feeding experiment(n-8).At the end of the experiment,the lambs were sacrificed and the longissimus dorsi muscles collected.Growth performance was not significantly affected by dietary supplementation of AMR,AWE and AFE(P>0.05).However,significantly lower albumin(P=0.006),total protein(P=0.006),globin(P=0.025),and blood urea nitrogen(P=0.024)concentrations were observed in AFE group relative to CON and AMR groups.Similarly,a significantly lower lactate dehydrogenase activity(P=0.018)was observed in AFE group relative to AWE group,but not in other groups(P>0.05).In addition,significantly increasing trends in glutathione peroxidase(P=0.06)in AMR,AWE,and AFE groups were observed relative to the control group.Furthermore,significantly lower drip loss(P-0.011)across the treatment groups and cooking loss(P-0.048)were observed in the AMR group relative to the control group.Taken together,these results indicate that AMR and its extracts had no significant effect on lamb growth performance,antioxidant status,and immune responses,but could significantly improve meat quality without the occurrence of pathological kidney and liver lesions.

Magnesium-L-threonate treats Alzheimer's disease by modulating the microbiota-gut-brain axis

Disturbances in the microbiota-gut-brain axis may contribute to the development of Alzheimer's disease. Magnesium-L-threonate has recently been found to have protective effects on learning and memory in aged and Alzheimer's disease model mice. However, the effects of magnesium-L-threonate on the gut microbiota in Alzheimer's disease remain unknown. Previously, we reported that magnesium-L-threonate treatment improved cognition and reduced oxidative stress and inflammation in a double-transgenic line of Alzheimer's disease model mice expressing the amyloid-β precursor protein and mutant human presenilin 1(APP/PS1). Here, we performed 16S r RNA amplicon sequencing and liquid chromatography-mass spectrometry to analyze changes in the microbiome and serum metabolome following magnesium-Lthreonate exposure in a similar mouse model. Magnesium-L-threonate modulated the abundance of three genera in the gut microbiota, decreasing Allobaculum and increasing Bifidobacterium and Turicibacter. We also found that differential metabolites in the magnesiumL-threonate-regulated serum were enriched in various pathways associated with neurodegenerative diseases. The western blotting detection on intestinal tight junction proteins(zona occludens 1, occludin, and claudin-5) showed that magnesium-L-threonate repaired the intestinal barrier dysfunction of APP/PS1 mice. These findings suggest that magnesium-L-threonate may reduce the clinical manifestations of Alzheimer's disease through the microbiota-gut-brain axis in model mice, providing an experimental basis for the clinical treatment of Alzheimer's disease.

Treadmill exercise in combination with acousto-optic and olfactory stimulation improves cognitive function in APP/PS1 mice through the brain-derived neurotrophic factor-and Cygb-associated signaling pathways

A reduction in adult neurogenesis is associated with behavioral abnormalities in patients with Alzheimer's disease.Consequently,enhancing adult neurogenesis represents a promising therapeutic approach for mitigating disease symptoms and progression.Nonetheless,nonpharmacological interventions aimed at inducing adult neurogenesis are currently limited.Although individual non-pharmacological interventions,such as aerobic exercise,acousto-optic stimulation,and olfactory stimulation,have shown limited capacity to improve neurogenesis and cognitive function in patients with Alzheimer's disease,the therapeutic effect of a strategy that combines these interventions has not been fully explored.In this study,we observed an age-dependent decrease in adult neurogenesis and a concurrent increase in amyloid-beta accumulation in the hippocampus of amyloid precursor protein/presenilin 1 mice aged 2-8 months.Amyloid deposition became evident at 4 months,while neurogenesis declined by 6 months,further deteriorating as the disease progressed.However,following a 4-week multifactor stimulation protocol,which encompassed treadmill running(46 min/d,10 m/min,6 days per week),40 Hz acousto-optic stimulation(1 hour/day,6 days/week),and olfactory stimulation(1 hour/day,6 days/week),we found a significant increase in the number of newborn cells(5'-bromo-2'-deoxyuridine-positive cells),immature neurons(doublecortin-positive cells),newborn immature neurons(5'-bromo-2'-deoxyuridine-positive/doublecortin-positive cells),and newborn astrocytes(5'-bromo-2'-deoxyuridine-positive/glial fibrillary acidic protein-positive cells).Additionally,the amyloid-beta load in the hippocampus decreased.These findings suggest that multifactor stimulation can enhance adult hippocampal neurogenesis and mitigate amyloid-beta neuropathology in amyloid precursor protein/presenilin 1 mice.Furthermore,cognitive abilities were improved,and depressive symptoms were alleviated in amyloid precursor protein/presenilin 1 mice following multifactor stimulation,as evidenced by Morris water maze,novel object recognition,forced swimming test,and tail suspension test results.Notably,the efficacy of multifactor stimulation in consolidating immature neurons persisted for at least 2weeks after treatment cessation.At the molecular level,multifactor stimulation upregulated the expression of neuron-related proteins(NeuN,doublecortin,postsynaptic density protein-95,and synaptophysin),anti-apoptosis-related proteins(Bcl-2 and PARP),and an autophagyassociated protein(LC3B),while decreasing the expression of apoptosis-related proteins(BAX and caspase-9),in the hippocampus of amyloid precursor protein/presenilin 1 mice.These observations might be attributable to both the brain-derived neurotrophic factor-mediated signaling pathway and antioxidant pathways.Furthermore,serum metabolomics analysis indicated that multifactor stimulation regulated differentially expressed metabolites associated with cell apoptosis,oxidative damage,and cognition.Collectively,these findings suggest that multifactor stimulation is a novel non-invasive approach for the prevention and treatment of Alzheimer's disease.

