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Aim To investigate the effect of Sijunzi Decoction on mRNA and protein expression related to growth and cell cycle in polyamine/HuR signaling pathway during small intestinal epithelial cell (IEC-6) proliferation, and to explore its mechanism on intestinal mucosal injury repair. Methods Sijunzi Decoction-containing serum (SJZD) was prepared from SD rats, the expression of HuR protein in cytoplasm and nucleus was analyzed by immunofluorescence and Western blot, the mRNA level of activating transcription factor-2 (A T F - 2), JunD and cyclin dependent kinase 4 (CDK4) were determined by real-time fluorescent quantitative PCR (RT-PCR), Western blot was used to detect protein level of HuR, ATF-2, JunD and CDK4, and flow cytometry was applied to analyse cell cycle distribution. Results Compared with the control group, the mRNA and protein expression of ATF-2 and JunD decreased, while the expression of Cdk4 mRNA and protein increased in SZJD group, and the proportion of G
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Objective: To compare the anti-proliferative effect of sodium thiosulfate on human colorectal cancer cells (HT-29) and normal small intestine cells (IEC6). Methods: Cells (HT-29 and IEC6) were treated with different concentrations of sodium thiosulfate ranging from 0.5 mM to 80 mM for 24 h. Cell viability was measured via crystal violet and MTT assays. HT-29 cells were further treated in the presence and absence of mitochondrial electron transport chain (ETC) inhibitors, K
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Objective: To investigate the protection mechanism of Fushen Granule on LPS-induced intestinal epithelial cell (IEC-6) barrier injury. Methods: IEC-6 were divided into control group, model group, Fushen granule group, and p38MAPK inhibitor group. In addition to the control group, 1 μg/mL LPS was added to the other groups, 10% drug-containing serum and 10 μmol/L SB203580 were respectively added to the administration group. The apoptosis of IEC-6 were detected by flow cytometry; The permeability and transmembrane resistance of IEC-6 barrier were detected by Transwell assay; The mRNA levels of ICAM-1 and IL-1β were assessed by Real-time PCR; The expressions of occludin, ZO-1, p38MAPK, and DUSP1 were detected by western blotting. Results: Compared with the control group, the apoptosis of IEC-6 were significantly increased in model group (P < 0.01), the levels of ICAM-1 and IL-1β were significantly increased (P < 0.01), the occludin and ZO-1 were significantly decreased (P < 0.01), the intestinal permeability was significantly increased (P < 0.05), the mRNA levels of ICAM-1 and IL-1β in IEC-6 were significantly inhibited, the permeability of intestinal mucosal barrier was reduced (P < 0.05), the expressions of DUSP1, occludin and ZO-1were increased (P < 0.05, 0.01), the expression of p38MAPK was inhibited (P < 0.01), which showed a similar effect with p38MAPK inhibitor. Conclusion: Fushen Granule could regulate DUSP1 which inhibits the activation of p38MAPK signaling pathway, up-regulate Occludin and ZO-1, improve the tight junction of IEC-6, thereby improving the cell barrier function damage caused by LPS.
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Objective: To compare the anti-proliferative effect of sodium thiosulfate on human colorectal cancer cells (HT-29) and normal small intestine cells (IEC6). Methods: Cells (HT-29 and IEC6) were treated with different concentrations of sodium thiosulfate ranging from 0.5 mM to 80 mM for 24 h. Cell viability was measured via crystal violet and MTT assays. HT-29 cells were further treated in the presence and absence of mitochondrial electron transport chain (ETC) inhibitors, KATP channel opener and closer and H2S inhibitors for 24 h followed by sodium thiosulfate in order to study their respective roles in the anti-proliferative activity of sodium thiosulfate. Results: The IC50 values of sodium thiosulfate on HT-29 cells were 40.93 mM and 42.45 mM by crystal violet and MTT assay whereas, in the case of IEC6 cells, the values were 45.17 mM and 47.22 mM. The inhibition of endogenous H2S enzymes and KATP channel induced no change in the anti-proliferative capacity of sodium thiosulfate. However, the anti-proliferative activity of sodium thiosulfate was enhanced in the presence of mitochondrial ETC inhibitors. Conclusions: HT-29 cell growth is effectively attenuated by sodium thiosulfate and the anti-proliferative activity of sodium thiosulfate is enhanced in the presence of mitochondrial ETC inhibitors.
