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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 | Splenic Angiosarcoma显示文摘 | Emilia Mia Sordillo Peter P. Sordillo Steven I. Hajdu | 1995 | The American Journal of Surgical Pathology1995，，1： | 2 |
| 3 | Impact of oxidative stress on the health and immune function of dairy cattle显示文摘 | Lorraine M. Sordillo Stacey L. Aitken | 2008 | Veterinary Immunology and Immunopathology2008，，1： | 2 |
| 4 | Evaluation of antioxidant and proinflammatory gene expression in bovine mammary tissue during the periparturient period显示文摘 | S.L. Aitken E.L. Karcher P. Rezamand J.C. Gandy M.J. VandeHaar A.V. Capuco L.M. Sordillo | 2009 | Journal of Dairy Science2009，，2： | 2 |
| 5 | Multiple micro- bial exposures in the home may protect against asthma or allergy in childhood显示文摘 | Sordillo JE Hoflinan EB Celedon JC | 2010 | ClinExpAllergy2010，40，6： | 1 |
| 6 | Enhanced antigen-specific responses in bovine mammary glands following interleukin-2 administration显示文摘 | Pighetti G M Sordillo L M | 1995 | J Dairy Sci1995，78，： | 1 |
| 7 | Secretion composition during bovine mammary involution and the relationship with mastitis显示文摘 | Sordillo L Nickerson S Akers R | 1987 | Int J Biochem1987，19，12： | 1 |
| 8 | Immano-biology of the mammary gland显示文摘 | Sordillo L M | 1977 | Dairy Sci1977，80，： | 1 |
| 9 | Shifts in Thioredoxin Reductase Activity and Oxidant Status in Mononuclear Ceils Obtained from Transition Dairy Cattle显示文摘 | Sordillo L M O'Boyle N Gandy J C | 2007 | J Dairy Sci2007，90，3： | 1 |
| 10 | Factors affecting mammary gland immunity and mastitis susceptibility 显示文摘 | SORDILLO L M | 2005 | LivestockProduction Science2005，98，12： | 1 |
| 11 | Significance of metabolic stress,lipid mobilization,and inflammation on transition cow disorders显示文摘 | SORDILLO L M RAPHAEL W | 2013 | Veterinary Clinics of North America:Food Animal Practice2013，29，： | 1 |
| 12 | Immunobiology of the mammary gland 显示文摘 | Sordillo L M Shafer-Weaver K DeRosa D | 1997 | Dairy Sci1997，80，： | 1 |
| 13 | Effect of interferonγ on the production of tumor necrosis factor during acute Escherichia coli Mastitis 显示文摘 | SORDILLO L M PEEL J P | 1992 | J Dairy Sci1992，75，21： | 1 |
| 14 | Allergen exposure modifiesthe relation of sensitization to fraction of exhaled nitric oxide le-vels in children at risk for allergy and asthma显示文摘 | Sordillo JE Webb T Kwan D | 2011 | J Allergy Clin I-mmunol2011，127，5： | 1 |
| 15 | Impact of oxidative stress on the health and immune function of dairy cattle 显示文摘 | Sordillo L M Aitken S L | 2009 | Vet Immunol Immunop2009，128，： | 1 |
| 16 | Malignant peritoneal mesothelioma显示文摘 | Brenner J Sordillo PP Magi11 GB | 1981 | Am J Gastroenterol1981，75，： | 1 |
| 17 | Malignant schwannoma:clinical characteristics,survival,and response to therapy显示文摘 | Sordillo PP Helson L Hajdu SI | 1981 | Cancer1981，47，10： | 1 |
| 18 | Controlling acute Escherichia coli mastitis during the periparturient period with recombinant bovine interferon - gamma显示文摘 | SORDILLO L M BABINK L A | 2000 | J Gen Virol2000，81，： | 1 |
| 19 | Mammary Gland Immunity and Mastitis Susceptibility显示文摘 | Lorraine M Sordillo Katie L S | 2002 | Journal of Mammary Gland Biology and Neoplasia2002，，7： | 1 |
| 20 | Mammary gland immunity and mastitis susceptibility 显示文摘 | Sordillo L M Streicher K L | 2002 | Journal of Mammary Gland Biology and Neoplasia2002，7，： | 1 |
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