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 w...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.展开更多
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 syndr...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.展开更多
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 ...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展开更多
AIM To investigate the morphologic changes of the myocardium and its relationship to serum bile acids in obstructive jaundice. METHODS Part Ⅰ: 35 rats were randomly assigned to three groups: Group Ⅰ (BDL1, n =...AIM To investigate the morphologic changes of the myocardium and its relationship to serum bile acids in obstructive jaundice. METHODS Part Ⅰ: 35 rats were randomly assigned to three groups: Group Ⅰ (BDL1, n =11), the common bile duct (CBD) was ligated and severed and killed after one week. Group Ⅱ (BDL2, n =11), the CBD was ligated and severed and killed after two weeks. Group Ⅲ (SO, n =13), the CBD was simply isolated. The hearts were taken for morphologic studies and blood was taken to determine the total serum bile acids (TAB). Part Ⅱ: 13 rats received gastric intubation of 10% 4ml/kg of sodium cholate, and their serum TBA and the morphologic changes of the heart were examined. RESULTS One to two weeks after the CBD was ligated and severed, the mitochondrium of the myocardium was damaged and the serum TBA obviously increased. When the rats were administered sodium cholate to make their peak blood concentration close to the average blood concentration in BDL2, their myocardium was damaged in a similar degree. CONCLUSION The myocardium was damaged in obstructive jaundice and the endogenous bile acids was one of the factors.展开更多
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) gen...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.展开更多
IM To analyze serum bile acids and biliary lipids of patients with cholesterol gallstone(CS) and explore the relationship between deoxycholic acid (DCA) and CS disease.METHODS Analysis of bile acids in serum was do...IM To analyze serum bile acids and biliary lipids of patients with cholesterol gallstone(CS) and explore the relationship between deoxycholic acid (DCA) and CS disease.METHODS Analysis of bile acids in serum was done with gaschromatography in two groups: CS group (n=151) and control group (n=256). Serum bile acids and biliary lipids were also studied in 90 matched samples..RESULTS The serum DCA was 0955μmol/L±0078μmol/L in CS group, which was more than that of control group (0696μmol/L±0047μmol/L), P<001. The ratio of DCA/chenodeoxycholic acids (CDCA) was 176±030 in CS group, about two times that in control group (092±014). The mole percent of DCA in bile was positively related to cholesterol saturation index (CSI) (P<001) and the mole percent of CDCA in bile negatively to CSI (P=001). There was correlation between the mole percent of DCA, CDCA and cholic acid in bile and in serum.CONCLUSION It is suggested that DCA is lithogenic and the increased amount of DCA or the ratio of DCA/CDCA in serum may be one of the features of cholesterol gallstone patients.展开更多
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...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.展开更多
This review considers the physiological and molecular biochemical mechanisms of bile formation.The composition of bile and structure of a bile canaliculus,biosynthesis and conjugation of bile acids,bile phospholipids,...This review considers the physiological and molecular biochemical mechanisms of bile formation.The composition of bile and structure of a bile canaliculus,biosynthesis and conjugation of bile acids,bile phospholipids,formation of bile micellar structures,and enterohepatic circulation of bile acids are described.In general,the review focuses on the molecular physiology of the transporting systems of the hepatocyte sinusoidal and apical membranes.Knowledge of physiological and biochemical basis of bile formation has implications for understanding the mechanisms of development of pathological processes,associated with diseases of the liver and biliary tract.展开更多
基金the Clinical Key Programs of Ministry of Public Health(No.20012130)
文摘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.
基金supported by the Natural Science Foundation of Jiangsu Province(BK20200084)the National Natural Science Foundation of China(31871773 and 31820103010)+1 种基金the Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province and Key Talents Project of“Strengthening Health through Science and Education”of Wuxi Health and Family Planning Commission(ZDRC039)Top Talents Project of“Six-one Project”for High-level Health Talents in Jiangsu Province(LGY2018016)。
文摘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.
基金Supported by The Foundation for Nutrition Research and the Finnish Funding Agency of Technology and Innovation
文摘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
文摘AIM To investigate the morphologic changes of the myocardium and its relationship to serum bile acids in obstructive jaundice. METHODS Part Ⅰ: 35 rats were randomly assigned to three groups: Group Ⅰ (BDL1, n =11), the common bile duct (CBD) was ligated and severed and killed after one week. Group Ⅱ (BDL2, n =11), the CBD was ligated and severed and killed after two weeks. Group Ⅲ (SO, n =13), the CBD was simply isolated. The hearts were taken for morphologic studies and blood was taken to determine the total serum bile acids (TAB). Part Ⅱ: 13 rats received gastric intubation of 10% 4ml/kg of sodium cholate, and their serum TBA and the morphologic changes of the heart were examined. RESULTS One to two weeks after the CBD was ligated and severed, the mitochondrium of the myocardium was damaged and the serum TBA obviously increased. When the rats were administered sodium cholate to make their peak blood concentration close to the average blood concentration in BDL2, their myocardium was damaged in a similar degree. CONCLUSION The myocardium was damaged in obstructive jaundice and the endogenous bile acids was one of the factors.
基金Supported by 863 Projects (2008AA10Z311)National Science and Technology Support Projects (2009BADB9B06)+1 种基金Started Post-doctoral Research Grant of Heilongjiang Province (LBH-Q07023)Harbin Technological Innovation of Special Funds (2007RFQXN020)
文摘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.
文摘IM To analyze serum bile acids and biliary lipids of patients with cholesterol gallstone(CS) and explore the relationship between deoxycholic acid (DCA) and CS disease.METHODS Analysis of bile acids in serum was done with gaschromatography in two groups: CS group (n=151) and control group (n=256). Serum bile acids and biliary lipids were also studied in 90 matched samples..RESULTS The serum DCA was 0955μmol/L±0078μmol/L in CS group, which was more than that of control group (0696μmol/L±0047μmol/L), P<001. The ratio of DCA/chenodeoxycholic acids (CDCA) was 176±030 in CS group, about two times that in control group (092±014). The mole percent of DCA in bile was positively related to cholesterol saturation index (CSI) (P<001) and the mole percent of CDCA in bile negatively to CSI (P=001). There was correlation between the mole percent of DCA, CDCA and cholic acid in bile and in serum.CONCLUSION It is suggested that DCA is lithogenic and the increased amount of DCA or the ratio of DCA/CDCA in serum may be one of the features of cholesterol gallstone patients.
基金supported by the National Research Foundation of Korea(NRF)grant funded by the Korea government(MSIT)(2019R1A2C2004356).
文摘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.
文摘This review considers the physiological and molecular biochemical mechanisms of bile formation.The composition of bile and structure of a bile canaliculus,biosynthesis and conjugation of bile acids,bile phospholipids,formation of bile micellar structures,and enterohepatic circulation of bile acids are described.In general,the review focuses on the molecular physiology of the transporting systems of the hepatocyte sinusoidal and apical membranes.Knowledge of physiological and biochemical basis of bile formation has implications for understanding the mechanisms of development of pathological processes,associated with diseases of the liver and biliary tract.