Interaction of Soybean Phospholipids Based Lipo-Vesicle and Surfactants of Bile Salts In Vitro

Liposomes were prepared with natural soybean phospholipids by extrusion method after rotating-film evaporating technique. Transmission electron micrography was used to detect the appearances of the prepared liposomes, and the liposome diameter was also measured. The prepared liposomes were sphere in shape with the mean diameter of 217 nm and span of 0.838. The phospholipid bilayer structure, suitable for entrapping various effector molecules, could be seen clearly under transmission electron microscopy. The bile salts of sodium cholate and sodium deoxycholate were used as the surfactants to investigate their interaction with liposomes. The turbidities for the mixture of bile salts and liposomes were evaluated by the visible spectrometry method at the wavelength of 500 nm. And the diameter changes of liposomes were also tested to examine the effect of bile salts on liposomes. At the beginning, the diameters and turbidities of liposomes increased a little as the result of mixed micelles formation during the different stages for the structure changes of surfactant-liposomes micelles. The further added bile salts decreased the diameters and turbidities of liposomes. The liposome suspension underwent several rearrangements before small mixed micelles formed. And the diameter of liposomes changed regularly. The interaction of bile salts and liposomes is important for the further study of the behaviors of liposomes in vivo. The drug loaded and release properties of liposomes can also be well reflected by the interaction of liposomes and surfactants.

Increased susceptibility for intrahepatic cholestasis of pregnancy and contraceptive-induced cholestasis in carriers of the 1331T>C polymorphism in the bile salt export pump

AIM： To study the association of three common ABCB11 and ABCC2 polymorphisms （ABCB11： 1331T〉C→V444A; ABCC2： 3563T〉A → V1188E and 4544G 〉A → C1515Y） with intrahepatic cholestasis of pregnancy （ICP） and contraceptive-induced cholestasis （CIC）. METHODS： ABCB11 and ABCC2 genotyping data were available from four CIC patients and from 42 and 33 ICP patients, respectively. Allele-frequencies of the studied polymorphisms were compared with those in healthy pregnant controls and Caucasian individuals. Furthermore, serum bile acid levels were correlated with the presence or absence of the 1331 C allele. RESULTS： The ABCB11 1331T〉C polymorphism was significantly more frequent in cholestatic patients than in pregnant controls： C allele 76.2% （CI, 58.0-94.4） vs 51.3% （CI 35.8-66.7）, respectively （P = 0.0007）; and CC allele 57.1% （CI 36.0-78.3） vs 20% （CI 7.6-32.4）, respectively （P = 0.0065）. All four CIC patients were homozygous carriers of the C allele. In contrast, none of the studied ABCC2 polymorphism was overrepresented in ICP or CIC patients. Higher serum bile acid levels were found in carriers of the 1331CC genotype compared to carriers of the TT genotype. CONCLUSION： Our data support a role for the ABCB11 1331T〉C polymorphism as a susceptibility factor for the development of estrogen-induced cholestasis, whereas no such association was found for ABCC2. Serum bile acid and 7-glutamyl transferase levels might help to distinguish ABCB4- and ABCB11-related forms of ICP and CIC.

Bile salts inhibit growth and induce apoptosis of culture human normal esophageal mucosal epithelial cells