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OBJECTIVE： To compare low-polarity volatile constituents in supercritical CO2 extract from the roots and stem of Ilex asprella and its effects on the proliferation of IEC-6 in vitro， and to provide reference for making full use of wild resources of I. asprella and expanding its medicinal parts. METHODS： The low-polarity volatile constituents were extracted from the root and stem of I. asprella with supercritical fluid CO2 extraction（SFE-CO2）. The chemical constituents were analyzed by GC-MS. IEC-6 cells were treated with different concentrations of supercritical CO2 extracts （0， 1， 5， 10， 20， 40， 60， 80， 100 μg/mL） from roots or stems of I. asprella. MTT assay was used to detect the relative viability， and cell proliferation curve was drawn and EC50 of each extract were calculated. RESULTS： Sixty-two and forty-six low-polarity volatile constituents were identified from supercritical CO2 extract in the roots and stem of I. asprella with GC-MS； there were 24 common constituents totally， mainly including pelargonic acid（14.18％ and 6.14％），octanoic acid（10.59％ and 4.35％），hexanoic acid（8.63％ and 10.86％），paeonol（7.79％ and 6.00％），2-methyl-3-phenyl-propanal（6.3％ and 0.58％），acetic acid（1.72％ and 33.77％） in root and stem， respectively. The results of cell culture in vitro showed that when the concentration of supercritical CO2 extract from the roots and stems of I. asprella was lower （≤60 μg/mL）， it could significantly promote the proliferation of IEC-6 cells and their EC50 were 16.35， 20.20 μg/mL， respectively； when the concentration of the extract was higher （≥80 μg/mL）， it showed cytotoxicity and inhibited the proliferation of IEC-6 cells. CONCLUSIONS： There are similar species of volatile constituents in roots and stems of I. asprella and similar in vitro bioactivity of the supercritical CO2 extracts to IEC-6 cells. The short-chain fatty acids may be the active ingredient to promote cell proliferation， while paeonol may be the cytotoxic active ingredient.
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Astragalus membranaceus (Radix Astragali, RA) and Atractylodes macrocephala (Rhizoma Atractylodis Macrocephalae, RAM) are often used to treat gastrointestinal diseases. In the present study, we determined the effects of polysaccharides extracts from these two herbs on IEC-6 cell migration and explored the potential underlying mechanisms. A migration model with IEC-6 cells was induced using a single-edged razor blade along the diameter of cell layers in six-well polystyrene plates. The cells were grown in control media or media containing spermidine (5 μmol·L, SPD), alpha-difluoromethylornithine (2.5 mmol·L, DFMO), 4-Aminopyridine (40 μmol·L, 4-AP), the polysaccharide extracts of RA or RAM (50, 100, or 200 mg·L), DFMO plus SPD, or DFMO plus polysaccharide extracts of RA or RAM for 12 or 24 h. Next, cytosolic free Ca ([Ca]) was measured using laser confocal microscopy, and cellular polyamine content was quantified with HPLC. Kv1.1 mRNA expression was assessed using RT-qPCR and Kv1.1 and RhoA protein expressions were measured with Western blotting analysis. A cell migration assay was carried out using Image-Pro Plus software. In addition, GC-MS was introduced to analyze the monosaccharide composition of both polysaccharide extracts. The resutls showed that treatment with polysaccharide extracts of RA or RAM significantly increased cellular polyamine content, elevated [Ca] and accelerated migration of IEC-6 cells, compared with the controls (P < 0.01). Polysaccharide extracts not only reversed the inhibitory effects of DFMO on cellular polyamine content and [Ca], but also restored IEC-6 cell migration to control level (P < 0.01 or < 0.05). Kv1.1 mRNA and protein expressions were increased (P < 0.05) after polysaccharide extract treatment in polyamine-deficient IEC-6 cells and RhoA protein expression was increased. Molar ratios of D-ribose, D-arabinose, L-rhamnose, D-mannose, D-glucose, and D-galactose was 1.0 : 14.1 : 0.3 : 19.9 : 181.3 : 6.3 in RA and 1.0 : 4.3 : 0.1 : 5.7 : 2.8 : 2.2 in RAM. In conclusion, treatment with RA and RAM polysaccharide extracts stimulated migration of intestinal epithelial cells via a polyamine-Kv1.1 channel activated signaling pathway, which facilitated intestinal injury healing.
