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Upregulation of 25-hydroxyvitamin D_3-1α-hydroxylase by butyrate in caco-2 cells

World Journal of Gastroenterology 2005年第45期11卷 7136-7141页

作者： Oliver Schr(o|¨)der Sinan Turak Carolin Daniel Tanja Gaschott Jürgen Stein Ⅰst Department of Internal Medicine

AIM： To investigate the possible involvement of 25-hydroxyvitamin D3-1cx-hydroxylase [1α-25（OH）2D3] in butyrate-induced differentiation in human intestinal cell line caco-2 cells. METHODS： caco-2 cells were incubated either with 3 mmol/L butyrate and 1 umol/L 25（OH）2D3 or with 1 umol/L 1α-25（OH）2D3 for various time intervals ranging from 0 to 72 h. Additionally, cells were co-incubated with butyrate and either 25（OH）2D3 or 1α-25（OH）2D3. 1α-25（OH）2D3 mRNA was determined semi-quantitatively using the fluorescent dye PicoGreen. Immunoblotting was used for the detection of 1α-25（OH）2D3 protein. Finally, enzymatic activity was measured by ELISA. RESULTS： Both butyrate and 1α-25（OH）2D3 stimulated differentiation of caco-2 cells after a 48 h incubation period, while 25（OH）2D3 had no impact on cell differentiation. Synergistic effects on differentiation were observed when cells were co-incubated with butyrate and vitamin D metabolite. Butyrate transiently upregulated 1α-25（OH）2D3 mRNA followed by a timely delayed protein upregulation. Coincidently, enzymatic activity was enhanced significantly. The induction of the enzyme allowed for comparable differentiating effects of both vitamin D metabolites. CONCLUSION： Our experimental data provide a further mechanism for the involvement of the vitamin D signaling pathway in colonic epithelial cell differentiation by butyrate. The enhancement of 1α-25（OH）2D3 followed by antiproliferative effects of the vitamin D prohormone in the caco-2 cell line suggest that 25（OH）2D3 in combination with butyrate may offer a new therapeutic approach forthe treatment of colon cancer.

关键词： 25-Hydroxyvitamin D3-1α-hydroxylase Butyrate caco-2 cells Colon cancer Differentiation Vitamin D

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Effects of extracellular iron concentration on calcium absorption and relationship between Ca^(2+) and cell apoptosis in caco-2 cells

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World Journal of Gastroenterology 2005年第19期11卷 2916-2921页

作者： LiWang QingLi Xiang-LinDuan Yan-ZhongChang TheKeyLaboratoryofAnimalPhysiology BiochemistryandMolecularBiologyofHebeiProvinceLifeScienceCollegeHebeiNormalUniversityShijiazhuang050016HebeiProvinceChina

AIM: To determine the method of growing small intestinal epithelial cells in short-term primary culture and to investigate the effect of extracellular iron concentration ([Fe3+]) on calcium absorption and the relationship between the rising intracellular calcium concentration ([Ca2+]i) and cell apoptosis in human intestinal epithelial caco-2 cells. METHODS: Primary culture was used for growing small intestinal epithelial cells. [Ca2+]i was detected by a confocal laser scanning microscope. The changes in [Ca2+]i were represented by fluorescence intensity (FI). The apoptosis was evaluated by flow cytometry. RESULTS: Isolation of epithelial cells and preservation of its three-dimensional integrity were achieved using the digestion technique of a mixture of collagenase Ⅺ and dispase Ⅰ. Purification of the epithelial cells was facilitated by using a simple differential sedimentation method. The results showed that proliferation of normal gut epithelium in vitro was initially dependent upon the maintenance of structural integrity of the tissue. If 0.25% trypsin was used for digestion, the cells were severely damaged and very difficult to stick to the Petri dish for growing. The Fe3+ chelating agent desferrioxamine (100, 200 and 300 μmol/L) increased the FI of caco-2 cells from 27.50±13.18 (control, n = 150) to 35.71±13.99 (n = 150, P2.19±35.40 (n = 150, P211.34±29.03 (n = 150,Pcaco-2 cells treated by ferric ammonium citrate (FAC, a Fe3+ donor; 10, 50 and 100 μmol/L). The FI value of caco-2 cells treated by FAC was 185.85±33.77 (n = 150, P22.73±58.47 (n = 150, P29±19.82 (n= 150,P23187(0.1,1.0 and 10 μmol/L) increased the FI of caco-2 cells from 40.45±13.95 (control, n = 150) to 45.19±21.95 (n = 150, P29 (n = 150, P<0.01) and

关键词： Iron calcium absorption Cell apoptosis caco-2 cells

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Gastric digestion of pea ferritin and modulation of its iron bioavailability by ascorbic and phytic acids in caco-2 cells