AIM： To investigate the effect of six bile salts： glycocholate （GC）, glycochenodeoxycholate （GCDC）, glycodeoxycholate （GDC）, taurocholate （TC）, taurochenodeoxycholate （TCDC）, taurodeoxycholate （TDC）, and their mixture on cultured human normal esophageal rnucosal epithelial cells. METHODS： Human normal esophageal mucosal epithelial cells were cultured with serum-free keratinocyte medium. 3-[4,5-Dimethylthiaolyl]-2,5- diphenyl-tetrazolium bromide assay was applied to the detection of cell proliferation. Apoptotic morphology was observed by phase-contrast video microscopy and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） assay. Sub-G1 DNA fragmentations and early apoptotic cells were assayed by flow cytometry （FCI） with propidium iodide （PI） staining and annexin V-FITC conjugated with PI staining. Apoptotic DNA ladders on agarose gel electrophoresis were observed. RESULTS： Except for GC, GCDC, GDC, TC, TCDC, TDC and their mixture could initiate growth inhibition of esophageal mucosal epithelial cells in a dose- and time-dependent manner. TUNEL and FCM assays demonstrated that the bile salts at 500 μmol/L and their mixture at 1 500 μmol/L induced apoptosis except for GC. The percentage of sub-G1 detected by FCM with PI staining was 83.5% in cells treated with 500 μmol/L TC for 2 h, and 19.8%, 20.4%, 25.6%, 13.5%, and 75.8% in cells treated with 500 μmol/L GCDC, TCDC, GDC, TDC, and 1 500 μmol/L mixture for 24 h, respectively, which were higher than that of the control （1.5%）. The percentage was 1.4% in cells with 500 μmol/L GC for 24 h. DNA ladders on agarose gel electrophoresis were seen in cells treated with 500 μmol/L TC for 2 h and i 500 μmnol/L mixture for 24 h. CONCLUSION： All GCDC, GDC, TC, TCDC, TDC and their mixture can inhibit growth and induce apoptosis of cultured human normal esophageal mucosal epithelial cells, but GC is well tolerated by the cells.

Bile salts inhibit growth and induce apoptosis of human esophageal cancer cell line

AIM： To explore the effect of six bile salts, including glycocholate （GC）, glycochenodeoxycholate （GCEX：）, glycodeoxycholate （GDC）, taurocholate （TC）, taurochenodeoxycholate （TCDC）, taurodeoxycholate （TDC）, and two bile acids including cholic acid （CA） and deoxycholic acid （DCA） on esophageal cancer Eca109 cell line. METHODS： Eca109 cells were exposed to six bile salts, two bile adds and the mixed bile salts at different concentrations for 24-72 h. 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide （MTT） assay was used to detect the cell proliferation. Apoptotic morphology was observed by phase-contrast video microscopy and deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） assay. Sub-G1 DNA fragmentations and early apoptosis cells were assayed by flow cytometry （FCM） with propidium iodide （PI） staining and annexin V-FITC conjugated with PI staining. Apoptosis DNA ladders on agarose were observed. Activation of caspase-3 was assayed by FCM with FITC-conjugated monodonal rabbit anti-active caspase- 3 antibody and expressions of Bcl-2 and Bax proteins were examined immunocytochemically in 500μmol/L-TCinduced apoptosis cells. RESULTS： Five bile salts except for GC, and two bile acids and the mixed bile salts could initiate growth inhibition of Ecal09 cells in a dose- and time-dependent manner. TUNEL, FCM, and DNA ladder assays all demonstrated apoptosis induced by bile salts and bile acids at 500 μmol/L, except for GC. Early apoptosis cell percentages in Eca109 cells treated with GCDC, GDC, TC, TCDC, TDC, CA at 500 μmol/L for 12 h, DCA at 500 μmol/L for 6 h, and mixed bile salts at 1 000 μmol/L for 12 h were 7.5%, 8.7%, 14.8%, 8.9%, 7.8%, 9.3%, 22.6% and 12.5%, respectively, all were significantly higher than that in control （1.9%）. About 22% of the cell population treated with TC at 500 μmol/L for 24 h had detectable active caspase-3, and were higher than that in the control （1%）. Immunocytochemical assay suggested that TC down-regulated Bcl-2 protein level and up-regulated Bax protein level. CONCLUSION： GCDC, GDC, TC, TCDC, TDC, CA and DCA, except for GC, can inhibit growth and induce apoptosis of esophageal cancer Eca109 cells. Activation of caspase-3, decreased Bcl-2 protein and increased Bax protein are involved in TC-induced apoptosis of Eca109 cells.