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Animals , Rats , Astragalus propinquus , Chemistry , Atractylodes , Chemistry , Cell Line , Cell Movement , Drugs, Chinese Herbal , Chemistry , Pharmacology , Epithelial Cells , Cell Biology , Metabolism , Intestines , Cell Biology , Genetics , Metabolism , Polyamines , Metabolism , Polysaccharides , Chemistry , Pharmacology , Rhizome , Chemistry , Signal Transduction , rhoA GTP-Binding Protein , MetabolismABSTRACT
Aim To observe the effect of epidermal growth factor (EGF) on cell growth under different culture conditions using the real-time cell analyzer and EGF as a tool medicine to promote cell growth,and to provide reference for establishing pharmacokinetic model of IEC-6 cell growth (proliferation).Methods IEC-6 cell was inoculated on E-Plate 16 plate at a density of 1 × 104 cell/well and cultured in DMEM with 10％ serum for 24 h,then replaced by serum-free DMEM culture (serum starvation)for 20 h,then the effects of different culture conditions on cell growth as well as EGF efficacy were observed.Results ① When the serum concentration was 10％,the cell growth index of EGF group(1,10,100 μg · L-1) after drug administration 24 h,48 h and 72 h was P ＞ 0.05 compared with the blank group,suggesting that 10％ serum culture could not reflect the efficacy of EGF.② When the serum concentration was 0％,EGF (1,10 μg · L-1) improved cell growth inhibition caused by serum-free cultivation,but could not recover it to normal level (the EGF group after drug administration 24 h,48 h and 72 h was P ＜0.01 compared with 5％ serum),which suggested that serum-free culture could not reflect the EGF efficacy.③ 0％,0.5％,1％ serum had different effects on cell growth,of which 0.5％ serum could neither have obvious inhibition on cell growth,nor reflect the EGF effect due to promoting cell growth for a long time.④When the serum concentration was 0.5 ％,the cell growth index of EGF groups after drug administration 24h,48h and 72h was P ＜0.01 compared with the blank group,suggesting that 0.5％ serum culture could better reflect EGF efficacy.⑤The efficacy of EGF (10 μg· L-1) in promoting cell growth was confirmed by repeated validation of 0.5 ％ serum.Condusions A reference scheme of the IEC-6 cell growth (proliferative) pharmacological experimental model is established in the real-time cell analyzer:cells are cultured in DMEM with 10％ serum for 24h,then in serum-free DMEM (serum starvation) for 20h,then after the adding of reagent,cells are cultured in DMEM with 0.5％ serum for 48-72 h to observe its effect on cell growth (proliferation).
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BACKGROUND: The aggravating factors still remained unclear in inflammatory bowel disease (IBD). Despite many different therapeutic approaches, many patients do not respond to the therapy. The anti-inflammatory effect of insulin-like growth factor-binding protein-3 (IGFBP-3) was suggested because of its capability of nuclear factor-κB (NF-κB) signaling inhibition. Therefore, we hypothesized that the up-regulation of IGFBP-3 would inhibit an inflammatory process. METHODS: Lipopolysaccharides (LPS) treated intestinal epithelial cell 6 (IEC-6) and dextran sodium sulfate (DSS) induced colitis mice were used as colitis models. Exogenous IGFBP-3 expression was accomplished using the adenoviral vector system expressing IGFBP-3 (Ad/IGFBP-3). The inflammatory responses and relevant cellular responses in IEC-6 cells influenced by IGFBP-3 expression were evaluated by western blotting, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and reactive oxygen species (ROS) measurement. The severity of colitis was evaluated with the colon tissues of DSS-induced mouse model. RESULTS: We found that the IGFBP-3 expression reduced the production of inflammatory cytokines (cyclooxygenase-2, interleukin-1β, tumor necrosis factor-α) and ROS formation. IGFBP-3 expression also induced cell viability and inhibited NF-κB activation. In line with this data, the severity of DSS-induced mouse colitis was greatly ameliorated by the treatment of IGFBP-3 expressing adenoviral particles characterized with less weight loss and preserved colon length compared with the mice treated with DSS alone. The histopathology of the colon showed the reducing signs of colitis in Ad/IGFBP-3 treated DSS-mice group. CONCLUSION: Therefore, our data suggest that Ad/IGFBP-3 up-regulation reduces colonic inflammatory response as a novel therapeutic protocol for IBD.