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World Journal of Gastroenterology 2007年第14期13卷 2083-2088页

作者： Satyanarayana Bejjani Raghu Pullakhandam Ravinder Punjal K Madhavan Nair Micronutrient Research Group National Institute of Nutrition Indian Council of Medical ResearchJamai OsmaniaHyderabad 500007India

AIM： To understand the digestive stability and mechanism of release and intestinal uptake of pea ferritin iron in caco-2 cell line model.METHODS： Pea seed ferritin was purified using salt fractionation followed by gel filtration *** bioavailability of ferritin iron was assessed using coupled in vitro digestion/caco-2 cell model in the presence or absence of ascorbic acid and phytic ***-2 cell ferritin formation was used as a surrogate marker of iron uptake. Structural changes of pea ferritin under simulated gastric pH were characterized using electrophoresis, gel filtration and circular dichroism spectroscopy.RESULTS： The caco-2 cell ferritin formation was significantly increased （P 〈 0.001） with FeSO4 （19.3±9.8 ng/mg protein） and pea ferritin （13.9 ± 6.19 ng/mg protein） compared to the blank digest （3.7 ± 1.8 ng/mg protein）. Ascorbic acid enhanced while phytic acid decreased the pea ferritin iron bioavailability. However,either in the presence or absence of ascorbic acid, the ferritin content of caco-2 cells was significantly less with pea ferritin than with FeSO4. At gastric pH, no band corresponding to ferritin was observed in the presence of pepsin either on native PAGE or SDS-PAGE. Gel filtration chromatography and circular dichroism spectroscopy revealed a pH dependent loss of quaternary and secondary structure.CONCLUSION： Under gastric conditions, the iron core of pea ferritin is released into the digestive medium due to acid induced structural alterations and dissociation of protein. The released iron interacts with dietary factors leading to modulation of pea ferritin iron bioavailability,resembling the typical characteristics of non-heme iron.

关键词： Pea ferritin Bioavailability caco-2 cells Phytic acid Ascorbic acid

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Impact of in vitro gastrointestinal digestion on rabbiteye blueberry anthocyanins and their absorption efficiency in caco-2 cells

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Food Bioscience 2023年第2期52卷 622-629页

作者： Daniela D.Herrera-Balandrano Jing Wang Zhi Chai Xiaoxiao Zhang Jialei Wang Ning Wang Wuyang Huang School of Life Sciences Nantong UniversityNantong226019PR China Institute of Agro-Product Processing Jiangsu Academy of Agricultural SciencesNanjing210014PR China College of Chemical Engineering Nanjing Forestry UniversityNanjing210037PR China School of Food and Biological Engineering Jiangsu University212013ZhenjiangPR China Central Laboratory Jiangsu Academy of Agricultural SciencesNanjing210014PR China

Blueberries are rich in anthocyanins,which have numerous nutritional and pharmacological *** this study,the absorption and stability of rabbiteye blueberry anthocyanins extract(BAE)were assessed using in vitro digestion and caco-2 cell ***,similar patterns were found through different phases of simulated in vitro digestion,however,the total anthocyanin content(TAC)of BAE significantly *** all thirteen anthocyanins from BAE,the highest bioaccessibility of>50%was found in Cyanidin-3-glucoside(Cy-3-glc),Petunidin-3-galactoside,(Pt-3-gal),and Cyanidin-3-arabinoside(Cy-3-ara).However,malvidin glycosides represented around 55%of TAC with 27-35%*** absorption caco-2 cell monolayers assay indicated that blueberry anthocyanins with higher hydrophobicity are more efficiently *** addition,the structure of glycosides impacts the absorption and transport efficiencies of *** study provides data on the stability and absorption of blueberry anthocyanins,that can be used to explore the underlying mechanism of BAE absorption.

关键词： Blueberry Anthocyanins In vitro digestion caco-2 cells Transport Absorption

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Overloading of differentiated caco-2 cells during lipid transcytosis induces glycosylation mistakes in the Golgi complex

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BIOCELL 2021年第3期45卷 773-783页

作者： GALINA N.DENISOVA IVAN D.DIMOV ANNA V.ZAITSEVA LINARD J.ARTIUX ALEXANDER A.MIRONOV NATALIA R.KARELINA Department of Anatomy Saint Petersburg State Paediatric Medical UniversityPetersburg194100Russia Department of Biochemistry Saint Petersburg State Paediatric Medical UniversityPetersburg194100Russia Department of Heart and Vessel Surgery Saint Petersburg State Paediatric Medical UniversityPetersburg194100Russia The FIRC Institute of Molecular Oncology Milan20139Italy

Overloading the intestine enterocytes with lipids induced alteration of the Golgi complex(GC;Sesorova et al.,2020)and could cause glycosylation ***,using differentiated caco-2 cells with the established 0[I]blood group phenotype(no expression of the blood antigens A and B[AgA,AgB]under normal conditions)as a model of human enterocytes we examined whether the overloading of these cells with lipids could cause errors in the Golgi-dependent *** demonstrated that under these conditions,there were alterations of the GC and the appearance of lipid droplets in the *** cells produced AgA and *** suggested that after overloading of enterocytes with lipids,AgA were mistakenly synthesized in individual enterocytes by the Golgi *** mistakes could explain why in the absence of AgA and AgB antibodies against them exist in the blood.