Partial external biliary diversion in bile salt export pump deficiency: Association between outcome and mutation

To investigate the relation of two different mutations to the outcome of partial external biliary diversion (PEBD) in severe bile salt export pump (BSEP) deficiency. METHODSMutations in the gene encoding BSEP leading to severe BSEP deficiency in two unrelated patients were identified by genomic sequencing. Native liver biopsies and transiently transfected human embryonic kidney (HEK) 293 cells expressing either wild-type or mutated BSEP were subjected to immunofluorescence analysis to assess BSEP transporter localization. Bile acid profiles of patient and control bile samples were generated by ultra-performance liquid chromatography-tandem mass spectrometry. Wild-type and mutant BSEP transport of [3H]-labeled taurocholate (TC) and taurochenodeoxycholate (TCDC) was assessed by vesicular transport assays. RESULTSA girl (at 2 mo) presented with pruritus, jaundice and elevated serum bile salts (BS). PEBD stabilized liver function and prevented liver transplantation. She was heterozygous for the BSEP deletion p.T919del and the nonsense mutation p.R1235X. At the age of 17 years relative amounts of conjugated BS in her bile were normal, while total BS were less than 3% as compared to controls. An unrelated boy (age 1.5 years) presenting with severe pruritus and elevated serum BS was heterozygous for the same nonsense and another missense mutation, p.G1032R. PEBD failed to alleviate pruritus, eventually necessitating liver transplantation. BS concentration in bile was about 5% of controls. BS were mainly unconjugated with an unusual low amount of chenodeoxycholate derivatives (< 5%). The patients’ native liver biopsies showed canalicular BSEP expression. Both BSEP p.T919del and p.G1032R were localized in the plasma membrane in HEK293 cells. In vitro transport assays showed drastic reduction of transport by both mutations. Using purified recombinant BSEP as quantifiable reference, per-molecule transport rates for TC and TCDC were determined to be 3 and 2 BS molecules per wild-type BSEP transporter per minute, respectively. CONCLUSIONIn summary, our findings suggest that residual function of BSEP as well as substrate specificity influence the therapeutic effectiveness of PEBD in progressive familial intrahepatic cholestasis type 2 (PFIC-2).

Cloning of Bile Salt Hydrolase Gene and Its Expression in Lactic Acid Bacteria

According to the sequence of the bile salt hydrolase （BSH） gene of Bifidobacterium and the restriction enzyme cutting sites of expression vector pNZ8148, primers were designed and the bile salt hydrolase （BSH） gene was gotten from Bacillus bifidus ATCC 29521 by PCR. BSH gene was inserted into lactic acid bacteria expression vector pNZ8148 to construct the recombinant pNZ8148-BSH. The recombinant pNZ8148-BSH was transferred into lactic acid bacteria NZ9000 with electrotransformation method. And the recombinant which could express BSH protein was obtained. It was identified by SDS-PAGE electrophoresis and activity verification. The result could provide a rationale reference for expressing BSH in lactic acid bacteria.

Cloning and Expression of Bile Salt Hydrolase Gene from Lactobacillus plantarum M1-UVS29

We cloned and expressed bile salt hydrolase gene ofLactobacillus plantarum M1-UVS29 in Lactococcus lactis NZ9000 successfully. Gene-specific primers for amplification of L. plantarum bsh were designed by using sequence which availabled from GenBank. The production of PCR amplicon was confirmed by sequencing and cloned into pMD18-T vector, and then recombined into expression vector pNZ8148 and yielding vector pNZ8148-BSH, pNZ8148-BSH was transferred into Lactococcus lactis NZ9000. Sequencing indicated that the cloned bsh fragment contained 995 nucleotides, and shared 99.3% sequence homology with bsh gene from L. plantarum MBUL10. Cloned bsh fragment was successfully transduced into NICE expression system and confirmed by PCR and restriction digest. Recombinant BSH protein was analyzed by SDS-PAGE. The molecular weight of BSH protein was approximately 37 ku. Activity of the expressed protein was 0.77 μmol· min^-1. The successfully expressed proteins by genetic engineering technology made the function of lactic acid bacteria be abundant and laid the foundation for further researches into cholesterol-lowering lactic acid bacterium food and probiotics.