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Animals , Humans , Mice , Blotting, Western , Cell Survival , Colitis , Colon , Cytokines , Dextrans , Epithelial Cells , Inflammatory Bowel Diseases , Insulin-Like Growth Factor Binding Protein 3 , Lipopolysaccharides , Necrosis , Reactive Oxygen Species , Sodium , Up-Regulation , Weight LossABSTRACT
Astragalus membranaceus (Radix Astragali, RA) and Atractylodes macrocephala (Rhizoma Atractylodis Macrocephalae, RAM) are often used to treat gastrointestinal diseases. In the present study, we determined the effects of polysaccharides extracts from these two herbs on IEC-6 cell migration and explored the potential underlying mechanisms. A migration model with IEC-6 cells was induced using a single-edged razor blade along the diameter of cell layers in six-well polystyrene plates. The cells were grown in control media or media containing spermidine (5 μmol·L, SPD), alpha-difluoromethylornithine (2.5 mmol·L, DFMO), 4-Aminopyridine (40 μmol·L, 4-AP), the polysaccharide extracts of RA or RAM (50, 100, or 200 mg·L), DFMO plus SPD, or DFMO plus polysaccharide extracts of RA or RAM for 12 or 24 h. Next, cytosolic free Ca ([Ca]) was measured using laser confocal microscopy, and cellular polyamine content was quantified with HPLC. Kv1.1 mRNA expression was assessed using RT-qPCR and Kv1.1 and RhoA protein expressions were measured with Western blotting analysis. A cell migration assay was carried out using Image-Pro Plus software. In addition, GC-MS was introduced to analyze the monosaccharide composition of both polysaccharide extracts. The resutls showed that treatment with polysaccharide extracts of RA or RAM significantly increased cellular polyamine content, elevated [Ca] and accelerated migration of IEC-6 cells, compared with the controls (P < 0.01). Polysaccharide extracts not only reversed the inhibitory effects of DFMO on cellular polyamine content and [Ca], but also restored IEC-6 cell migration to control level (P < 0.01 or < 0.05). Kv1.1 mRNA and protein expressions were increased (P < 0.05) after polysaccharide extract treatment in polyamine-deficient IEC-6 cells and RhoA protein expression was increased. Molar ratios of D-ribose, D-arabinose, L-rhamnose, D-mannose, D-glucose, and D-galactose was 1.0 : 14.1 : 0.3 : 19.9 : 181.3 : 6.3 in RA and 1.0 : 4.3 : 0.1 : 5.7 : 2.8 : 2.2 in RAM. In conclusion, treatment with RA and RAM polysaccharide extracts stimulated migration of intestinal epithelial cells via a polyamine-Kv1.1 channel activated signaling pathway, which facilitated intestinal injury healing.
Subject(s)
Animals , Rats , Astragalus propinquus , Chemistry , Atractylodes , Chemistry , Cell Line , Cell Movement , Drugs, Chinese Herbal , Chemistry , Pharmacology , Epithelial Cells , Cell Biology , Metabolism , Intestines , Cell Biology , Genetics , Metabolism , Polyamines , Metabolism , Polysaccharides , Chemistry , Pharmacology , Rhizome , Chemistry , Signal Transduction , rhoA GTP-Binding Protein , MetabolismABSTRACT
AIM: To observe the effects of miR-542-5p on the proliferation of rat small intestine crypt epithe-lial IEC-6 cells induced by sphingosine-1-phosphate (S1P).METHODS: Two IEC-6 cell lines (SphK1-IEC-C1 and SphK1-IEC-C2) were established, which expressed sphingosine kinase-1 (SphK1) stably.Radioactive tracer was used to detect SphK1 activity and S1P secretion.The cell proliferation was observed by cell counting and described by drawing growth curve, and the cell cycle analysis was carried out by flow cytometry.The level of miR-542-5p was evaluated by RT-qPCR.RESULTS: Compared with control vector cells without SphK1 cDNA, both SphK1-IEC-C1 and SphK1-IEC-C2 cell lines showed that Sphk1 was elevated, both intracellular and extracellular S1P increased dramatically, the rate of cell growth was faster, the percentage of the cells in S phase increased, and miR-542-5p expression decreased.S1P (0.5~10 μmol/L) led to the decrease in miR-542-5p expression.On the contrary, SphK1 silencing resulted in the increase in miR-542-5p expression in the IEC-6 cells.The miR-542-5p was elevated in SphK1-IEC-C1 cells and SphK1-IEC-C2 cells, which caused the decrease in the percentage of the cells in S phase.The cell growth rate in the above-mentioned 2 cell lines decreased compared with negative control group.CONCLUSION: In IEC-6 cells, S1P promotes proliferation by inhibiting miR-542-5p expression, which induces the cell cycle transferring from G1 phase to S phase.