Hydrogen-rich water alleviates constipation by attenuating oxidative stress through the sirtuin1/nuclear factor-erythroid-2-related factor 2/heme oxygenase-1 signaling pathway

BACKGROUND Constipation,a highly prevalent functional gastrointestinal disorder,induces a significant burden on the quality of patients'life and is associated with substantial healthcare expenditures.Therefore,identifying efficient therapeutic modalities for constipation is of paramount importance.Oxidative stress is a pivotal contributor to colonic dysmotility and is the underlying pathology responsible for constipation symptoms.Consequently,we postulate that hydrogen therapy,an emerging and promising intervention,can serve as a safe and efficacious treatment for constipation.AIM To determine whether hydrogen-rich water(HRW)alleviates constipation and its potential mechanism.METHODS Constipation models were established by orally loperamide to Sprague-Dawley rats.Rats freely consumed HRW,and were recorded their 24 h total stool weight,fecal water content,and charcoal propulsion rate.Fecal samples were subjected to 16S rDNA gene sequencing.Serum non-targeted metabolomic analysis,malondialdehyde,and superoxide dismutase levels were determined.Colonic tissues were stained with hematoxylin and eosin,Alcian blue-periodic acid-Schiff,reactive oxygen species(ROS)immunofluorescence,and immunohistochemistry for cell growth factor receptor kit(c-kit),PGP 9.5,sirtuin1(SIRT1),nuclear factor-erythroid-2-related factor 2(Nrf2),and heme oxygenase-1(HO-1).Quantitative real-time PCR and western blot analysis were conducted to determine the expression level of SIRT1,Nrf2 and HO-1.A rescue experiment was conducted by intraperitoneally injecting the SIRT1 inhibitor,EX527,into constipated rats.NCM460 cells were induced with H2O2 and treated with the metabolites to evaluate ROS and SIRT1 expression.RESULTS HRW alleviated constipation symptoms by improving the total amount of stool over 24 h,fecal water content,charcoal propulsion rate,thickness of the intestinal mucus layer,c-kit expression,and the number of intestinal neurons.HRW modulated intestinal microbiota imbalance and abnormalities in serum metabolism.HRW could also reduce intestinal oxidative stress through the SIRT1/Nrf2/HO-1 signaling pathway.This regulatory effect on oxidative stress was confirmed via an intraperitoneal injection of a SIRT1 inhibitor to constipated rats.The serum metabolites,β-leucine(β-Leu)and traumatic acid,were also found to attenuate H2O2-induced oxidative stress in NCM460 cells by up-regulating SIRT1.CONCLUSION HRW attenuates constipation-associated intestinal oxidative stress via SIRT1/Nrf2/HO-1 signaling pathway,modulating gut microbiota and serum metabolites.β-Leu and traumatic acid are potential metabolites that upregulate SIRT1 expression and reduce oxidative stress.