Effect of bile salts and bile acids on human gastric mucosal epithelial cells

Objective：To explore the effect of bile salt and bile acid on cultured eternalized human gastric mucosa epithelium GES-1 cells. Methods：Cultured eternalized human gastric mucosa epithelium GES-1 cells were treated with media containing 6 different kinds of bile salts and 3 different kinds of bile acids and their mixture with different concentrations： GCDC（glycochenodeoxychoμte）, GDC （glycodeoxychoμte）, GC（glycochoμte）, TCDC（taurochenodeoxychoμte）, TDC（taurodeoxychoμte）, TC （taurochoμte）, LCA （lithocholicacid）, CA（cholic acid）, DCA（deoxycholic acid）（50 μ mol/L,250 μ mol/L,500 μ mol/L,1000 μ mol/L）, DY（mixture of bile salts） and DS（mixture of bile acids）（250 μ mol/L,500 μ mol/L,1000 μ mol/L,1500 μ mol/L, 2000 μ mol/L）, in comparison with the control group（in normal media without bile salts and bile acids）. Cell proliferation was assessed by MTT（3-[4,5-Dimethylthiaolyl]-2,5- diphenyl-tetrazolium bromide） assay for 72 hours with different concentrations and the apoptotic cells were assayed by flow cytometry （FCM） with Annex V-FITC conjugated with propidium iodide（PI） staining for 24 hours with different concentrations（1500,2000 μt mol/L）. Results：There was no significant difference in morphology and cell proliferation in GC group after 24-72 h. Low concentration（50 μ mol/L） of GCDC, GDC, TCDC, TDC and TC accelerated gastric epithelial cell growth in a dosage-time dependent manner. At middle concentration （250-500 μ mol/L）, it showed positive effect after 24-48 h, while negative effect after 72 h. At high concentration（1000 μ mol/L）, it accelerated gastric epithelial cell growth after 24h and show consistent inhibition even leading to necrosis after 48-72 h. LCA and CA showed a positive effect on the concentration of 50 μ mol/L after 24-72 h, while 250-1000 μ mol/L showed a trend towards apoptosis after 24-72 h. At 50-500 μmol/L, DCA showed proliferation after 24 h and apoptosis after 48-72 h, but showed necrosis after 24-72 h at 1000 μmol/L. DY and DS could facilitate normal gastric mucosa epithelial cell growth at low concentration （250-500 μ mol/L）, however at 1000-2000 μ mol/L the trend shifted from apoptosis to necrosis. FCM with Annexin-V conjugated with PI staining revealed that GCDC, GDC, GC, TCDC, TDC, TC, LCA, CA, DCA, DY and DS induced apoptosis of human gastric mucosal epithelial cells. They were all significantly higher than that of the control（P 〈 0.05）, but there was no significant difference in GC group （P 〉 0.05）. The bile salts induced apoptosis in a time-dose-dependent manner. Conclusion：Our results suggested that bile acid and bile salt is the trigger of injury in human gastric mucosal epithelial cells.

Bile salt derivatives with novel skeleton from sea lamprey function as putative pheromone

OBJECTIVE The invasive sea lamprey(Petromyzon marinus)has devastated the ecosystem of the Laurentian Great Lakes.Application of pheromones to manipulate adult sea lamprey behavior is among the options considered for alternative sea lamprey control techniques.The male sea lamprey sex pheromone is hypothesized to be possess multiple functions through actions of multiple components,some of which have yet to be characterized.Our objective is to isolate and characterize the bioactive components from water conditioned with sexually mature male sea lamprey.METHODS The water conditioned with sexually mature male sea lamprey was extracted by solid phase extraction and concentrated in vacuo.The compounds were isolated by liquid chromatography and elucidated by spectrometry and spectroscopy.Their biological activities were evaluated by electro-olfactogram recordings and two-choice maze behavioral assays.RESULTS Five novel bile salts,petromyzene A and B and petromyzone A-C,have been characterized.Petromyzene A and B featured either a unique,rearranged side chain or a rare cis-11,12-diol on the steroidal B-ring.Petromyzone A-C represented three novel highly oxidized sulfated bile alcohols possessing different hydroxylation,oxidation,and double bond patterns,which exemplify the chemical diversity of bile salts.These five bile salts were potent odorants that stimulated the adult sea lamprey olfactory epithelium in a concentration dependent manner and showed detection thresholds between 10–13mol·L^(-1) and 10^(–11)mol·L^(-1)(paired t-test,P<0.05).Experiments in the two-choice maze showed that all isolated compounds induced behavioral responses in ovulated females.CONCLUSION The five novel compounds are likely additional components of pheromones released by sexually mature male sea lamprey,and may provide useful behavioral manipulation tools to be implemented with the integrated management of the destructive and invasive sea lamprey in the Laurentian Great Lakes.