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[ ABSTRACT] AIM:To investigate the role of protease activated receptor-2 ( PAR-2 ) in the process of tryptase mediated IEC-6 cell injury.METHODS:The rat intestinal epithelial cell line IEC-6 was treated with tryptase at different concentrations (1 μg/L, 10 μg/L, 100μg/L and 1 000μg/L) in the presence or absence of PAR-2 antagonist FSLLRY-NH2 for 12 h respectively.The cell survival rate was detected by MTT assay.The protein levels of PAR-2 and cleaved-caspase 3 were determined by Western blotting.The LDH activity was also measured.RESULTS:Compared with control group, the cell survival rates were significantly decreased in 100 μg/L and 1 000 μg/L tryptase treated groups, the LDH activities were significantly increased in 10 μg/L to 1 000 μg/L tryptase treated groups, and the protein levels of PAR-2 and cleaved caspase 3 were significantly increased in 100μg/L and 1 000μg/L tryptase treated groups (P<0.05).Com-pared with 1 000 μg/L tryptase treated group, the LDH activity and cleaved caspase 3 protein level were dramatically de-creased while the survival rate was significantly increased in the presence of PAR-2 antagonist FSLLRY-NH2 (P<0.05). CONCLUSION:Tryptase induces IEC-6 cell injury in a dose-dependent manner by activating PAR-2.
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Objective: To observe the effect of flavonoid from Glycyrrhizae Radix (FGR) on the migration of small intestinal epithelial cells (IEC-6) and intracellular polyamines content. Methods: The cell migration model was constructed by scratch method. Under the intervention of α-difluoromethylornithine (DFMO), the specific inhibitor of polyamine synthesis, or 4-aminopyridine (4-AP), the specific inhibitor of potassium channel, the effect of FGR on cell migration was observed. The intracellular polyamine content being cultured for 12 or 24 h in normal and DFMO intervention condition was detected by precolumn derivatization-HPLC. Results: FGR could reverse the inhibitory effects of DFMO and 4-AP on cell migration, increase the intracellular content of polyamines after 12 or 24 h treatment, and reserve the decrease of polyamine content caused by DFMO. Conclusion: FGR could affect the polyamine-dependent signaling pathway, which may be related to the mechanisms for FGR promoting cell migration.
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Objective To observe the effects of quercetin on PGE 2 release and cyclooxygenase activity in IEC 6 cells stimulated by the serum of burned rats. Methods The PGE 2 levels were measured by radioimmunoassay and the changes of cyclooxygenase activity were determined by substrate fluorescence analysis. Results Stimulated by the serum of burned rats, the PGE 2 level increased first and then obviously decreased. Quercetin at 0.1 and 1.0 ?mol/L enhanced the PGE 2 level after the stimulation of IEC 6 cells for 24 h. There was no obvious change in total cyclooxygenase activity, but the activity ratio of COX 1 to COX 2 significantly decreased. Conclusion Quercetin might influence the release of cyclooxygenase metabolite PGE 2 through the regulation of COXs activity in IEC 6 cells.