Integrated network pharmacology and metabolomics to explore the mechanisms of Shenzao dripping pill against chronic myocardial ischemia

Background:Shenzao dripping pill(SZDP)is empirically prescribed for treating cardiac diseases.Nevertheless,there is a lack of comprehensive knowledge regarding the underlying mechanisms contributing to its therapeutic effects.The objective of this study is to investigate the underlying mechanism of SZDP against chronic myocardial ischemia(CMI)in a rat model.Methods:In this study,we utilized electrocardiographic and echocardiographic detection along with pathological tissue analysis to evaluate the efficacy of SZDP.The integration of network pharmacology and metabolomics was conducted to investigate the mechanisms.Molecular docking and molecular dynamics simulations were used to validate the binding energy between the compounds of SZDP and the associated targets.Results:The results showed that SZDP was able to improve T wave voltage,reverse CMI abnormalities in ejection fraction and fractional shortening,and restore histopathological heart damage.Metabolomics results indicated that disturbances of metabolic profile in CMI rats were partly corrected after SZDP administration,mainly affecting purine metabolism.13-Docosenamide may be the potential metabolic biomarker of the therapeutic application of SZDP for CMI.Integrating network pharmacology and metabolomics,thiopurine S-methyltransferase(TPMT),xanthine dehydrogenase/oxidase(XDH),bifunctional purine biosynthesis protein ATIC(ATIC),and cytochrome p4501A1(CYP1A1)were identified as possible targets of SZDP to exert therapeutic effects by enhancing the metabolic levels of L-Tryptophan,Deoxyribose 1-phosphate and Phosphoribosyl formamidocarboxamide.Conclusion:SZDP has a therapeutic effect on CMI by regulating metabolite levels,acting on the targets of TMPT,XDH,ATIC,and CYP1A1,and reducing cardiomyocyte injury and myocardial fibrosis.

Bile acids as drivers and biomarkers of hepatocellular carcinoma

The prevalence of hepatocellular carcinoma(HCC)is rapidly increasing,driven not least in part by the escalating prevalence of non-alcoholic fatty liver disease.Bile acid(BA)profiles are altered in patients with HCC and there is a developing body of evidence from in vitro human cellular models as well as rodent data suggesting that BA are able to modulate fundamental processes that impact on cellular phenotype predisposing to the development of HCC including senescence,proliferation and epithelial-mesenchymal transition.Changes in BA profiles associated with HCC have the potential to be exploited clinically.Whilst excellent diagnostic and imaging tools are available,their use to screen populations with advanced liver disease at risk of HCC is limited by high cost and low availability.The mainstay for HCC screening among subjects with cirrhosis remains frequent interval ultrasound scanning.Importantly,currently available serum biomarkers add little to diagnostic accuracy.Here,we review the current literature on the use of BA measurements as predictors of HCC incidence in addition to their use as a potential screening method for the early detection of HCC.Whilst these approaches do show early promise,there are limitations including the relatively small cohort sizes,the lack of a standardized approach to BA measurement,and the use of inappropriate control comparator samples.

Occurrence of retained placenta is preceded by an inflammatory state and alterations of energy metabolism in transition dairy cows

Background： Failure to expel fetal membranes within 24 h of calving is a pathological condition defined as retained placenta（RP）. The objective of this investigation was to evaluate whether there are alterations in several selected serum variables related to innate immunity and carbohydrate and lipid metabolism that precede occurrence of RP in transition Holstein dairy cows.Methods： One hundred multiparous Holstein dairy cows were involved in the study. Blood samples were collected from the coccygeal vein during the-8 to ＋4 wks around parturition, once per week before the morning feeding.Six healthy control cows（CON） and 6 cows with RP were selected and serum samples at-8,-4, time of diagnosis of disease, and ＋4 wks relative to parturition were used for analyses. All samples were analyzed for lactate, nonesterified fatty acids（NEFA）, β-hydroxybutyrate（BHBA）, interleukin-1（IL-1）, interleukin-6（IL-6）, tumor necrosis factor（TNF）, haptoglobin（Hp）, and serum amyloid A（SAA）.Results： Cows with RP had greater concentrations of serum lactate, IL-1, IL-6, TNF, and SAA in comparison with CON cows. Intriguingly, elevated concentrations of all five variables were observed at-8 and-4 wks before the occurrence of RP compared to healthy cows. Cows with RP also had lower DMI and milk production vs CON animals; however milk composition was not affected by RP.Conclusions： Cows with RP showed an activated innate immunity 8 wks prior to diagnosis of disease. Overall results suggest that serum IL-1, IL- 6, and TNF, and lactate can be used as screening biomarkers to indicate cows that might have health issues during the transition period.