<i>In-Vitro</i>Fermentation by Human Fecal Bacteria and Bile Salts Binding Capacity of Physical Modified Defatted Rice Bran Dietary Fiber

Defatted rice bran dietary fiber (DRBDF) was modified by micronization, ultrasound, microwave and extrusion cooking. We investigated the impacts of these physical treatments on the fermentation ability and bile salts binding capacity of DRBDF. In-vitro fermentation by human fecal bacteria of modified fibers showed that the major fermentation products were propionic, acetate and butyrate acid. Fermentation of extruded fiber gave the highest amounts of propionic and acetic acid 135.76 and 25.45 mmol/L respectively, while, the fermented product with microwaved fiber had the highest butyric acid content (10.75 mmol/L). The amount of short-chain fatty acid increased from 12 h to 24 h and propionic acid was the predominant. On the other hand,in-vitrobile salts binding showed that extruded fiber had higher affinity with sodium deoxycholate and sodium chenodeoxycholate (66.14% and 30.25% respectively) while microwaved fiber exhibited the highest affinity with sodium taurocholate (14.38%). In the light of obtained results we can affirmed that these physical treatments significantly improved the fermentation products and bile salts binding capacity of DRBDF. Extrusion compared to the other physical treatment methods used in this study has greatly and positively influenced the fermentation and bile binding capacity of DRBDF.

Denaturation studies on bovine serum albumin–bile salt system:Bile salt stabilizes bovine serum albumin through hydrophobicity

Protein denaturation is under intensive research, since it leads to neurological disorders of severe consequences. Avoiding denaturation and stabilizing the proteins in their native state is of great importance,especially when proteins are used as drug molecules or vaccines. It is preferred to add pharmaceutical excipients in protein formulations to avoid denaturation and thereby stabilize them. The present study aimed at using bile salts(BSs), a group of well-known drug delivery systems, for stabilization of proteins.Bovine serum albumin(BSA) was taken as the model protein, whose association with two BSs, namely sodium cholate(Na C) and sodium deoxycholate(Na DC), was studied. Denaturation studies on the preformed BSA-BS systems were carried out under chemical and physical denaturation conditions. Urea was used as the chemical denaturant and BSA-BS systems were subjected to various temperature conditions to understand the thermal(physical) denaturation. With the denaturation conditions prescribed here,the data obtained is informative on the association of BSA-BS systems to be hydrophobic and this effect of hydrophobicity plays an important role in stabilizing the serum albumin in its native state under both chemical and thermal denaturation.

Selective binding of bile salts by β-cyclodextrin derivatives with appended quinolyl arms

Two β-cyclodextrin derivatives bearing appended quinolyl and isoquinolyl arms,i.e.mono-（6-quinolyl- 6-deoxy）-β-cyclodextrin（1） and mono-（6-isoquinolyl-6-deoxy）-β-cyclodextrin（2） were synthesized in satisfactory yields and fully characterized.Their original conformations and binding behaviors toward four bile salt guests,that is,sodium cholate（CA）,sodium deoxycholate（DCA）,sodium glycocholate （GCA）,and sodium taurocholate（TCA）,were investigated by means of fluorescence,circular dichroism and 2D NMR spectroscopy.The study of solution structures revealed that both quinolyl and isoquinolyl arms were located outside the cyclodextrin cavity.The results obtained from the fluorescence titrations showed that the binding abilities of hosts 1 and 2 with selected bile salts varied in an order of DCA 〉 CA 〉 GCA.The selective binding of hosts toward bile salt guests was discussed from the viewpoints of induced-fit and multiple binding.