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Objective To study the effect of Bifidobacterium spent culture supernatant (SCS) on adhesion of Pseudomonas aeruginosa to intestinal epithelial cells. Methods The intestinal epithelial cells were cultured,then divided into three groups: control,Pseudomonas aeruginosa adhesion group,SCS and Pseudomonas aeruginosa adhesion group. The effects of SCS on cell viability,the number of adhering and invasive bacteria were evaluated with MTT assay and lysis-counting assay in 1,3,6 h. Results At 3 h after SCS treatment,the amount of Pseudomonas aeruginosa decreased significantly,and the number of adhering and invasive bacteria decreased by 71% and 87% respectively. Conclusion SCS could protect the intestinal epithelial cells by inhibiting the adhesion and invasion of Pseudomonas aeruginosa.
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Objective:Glutamine is the main oxidative fuel of the enterocyte which enters the enterocyte primarily via amino acid transporters.The aim of the test was to study the distributions and functions of glutamine transporters in IEC-6 cell line.Methods:The rat intestinal epithelial cell line(IEC-6) was incubated in vitro.The mRNA expression of different glutamine transporters,protein expression of system ASCT2,and the [3H]-L-glutamine uptake were measured.Results:The mRNA of system ASCT2,SN1,ATA1,LAT1,LAT2 was expressed and the protein expression of ASCT2 was also validated in IEC-6.In Na+-containing buffer,the velocity of Na+-dependent glutamine uptake was(164.07?37.94) fmol/(mg protein?10min).In Na+-free buffer,the velocity of glutamine uptake was(58.71?10.51)fmol/(mg protein?10min).With the saturate dosage of MeAIB,the velocity of glutamine uptake was(81.02 ?19.59) fmol/(mg protein?10min).Conclusion:There may be five kinds of glutamine transporters(ASCT2,SN1,ATA1,LAT1,and LAT2) in IEC-6 cell.The Na+-dependent transporter was the major contributor(64.22%) to glutamine total uptake in IEC-6.The contributions of system A and the remainder were 50.62% and 13.60%,respectively.The Na+-independent transporter was the lesser contributor(35.78%).
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Aim To investigate the protective effect of RTP1,one of the polysaccharide isolated from Rheum tanguticum,on H2O2 induced apoptosis in IEC-6 cells and its possible mechanism.Methods H2O2(100 mmol?L~-1)was used to induce IEC-6 cell apoptosis.Different doses(10,30,100 mg?L~-1)of RTP1 were administrated before H2O2 was added into IEC-6 cell culture.Cell viability was observed by MTT assay.Reactive oxygen species were measured with laser scanning confocal microscopy(LSCM).DNA content and percentage of apoptosis were assayed by DNA agarose gel electrophoresis,acridine orange staining and flow cytometry.The activation of Caspase-3 was detected with Western blot analysis.Results Following treatment with H2O2 for 2 h,H2O2 induced a significant decrease in cell viability,while DNA ladder was observed and apoptosis percentage was as high as 31.3%.Accumulation of intracellular ROS and increase in Caspase-3 activity were also detected.Pretreatment with RTP1 for 24 h exhibited cytoprotective effects in a dose-dependent manner.RTP1 obviously enhanced cell viability,reduced formation of DNA ladder and significantly reduced the number of cells labeled with Annexin V.The percentage of apoptosis/necrosis cells was markedly decreased to 24.4% and 21.5%,respectively.LSCM showed that RTP1 attenuated the accumulation of ROS.The significant decrease in Caspase-3 activity was detected.Conclusion RTP1 has cytoprotective capacity to antagonize H2O2-induced IEC-6 cell apoptosis and injury,and this effect may be related to decrease ROS and inhibit Caspase-3 activity
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PURPOSE: We investigated compounds from food sources given to children that may induce the differentiation of small intestinal epithelial cells in order to signal pathways that induce the prolif eration and differentiation of small intestinal epithelial cells. METHODS: We analyzed small intestinal epithelial cell differentiation using in vitro IEC-6 cells model. The growth curve of IEC-6 cells was obtained by standard MTT assay. Alkaline phospha tase(ALP) activities were determined using the paranitrophenol colorimetric assay for the differ entiation of IEC-6 cells. We did ALP and Brdu double-staining of cultured IEC-6 cells to distin guish between differentiation and proliferation, and investigated compounds' potential for inducing differentiation of small intestinal epithelial cells and protein kinase signal pathway. RESULTS: The calcium ion was essential for the differentiation of IEC-6 cells. Retinol and retinoic acid induced the differentiation of IEC-6 cells. beta-LG stabilized and increased cell permeation of retinoic acid. IEC-6 cells showed 3 or 4 times more ALP activity with co-treatment of retinoic acid and beta-LG. BSA and OVA accelerated differentiation of IEC-6 cells in a similiar fashion to beta -LG. But, pepton and casein didn't. Heat destruction of beta-LG, BSA and OVA lead to loss in the ability of these compounds to induce cellular differentiation. The PKA signal pathway involved differentiation of IEC-6 cells. IEC-6 cells proliferation increased by the activation of PKC signal pathway and decreased differentiation by PKC signal pathway. CONCLUSION: Our results confirm that signal pathways are related to the proliferation and differ entiation of small intestinal epithelial cells and various compounds from food sources of childhood, such as beta-LG, BSA, OVA, and retinoic acid. These compounds appear to induce differentiation of small intestinal epithelial cells and may play a role in stimulating regeneration of epithelial cells after small intestinal mucosal injury.
