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| 1 | Oxidative stress-induced mitochondrial dysfunction in a normal colon epithelial cell line显示文摘AIM To determine how a normal human colon cell line reacts to microbial challenge as a way to study oxidative stress-induced responses associated with inflammatory bowel disease.METHODS Normal human colon epithelial cells(ATCC?CRL.1790?)were stimulated with either heat killed E.coli or heat killed murine cecal contents(HKC)and examined for several relevant biomarkers associated with inflammation and oxidative stress including cytokine production,mitochondrial autophagy and oxidant status.TNFα,IL-1βand IL-8 protein concentrations were measured within the supernatants.Fluorescent microscopy was performed to quantify the production of reactive oxygen species(ROS)using an oxidation responsive fluorogenic probe.Mitochondrial morphology and mitochondrial membrane potential was assessed by dual staining using COXIV antibody and a dye concentrating in active mitochondria.Mitochondrial ROS scavenger was used to determine the source of ROS in stimulated cells.Autophagy was detected by staining for the presence of autophagic vesicles.Positive controls for autophagy and ROS/RNS experiments were treated with rapamycin and chloroquine.Mitochondrial morphology,ROS production and autophagy microscopy experiments were analyzed using a custom acquisition and analysis microscopy software(Image J).RESULTS Exposing CRL.1790 cells to microbial challenge stimulated cells to produce several relevant biomarkers associated with inflammation and oxidative stress.Heat killed cecal contents treatment induced a 10-12fold increase in IL-8 production by CRL.1790 cells compared to unstimulated controls at 6 and 12 h(P<0.001).Heat killed E.coli stimulation resulted in a4-5 fold increase in IL-8 compared to the unstimulated control cells at each time point(P<0.001).Both heat killed E.coli and HKC stimulated robust ROS production at 6(P<0.001),and 12 h(P<0.01).Mitochondrial morphologic abnormalities were detected at 6 and12 h based on reduced mitochondrial circularity and decreased mitochondrial membrane potential,P<0.01.Microbial stimulation also induced significant autophagy at 6 and 12 h,P<0.01.Lastly,blocking mitochondrial ROS generation using mitochondrial specific ROS scavenger reversed microbial challenge induced mitochondrial morphologic abnormalities and autophagy.CONCLUSION The findings from this study suggest that CRL.1790cells may be a useful alternative to other colon cancer cell lines in studying the mechanisms of oxidative stress events associated with intestinal inflammatory disorders. | Nandakumar Packiriswamy Kari F Coulson Susan J Holcombe Lorraine M Sordillo | 2017 | World Journal of Gastroenterology2017，23，19： | 3 |
| 2 | Toward a model of strategic outsourcing显示文摘 | Holcomb T R Hitt M A | 2007 | Journal of Operations Management2007，25，2： | 1 |
| 3 | Why corals care about ocean acidification: Uncovering the mechanism显示文摘 | Cohen A L Holcomb M | 2009 | Oceanography2009，22，4： | 1 |
| 4 | Introducing X-machinemodels to verify PLC ladder diagrams显示文摘 | Chambers C Holcombe M Barnard J | 2001 | Computers in Industry2001，45，3： | 1 |
| 5 | Expression of Wnt ligands and Frizzled receptors in colonic mucosa and in colon carcinoma显示文摘 | Holcombe R F Marsh J L Waterman M L | 2002 | Mol Pathol2002，55，4： | 1 |
| 6 | Long-term effects of nutrient and CO2 emiehment on the temperate coral Astrangia poeulata ( Ellis and Solandtr, 1786 ) 显示文摘 | Holcomb M McCorkle D C Cohen A L | 2010 | Journal of experimental marine biology and ecology2010，386，12： | 1 |
| 7 | Properties influencing coolness to the touch of fabrics显示文摘 | Schneider A M Holcombe B V | 1991 | Textile Research Journal1991，61，： | 1 |
| 8 | Influence of prepa- ration conditions on the composition of type B carbonated hy- droxyapatite and on the localization of the carbonate ions显示文摘 | Vignoles M Bonel G Holcomb DW | 1988 | Calcif Tissue In/1988，43，1： | 1 |
| 9 | Determination of aflatoxins in food products by chromatography显示文摘 | Holcomb M Wilson DM Trucksess MW | 1992 | J Chromatogr1992，624，： | 1 |
| 10 | The role of nuclear factor kappaB in pancreatic cancer and the clinical applications of targeted therapy显示文摘 | HOLCOMB B Y1P-SCHNEIDER M SCHMIDT C M | 2008 | Pancreas2008，36，3： | 1 |
| 11 | Architecture of Entrepreneurial Learning: Ex ploring the Link among Heuristics, Knowledge, and Action显示文摘 | HOLCOMB T R IRELAND R D HOLMES R M | 2009 | Entrepreneurship Theory and Practice2009，33，1： | 1 |
| 12 | Preparation of Cr-doped Y3Al5O12 phosphors by heterogeneous precipitation methods and their luminescent properties显示文摘 | MATSUBARA I PARANTHAMAN M ALLISON S W CATES M R BESHEARS D L HOLCOMB D E | 2000 | Materials Research Bulletin2000，35，2： | 1 |
| 13 | The role of nuclear factor kappaB in pancreatic cancer and the clinical applications of targeted therapy 显示文摘 | Holcomb B Yip-Schneider M Schmidt CM | 2008 | Pancreas2008，36，3： | 1 |
| 14 | Coagulopathy: its pathophysiology and treatment in the injured patient显示文摘 | Tieu B H Holcomb J B Schreiber M A | 2007 | World J Surg2007，31，5： | 1 |
| 15 | The role of nuclear factor kappaB in pancreatic cancer and the clini- cal applications of targeted therapy显示文摘 | Holcomb B Yip-Schneider M Schmidt CM | 2008 | Pancreas2008，36，3： | 1 |
| 16 | The shippers'perspective : transportation and logistics trends and issues显示文摘 | HOLCOMB M C MANRODT K B | 2000 | Transporta- tion Journal2000，40，1： | 1 |
| 17 | Toward a model of strategic outsourcing显示文摘 | HOLCOMB R T HITT A M | 2007 | Journal of Operations Management2007，25，2： | 1 |
| 18 | Toward a model of strategic outsourcing显示文摘 | Holcomb T R Hitt M A | | 0，，25： | 1 |
| 19 | 显示文摘 | Chu Y H Martin L W Holcomb M B | 2007 | Materials Today2007，10，： | 1 |
| 20 | ophthahnopathie im- munoglobulin in patients with Graves' ophthalmopathy 显示文摘 | Atkinson S Holcombe M Kendall-Taylor P | 1984 | Lancet1984，324，8399： | 1 |
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