Gastric and small intestinal lipid digestion kinetics as affected by the gradual addition of lipases and bile salts

We studied the effect of evolving from static in vitro digestion conditions towards the gradual addition of essential digestive compounds for lipid digestion.Two oil-in-water emulsions were considered:a low(5%w/w)and high(20%w/w)triolein-based emulsion with identical surfactant-to-oil ratio(0.2).Emulsions were subjected to in vitro static digestion conditions or gradual gastric lipase addition,gradual pancreatic lipase addition,and/or gradual bile salt addition.For these three latter cases,similar amounts of gastric/pancreatic lipase and/or bile salts were provided as in the static case,however divided over 4 doses added during the first 30 min of each digestive phase.For the low-lipid emulsion,gradually adding lipases and bile salts did not significantly affect lipolysis kinetics.This can be related to the sufficient amounts of digestive compounds present even in the smallest initial dose.For the high-lipid emulsion,the gradual addition of bile salts significantly reduced the lipolysis rate.Bile salts are essential to remove lipid digestion products from the interface and thus allow the continuation of the lipid digestion process at the interface.Oppositely,the lipolysis extent after 2 h of small intestinal phase was not significantly influenced by the digestion approach.This is again explained by the simple nature of the emulsions studied and the excess of lipase even in the smallest initial dose.Overall,this work showed that evolving towards more(semi-)dynamic digestion conditions can impact(lipid)digestion kinetics,even for relatively simple food compositions,and is of interest to obtain more physiological relevant digestion kinetics.

Effects of bile salts and divalent cations on the adsorption of norfloxacin by agricultural soils

The effects of bile salts （sodium cholate and sodium deoxycholate, 0-20 mmol/L）, divalent cations （Ca^2＋, Mg^2＋, Cu^2＋ and Zn^2＋, 0-20 mmol/L） or pH （3.0-10.0） on the adsorption of norfloxacin by three selected soils （Paddy_H, Paddy_G and Red_J） were systematically studied. Soil adsorption of norfloxacin follows a pseudo second-order kinetics model, and the maximum adsorption capacity has been determined from the nonlinear fit of the Langmuir isotherm model to be 88.8, 88.1 and 63.0 μmol/g for the adsorption onto Paddy_H, Paddy_G and Red_J, respectively. The results indicate that norfloxacin has a high adsorption affinity for the agricultural soils tested and that the organic content of these soils have at least a slight influence on this adsorption. The adsorption of norfloxacin to soils was strongly dependent on pH and exhibited a maximum at approximately pH 6. The presence of divalent cations prominently suppressed the adsorption of norfloxacin by paddy soils, which followed an order of Cu^2＋ 〉 Mg^2＋ 〉 Ca^2＋ 〉 Zn^2＋, and by red soil, which followed an order of Cu^2＋ 〉 Zn^2＋ 〉 Ca^2＋ 〉 Mg^2＋. The adsorption of norfloxacin （by the soils studied） sharply decreased as the amount of bile salts was increased. For uncharged norfloxacin at environmentally relevant pH values, such factors as soil type, exogenous divalent cations and macromolecules significantly altered the environmental fate and transport of norfloxacin between aquatic and soil interfaces.