关键词： Golgi Blood group antigens Glycosylation errors Enterocyte Lipid transcytosis caco-2 cells

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Inhibitory effects of YCW and MOS from Saccharomyces cerevisiae on Escherichia coli and Salmonella pullorum adhesion to caco-2 cells

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Frontiers in Biology 2017年第5期12卷 370-375页

作者： Xiaoqing Xu Yu Qiao Qing Peng Long Gao Bo Shi Key Laboratory for Feed Biotechnology of the Ministry of Agriculture Feed Research Institute Chinese Academy of Agricultural SciencesBeijing 100081 China

BACKGROUND： For many years, yeast cell walls （YCW） and mannan oligosaccharides （MOS） have been used as alternatives to antibiotics and health feed additives to enhance the growth performance and health of food animals. In the present study, the inhibitory effects of YCW and MOS on the adhesion of enteropathogenic bacteria to intestinal epithelial cells were tested. METHODS： YCW and MOS were extracted from Saccharomyces cerevisiae （XM 0315）, and the morphology of YCW and MOS bound to pathogenic bacteria was observed by scanning electron microscopy （SEM）. Real-time fluorescent quantitative PCR was used to quantitatively analyze the effects of YCW and MOS on the adhesion of Escherichia coli （CVCC3367） and Salmonella pullorum （CVCC520） to caco-2 cells. RESULTS： The results showed that YCW inhibited E. coli and S. pullorum binding to caco-2 cells by 95% and 74%, respectively, whereas MOS prevented E. coli and S. pullorum binding by 67% and 50%, respectively. CONCLUSIONS： These data suggest that YCW has a stronger ability than MOS to inhibit pathogenic bacteria from adhering to caco-2 cells in vitro.

关键词： YCW MOS Escherichia coli Salmonella pullorum caco-2 cells

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Antioxidant Activity and Protective Effects of Alcalase-Hydrolyzed Soybean Hydrolysate in Human Intestinal Epithelial caco-2 cells

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大豆科技 2019年第S1期 54-65,75页

作者： Zhang Qiaozhi Tong Xiaohong Sui Xiaonan Wang Zhongjiang Qi Baokun Li Yang Jiang Lianzhou 不详

Soybeans are known as a promising source of bioactive ***,knowledge on the antioxidant behaviors of soybean protein hydrolysate(SPH)in the human intestinal epithelium is *** this study,SPH was prepared with Alcalase and subsequently ultrafiltered into four peptide fractions as SPH-I(10 kDa).The antioxidant properties of SPH and membrane fractions were investigated using different chemical assays and their protective effects against oxidative stress were evaluated using H2 O2-stressed human intestinal caco-2 *** showed that SPH-I exhibited the strongest 2,2-diphenyl-1-picrylhydrazyl(DPPH)radical scavenging activity(IC50=2.56 mg/m L)and reducing capacity while SPH-III had the best metal ion-chelating activity(IC50=0.29 mg/m L).Both SPH and the peptide fractions dose-dependently suppressed intracellular reactive oxygen species(ROS)accumulation induced by H2O2 in caco-2 cells,but the strongest inhibitory effect was observed for *** acid(AA)results revealed that SPH-I was rich in hydrophobic and antioxidant AAs,which could contribute to its stronger antioxidant ***,SPH-I protected caco-2 cells from H2O2-induced oxidative stress via inhibiting lipid peroxidation and stimulating antioxidant enzyme *** results suggest that SPH-I and constitutive peptides can be beneficial ingredients with antioxidant properties and protective effects against ROS-mediated intestinal injury.

关键词： Soybean protein hydrolysate Antioxidant activity Alcalase Oxidative stress Cytoprotective effect caco-2 cells

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Simple prediction of caco-2 cells permeability for flavone aglycones

Simple prediction of Caco-2 cells permeability for flavone a...

