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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 | Improved outcome for haemodialysis patients treated with high flux memhranes显示文摘 | Woods HF Nandakumar M | 2000 | Nephrol Dial Transplant2000，15，1： | 1 |
| 3 | The state of proteome profiling in the fungal genus Aspergillus显示文摘 | KIM Y NANDAKUMAR M P MARTEN M R | 2008 | Brief Funct Genomic Proteomic2008，7，： | 1 |
| 4 | Improved outcome for haemodialysis patients treated with high flux membranes显示文摘 | Woods HF Nandakumar M | 2000 | Nephrol Diat Transplant2000，15，1： | 1 |
| 5 | Aberrant DNA hypermethylation patterns lead to transcriptional si- lencing of tumor suppressor genes in UVB-exposed skin and UVB-induced skin tumors of mice显示文摘 | Nandakumar V Vaid M Tollefsbol T O | 2011 | Carcinogene- sis2011，32，4： | 1 |
| 6 | Quantification of nisin in flow-injection immunoassay systems显示文摘 | Nandakumar R Nandakumar M P Mattiasson B | 2000 | Biosens Bioelectron2000，241，： | 1 |
| 7 | Mi-crobial glutaminase : biochemistry, molecular approachesand applications in the food industry 显示文摘 | Nandakumar R Yoshimune K Wakayama M | 2003 | Journal of Mo-lecular Catalysis B: Enzymatic2003，23，9： | 1 |
| 8 | Solubilization of trichloroacetic acid(TCA)precipitated microbial proteins via NaOH fortwo-dimensional gel electrophoresis 显示文摘 | Nandakumar M P | 2003 | J Proteome Res2003，2，1： | 1 |
| 9 | Solubiliza-tion of trichloroacetic acid(TCA)precipitated microbial proteins via NaOH for two-Dimensionalelectrophoresis显示文摘 | Nandakumar M P Jie Shen Babu Raman | | 0，，01： | 1 |
| 10 | Factors affecting the Agrobacterium-mediated transient transformation of the wetland monocot,Typha latifolia显示文摘 | Nandakumar R Chen L Rogers S M D | 2004 | Plant Cell Tiss Organ Cult2004，79，： | 1 |
| 11 | Microbial glutaminase: biochemistry, molecular approaches and applica- tions in the food industry 显示文摘 | Nandakumar K Yoshimune K Wakayama M | 2003 | Journal of Molecular Catalysis B: Enzymatic2003，，23： | 1 |
| 12 | (-) -Epigallocatechin- 3-gallate reactivates silenced tumor suppressor genes, Cipl/p21 and p16INK4a, by reducing DNA methylation and increasing histones aeetylation in human skin cancer cells 显示文摘 | Nandakumar V Vaid M Katiyar SK | 2011 | Carcinogenesis2011，32，： | 1 |
| 13 | Isocitrate lyase mediates broad antibiotic tolerance in Mycobacterium tuberculosis显示文摘 | NANDAKUMAR M NATHAN C RHEE K Y | 2014 | Nat Commun2014，5，5： | 1 |
| 14 | Solubilization of trichloroacetic acid (TCA) precipitated microbial proteins via NaOH for two-dimensional electrophoresis显示文摘 | Nandakumar M Shen J Raman B | 2003 | Journal of Proteome Research2003，2，1： | 1 |
| 15 | Theoretical correction of the Ouchiyama and Tanaka formula for predicting average porosity of packed beds consisting of nonuniform spheres 显示文摘 | Song M Chuang K T Nandakumar K | 1998 | Industrial and Engineering Chemistry Research1998，37，8： | 1 |
| 16 | Solubilization of trichloroacetic acid (TCA) precipitated microbial proteins via NaOH for two-dimensional electrophoresis显示文摘 | Nandakumar M P Shen J Raman B | | 0，，02： | 1 |
| 17 | Control relevant modeling of planer solid oxide fuel cell system 显示文摘 | MURSHED A M HUANG BIAO NANDAKUMAR K | 2007 | Journal of Power Sources2007，163，2： | 1 |
| 18 | Validation of kidney disease quality of life - short form: A cross sectional study of a dialysis- targeted heath meadure in Singapore显示文摘 | Veenan DJ Nandakumar M Jeremy FYL | 2010 | BMC Neph- rol2010，11，： | 1 |
| 19 | Use of a Micro-expanded Bed Containing Immobilised Lysozyme for Cell Disruption in Flow Injection Analysis 显示文摘 | Nandakumar M P Tocaj A Mattiasson B | 1999 | Bioseparation1999，8，： | 1 |
| 20 | Solubilization of trichloroaeetic acid (TCA) precipitated microbial proteins via NaOH for two-dimensional electrophoresis显示文摘 | NANDAKUMAR M P SHEN J RAMAN B | 2003 | Journal of Proteome Research2003，2，： | 1 |
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