深共熔溶剂提取桑黄黄酮及体外降血糖、结合胆酸盐能力分析

从7种深共熔溶剂中筛选最适合用于桑黄黄酮化合物提取的溶剂体系,采用响应面优化深共熔溶剂提取桑黄黄酮的工艺,筛选最佳黄酮回收树脂,并分析其体外降血糖和结合胆酸盐能力。试验结果表明,乙二醇与氯化胆碱组成的深共熔溶剂体系最适合提取桑黄黄酮,最佳提取工艺为深共熔溶剂含水率30%,乙二醇与氯化胆碱的物质的量比为8.21∶1,料液比=50∶1(mg∶m L),提取温度64℃,提取时间43 min,在此条件下,桑黄黄酮得率为(8.85±0.34)%,较体积分数为60%乙醇提取的得率提高了2.02倍。9种大孔树脂中,AB-8型大孔树脂对桑黄黄酮的回收率最高,为85.25%;纯化后桑黄黄酮的纯度提高了6.12倍。桑黄黄酮抑制α-淀粉酶和α-葡萄糖苷酶活性的IC_(50)值分别为1.19 mg/m L和1.28 mg/m L,结合牛磺胆酸钠和甘氨胆酸钠的IC_(50)值分别为0.62 mg/m L和1.19 mg/m L,表明桑黄黄酮具有较强的调节血糖和血脂能力。实验表明,深共熔溶剂结合大孔吸附树脂富集回收可以绿色高效地从桑黄中提取黄酮类化合物。

Alleviative effects of Bacillus coagulans strains on irritable bowel syndrome-unraveling strain specificity through physiological and genomic analysis

The high intraspecies heterogeneity of Baciillus coagulans leads to significant phenotypic differences among different strains.Thus,6 B.coagulans strains were tested in the present study using an irritable bowel syndrome(IBS)animal model to determine whether the IBS-alleviating effects of B.coagulans strains are strain-specific.The results of this study showed that the ingestion of B.coagulans GBI-30,6086,and B.coagulans CCFM1041 significantly alleviated IBS symptoms in mice.In contrast,other B.coagulans strains showed no or limited alleviating effects on IBS symptoms.According to our experimental results,the two main common features of these strains were as follows:1)The resistance of vegetative cells to bile salts,and 2)ability to synthesize specific lipids and secondary metabolites.Screening strains based on these two indicators may greatly reduce costs and provide a basis for mining new functional B.coagulans strains.Our results also suggest that administration of B.coagulans could significantly regulate microbiota dysbiosis in animal models.Moreover,the close relationships between the gut microbiota,gut microbiota metabolites,and IBS were further confirmed in this study.

Influence of gut microbiome composition on effect variations of anti-cholesterol treatment among individual mice

This study investigated if the variation in the effect of anti-cholesterol(AC)treatment on individual mice are related to gut microbiome composition.The bile salt hydrolase(BSH)activity of 23 commercial fermented milk products was examined to select a fermented milk product for AC treatment.Mice were fed to different diets for 6 weeks:high-fat(60%of total calories from fat;D1),high-dietary fibre(20%cellulose;D2),and low-fat(17.2%of total calories from fat;D3)diets to change their gut microbiomes.Subsequently,faecal microbiome was transplanted(FMT)into mice treated with high cholesterol diet contained 2%cholesterol,followed by AC or non-AC(sterile tap water,STW)treatments.Control groups with normal(NC)and highcholesterol diets(PC)were prepared for both AC and STW treatment.All experimental groups were subjected to serum and liver cholesterol,cholesterol metabolism-related(CMR)gene expression,and intestinal microbiome analyses.D3-FMT mice showed the most significant enhancements in cholesterol ratio and decreased hepatic cholesterol levels with AC treatment.Moreover,upregulation of the Cyp7a1 gene expression was observed in this group.Furthermore,the intestinal microbiome analysis indicated higher abundances of BSH-producing Eubacterium,Bifidobacterium,and Parabacteroides in the D3-FMT+AC group compare to others,potentially contributing to increased bile acid synthesis.