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第九届全国药物和化学异物代谢学术会议

作者： XU Shi,CHEN Diao,WANG Zhan-guo,JIANG Xue-hua,LAN Ke~* Key Laboratory of Drug Targeting and Novel Drug Delivery Systems of the Education Ministry,West China School of Pharmacy,Sichuan University,Chengdu 610041,Sichuan,China

Flavonoids were well-known for their considerable in vitro bioactivities and poor oral *** better understand the biological fate of flavonoids,virtual combinatorial chemistry,3D molecular fields and Partial Least Square-Data Discrimination(PLSDA) analysis were utilized to virtually predict the caco-2 cells permeability of flavone *** on our preliminary results,two basic rules were concluded.(1) Hydroxylations commonly hinder the caco-2 cells permeation of flavone aglycones,while hydroxyl-methylations help recover caco-2 cells penetration;(2) Either hydroxylations or methoxylations on the B-ring of flavone aglycones possibly the most hinders the caco-2 cells *** though the reported data of a specific flavone aglycone was highly variable in different publications, rule 1 had been greatly approved by reports with purposeful study ***,to date,there was not enough data to support rule 2,whose rationality has to be discussed by further experiments with purposeful study design.

关键词： permeability caco-2 cells structure-activity relationships computational models antioxidants

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Exogenous sphingomyelinase causes impaired intestinal epithelial barrier function

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World Journal of Gastroenterology 2007年第39期13卷 5217-5225页

作者： Jürgen Bock Gerhard Liebisch Joachim Schweimer Gerd Schmitz Gerhard Rogler Department of Internal Medicine I University of RegensburgGermany Institute for Clinical Chemistry University of Regensburg Germany

AIM: To test the hypothesis that hydrolysis of sphingomyelin to ceramide changes the composition of tight junctions (TJs) with increasing permeability of the intestinal epithelium. METHODS: Monolayers of caco-2 cells were used as an in vitro model for the intestinal barrier. Permeability was determined by quantifi cation of transepithelialflux and transepithelial resistance. Sphingolipid-rich membrane microdomains were isolated by a discontinuous sucrose gradient and characterized by Western-blot. Lipid content of microdomains was analysed by tandem mass spectrometry. Ceramide was subcellularly localized by immunofluorescent ***: Exogenous sphingomyelinase increased transepithelial permeability and decreased transepithelial resistance at concentrations as low as 0.01 U/mL. Lipid analysis showed rapid accumulation of ceramide in the membrane fractions containing occludin and claudin-4, representing TJs. In these fractions we observed a concomitant decrease of sphingomyelin and cholesterol with increasing concentrations of ceramide. Immunofluorescent staining confirmed clustering of ceramide at the sites of cell-cell contacts. Neutralization of surface ceramide prevented the permeability-increase induced by platelet activating factor. CONCLUSION: Our findings indicate that changes in lipid composition of TJs impair epithelial barrier functions. Generation of ceramide by sphingomyelinases might contribute to disturbed barrier function seen in diseases such as inflammatory, infectious, toxic or radiogenic bowel disease.

关键词： Ceramide Cholesterol Tight-junction caco-2 cells Permeability Inflammatory bowel disease

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Comparative permeability of three saikosaponins and corresponding saikogenins in caco-2 model by a validated UHPLC-MS/MS method

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Journal of Pharmaceutical Analysis 2021年第4期11卷 435-443页

作者： Siqi Ren Jingjing Liu Yunwen Xue Mei Zhang Qiwei Liu Jie Xu Zunjian Zhang Rui Song Key Laboratory of Drug Quality Control and Pharmacovigilance China Pharmaceutical UniversityNanjing210009China State Key Laboratory of Natural Medicine China Pharmaceutical UniversityNanjing210009China

Saikosaponins(SSs)are the main active components extracted from Bupleuri Radix(BR)which has been used as an important herbal drug in Asian countries for thousands of *** has been reported that the intestinal bacteria plays an important role in the in vivo disposal of oral *** the deglycosylated derivatives(saikogenins,SGs)of SSs metabolized by the intestinal bacteria are speculated to be the main components absorbed into the blood after oral administration of SSs,no studies have been reported on the characteristics of SGs for their intestinal absorption,and those for SSs are also ***,a rapid UHPLC-MS/MS method was developed to investigate and compare the apparent permeability of three common SSs(SSa,SSd,SSb2)and their corresponding SGs(SGF,SGG,SGD)through a bidirectional transport experiment on caco-2 cell monolayer *** method was validated according to the latest FDA guidelines and applied to quantify the six analytes in transport medium samples extracted via liquid-liquid extraction(LLE).The apparent permeability coefficient(Papp)determined in this study indicated that the permeability of SGs improved to the moderate class compared to the corresponding parent compounds,predicting a higher in vivo ***,the efflux ratio(ER)value demonstrated an active uptake of SSd and the three SGs,while a passive diffusion of SSa and SSb2.

关键词： Bupleuri Radix Saikosaponin Saikogenin UHPLC-MS/MS caco-2 cells Permeability

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