Spraying compound probiotics improves growth performance and immunity and modulates gut microbiota and blood metabolites of suckling piglets

One factor that shapes the establishment of early neonatal intestinal microbiota is environmental microbial exposure,and probiotic application has been shown to promote health and growth of piglets.Thus,this study hypothesized that environmental probiotic application in early days of life would be beneficial to newborn piglets.This study aimed to investigate the effect of spraying a compound probiotic fermented liquid(CPFL)into the living environment of piglets on their early growth performance and immunity.This work included 68 piglets,which were randomized into probiotic and control groups.Blood and fecal samples were collected at 0,3,7,14,and 21 days of age.Spraying CPFL significantly reshaped the microbiota composition of the delivery room environment,increased piglets’daily weight gain and weaning weight(P<0.001),and modulated piglets’serum cytokine levels(increases in Ig A,Ig G,and IL-10;decrease in IFN-γ;P<0.05 in each case)in piglets.Additionally,spraying CPFL during early days of life modified piglets’gut microbiota structure and diversity,increased the abundance of some potentially beneficial bacteria(such as Bacteroides uniformis,Butyricimonas virosa,Parabacteroides distasonis,and Phascolarctobacterium succinatutens)and decreased the abundance of Escherichia coli(P<0.05).Interestingly,CPFL application also significantly enhanced the gut microbial bioactive potential and levels of several serum metabolites involved in the metabolism of vitamins(B2,B3,B6,and E),medium/long-chain fatty acids(caproic,tetradecanoic,and peptadecanoic acids),and dicarboxylic acids(azelaic and sebacic acids).Our study demonstrated that spraying CPFL significantly could improve piglets’growth performance and immunity,and the beneficial effects are associated with changes in the gut microbiota and host metabolism.Our study has provided novel data for future development of probiotic-based health-promoting strategies and expanded our knowledge of probiotic application in animal husbandry.

Dietary chenodeoxycholic acid improves growth performance and intestinal health by altering serum metabolic profiles and gut bacteria in weaned piglets

Nutritional diarrhea and subsequent performance degradation in weaned piglets are major challenges for the pig industry.Bile acids(BA)can be added to the diet as emulsifiers.This experiment was conducted to investigate the effects of chenodeoxycholic acid(CDCA),a major primary BA,on growth performance,serum metabolic profiles and gut health in weaned piglets.A total of 72 healthy weaned piglets were randomly assigned to the control(CON)and the CDCA groups,which were feed a basal diet and the basal diet supplemented with 200 mg/kg CDCA for 30 d,respectively.Our results demonstrated that CDCA significantly increased final BW and average daily gain(ADG),decreased feed-to-gain(F:G)ratio and tended to reduce diarrhea incidence.In addition,CDCA increased the villus height-to-crypt depth(V:C)ratio,elevated goblet cell numbers and the expression of tight junction proteins,suggesting the enhancement of intestinal barrier function.As an emulsifier,CDCA increased jejunal lipase activity and the mRNA expression of pancreatic lipases.CDCA supplementation also altered the serum metabolic profiles,including increasing the levels of indole 3-acetic acid,N'-formylkynurenine and theobromine that were beneficial for gut health.Moreover,the relative abundance of 2 beneficial gut bacteria,Pre-votella 9 and Prevotellaceae TCG-001,were increased,whereas the relative abundance of a harmful bacteria,Dorea,was decreased in the gut of weaned piglets supplemented with CDCA.Importantly,the altered serum metabolic profiles showed a strong correlation with the changed gut bacteria.In conclusion,CDCA improved the growth performance of weaned piglets by improving intestinal morphology and barrier function,and enhancing lipid digestion,accompanied by alterations of serum metabolic profiles,and changes in relative abundance of certain gut bacteria.

Growth performance,feed utilization,body and fatty acid composition of Nile tilapia (Oreochromis niloticus) fed diets containing elevated levels of palm oil

This study was conducted to evaluate effects of dietary palm oil levels on growth performance,feed utilization,body composition and serum metabolites of Oreochromis niloticus.Five isonitrogenous diets,32% crude protein with increasing palm oil levels of 0(Control,CTRL),2%,4%,6% and 8% were used as the major lipid source for the trial.The results showed that greatest weight gain,specific growth rate,and protein efficiency ratio occurred at 6% dietary palm oil level.Dietary palm oil levels significantly(P<0.05)affected lipid,moisture,ash and crude protein contents in muscle and whole body.Serum triglycerides,cholesterol and total protein were significantly affected by elevated palm oil levels.Furthermore,total saturates,total n-3 poly unsaturated fatty acids(PUFA),total n-6 PUFA as well as DHA/EPA in muscle were significantly affected by different levels of palm oil.Fish fed the 6% palm oil level recorded the highest level of whole body docosahexaenoic acid(DHA),which was significantly higher than that of the control.The nutritional quality of O.niloticus was altered by different palm oil levels.The present study suggests that 6% dietary palm oil is the best feed formulation for tilapia,Oreochromis niloticus.