高产胆盐水解酶乳杆菌的筛选及对新生大鼠黄疸的防治作用

为筛选高产胆盐水解酶的乳杆菌,探究其对新生儿黄疸的防治作用。采用添加了25 U/mL制霉菌素的LBS选择性培养基,从健康新生儿粪便和母乳中筛选乳杆菌并鉴定种类;以鼠李糖乳杆菌LGG为阳性对照,体外评估菌株的益生菌特性;利用盐酸苯肼诱导新生SD大鼠黄疸模型,通过分析血清胆红素水平和肝脏组织的损伤情况,以及肝脏炎症因子、核转录因子的相对表达水平,探究高产胆盐水解酶乳杆菌对新生大鼠黄疸的防治作用及机制。结果表明,来自婴儿粪便的格氏乳杆菌FWJL-5在体外具良好的益生特性,并且产胆盐水解酶能力优于LGG,能够显著缓解新生大鼠胆红素水平升高、肝脏组织肿胀和溶血症状,减少肝脏损伤中肝酶的释放,抑制促炎因子的分泌,促进UGt1A1和上游核转录因子孕烷X受体(pregnane X receptor,pXR)、法尼醇X受体(farnesol X receptor,FXR)的表达。综上所述,婴儿粪便来源的格氏乳杆菌FWJL-5可通过上调核受体FXR/pXR促进UGt1A1表达以调节肝脏胆红素代谢,从而减轻新生大鼠黄疸症状,本研究可为格氏乳杆菌防治新生儿黄疸提供新思路。

High-fat-induced intestinal permeability dysfunction associated with altered fecal bile acids

AIM： To investigate whether high-fat-feeding is associ- ated with increased intestinal permeability via altera- tions in bile acid metabolism.METHODS： Male C57BI/6J mice were fed on a high-fat （n = 26） or low-fat diet （n = 24） for 15 wk. Intestinal permeability was measured from duodenum, jejunum, ileum and colon in an Ussing chamber system using 4 kDa FITC-labeled dextran as an indicator. Fecal bile ac- ids were analyzed with gas chromatography. Segments of jejunum and colon were analyzed for the expression of farnesoid X receptor （FXR） and tumor necrosis factor

血浆胆汁酸在肝肿瘤中的差异分析及应用价值

目的分析原发性肝癌与转移性肝癌患者血浆胆汁酸水平及其与临床指标的相关性,并评估其联合AFP对于原发性肝癌的诊断价值。方法纳入2020年8月—2021年9月于上海中医药大学附属曙光医院就诊且具有组织病理学及影像学明确的75例原发性肝癌患者及79例转移性肝癌患者,采集外周血并分别分离血清及血浆,比色法及色谱法检测生化指标,电化学发光免疫分析法检测肿瘤标志物水平,高效液相色谱-串联质谱法检测胆汁酸含量。符合正态分布的计量资料两组间比较采用成组t检验,不符合正态分布的计量资料两组间比较采用Mann-Whitney U检验。相关性检验采用Spearman相关分析。临床诊断效能采用受试者工作特征曲线(ROC曲线)评估。结果原发性肝癌组患者TC、TG、LDL-C及载脂蛋白(Apo)B显著低于转移性肝癌组患者,差异均具有统计学意义(U值分别为1598、1255、909、889,P值均<0.05)。原发性肝癌组患者AFP显著高于转移性肝癌组患者,癌胚抗原(CEA)显著低于转移性肝癌组患者,差异均具有统计学意义(U值分别为1873、926,P值均<0.05)。原发性肝癌组患者总胆汁酸(TBA)、胆酸(CA)、鹅脱氧胆酸(CDCA)、熊脱氧胆酸(UDCA)、牛磺胆酸(TCA)、牛磺鹅脱氧胆酸(TCDCA)、甘氨胆酸(GCA)、甘氨鹅脱氧胆酸(GCDCA)、牛磺熊脱氧胆酸(TUDCA)、甘氨熊脱氧胆酸(GUDCA)均显著高于转移性肝癌患者,脱氧胆酸(DCA)显著低于转移性肝癌患者,差异均具有统计学意义(P值均<0.05)。总人群中TBA、CDCA、GCA、GCDCA、GUDCA、TCA、TCDCA及TUDCA含量与AFP水平呈显著正相关(P值均<0.05)。原发性肝癌患者中GCA、TCA、TCDCA及TUDCA含量与AFP水平呈显著正相关(P值均<0.05)。AFP+TCA+GCA+TCDCA联合检测的AUC为0.822(95%CI:0.746~0.898,P<0.0001),效能最高。结论原发性肝癌与转移性肝癌患者血浆胆汁酸含量具有显著差异,差异性胆汁酸与肝损伤及AFP升高密切相关,联合AFP对原发性肝癌具有更优的临床诊断价值。