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Child , Humans , Bromodeoxyuridine , Calcium , Caseins , Epithelial Cells , Hot Temperature , Ovum , Protein Kinases , Regeneration , Signal Transduction , Tretinoin , Vitamin AABSTRACT
Object To observe the effects of Astragali Injection (Hi) and extracts from Atractylodes macrocephala Koidz. (B1, B4, B9, B13) on the migration of rat small intestinal crypt like cells line (IEC 6). Methods IEC 6 cells were inoculated in 6 well microplates and injured in 72 h, and added with Hi, B1, B4, B9, B13. Then D PBS to negative control in the same amount and EGF to positived control, the cell migration was observed in 24, 48 and 72 h after treatment. Results B4?B9?B13 and EGF significantly promoted the migration of wounded IEC 6 cells (P
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Objective To study the effects of Chinese herb extracts on cell proliferation and glucose absorption in intestinal epithelial cell line(IEC-6 cells) of rats.Methods The extracts of four Chinese herbs,including Herba Agastaches(HA),Rhizoma Atractylodis(RA), Cortex Phellodendri(CP),and Gypsum Fibrosum(GF),were made.Their suitable concentration on the cell proliferation in IEC-6 cells was determined by MTT method,and glucose absorption and the activity of Na+,K+-ATPase in IEC-6 cells were assayed.A method of real time PCR was applied to the determination of SGLT1 and GLUT2 mRNA expression in the cells.Results Chinese herb extracts treatment altered the cell proliferation in a dose-dependent manner.Moreover,RA volatile oil(50 ?g/mL) and CP alkaloid(10 ?g/mL) treatment increased glucose absorption and the activity of Na+,K+-ATPase genes(P0.05).Conclusion The three extracts treatment,including RA volatile oil,CP aklaloid,and HA volatile oil,could increase glucose absorption and the expression of glucose transport carrier genes,but their regulative mechanism are not totally the same.
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Objective To explore the activation of PI3k/Akt pathway of serum deprived IEC-6 cells by the serum of rats exposed to single radiation,burn or combined injury.Methods The IEC-6 cells were cultured in serum deprived media for 24 h,and stimulated by the serum of rats exposed to single radiation(~(60)Co ? ray at dose of 9 Gy),single burn(exposure to 5 kW tungsten-halogen light till whole body Ⅲ degree burn) or combined injury(burn first and radiation),and the cells stimulated by the serum from the normal rats and serum starved cells served as the control group.The total proteins of different group cells were extracted and the levels of phosphorylation of Akt were tested by Western blotting.The differentially expressed low mass proteins in the serums were detected by SELDI proteinchip technology,and primarily analyzed by related software as well as bioinformatic methods.Results The level of phosphorylation of Akt in the IEC-6 cells stimulated by serum from rats exposed to single radiation,single burn or combined injury was higher than in the cells stimulated by the serum from normal rats,in which the burn serum caused the highest level.As compared to burn rat serum,the serum of radiation and combined injury had 13 and 6 differentially expressed protein peaks respectively.Conclusion All the serums from rats exposed to different kinds of damage agents could activate the PI3K/Akt pathway of IEC-6 cells efficiently.The special components of burnt rat serum may contribute to the highest effect on the phosphorylation of Akt.