)| PMID |
12614931 ( ) |
|---|---|
| Title | Mitochondrial dysfunction and death in motor neurons exposed to the glutathione-depleting agent ethacrynic acid. |
| Abstract | This study investigated the mechanisms of toxicity of glutathione (GSH) depletion in one cell type, the motor neuron. Ethacrynic acid (EA) (100 microM) was added to immortalized mouse motor neurons (NSC-34) to deplete both cytosolic and mitochondrial glutathione rapidly. This caused a drop in GSH to 25% of the initial level in 1 h and complete loss in 4 h. This effect was accompanied by enhanced generation of reactive oxygen species (ROS) with a peak after 2 h of exposure, and by signs of mitochondrial dysfunction such as a decrease in 3-(4,5-dimethyl-2-thiazoyl)-2,5-diphenyltetrazolium bromide (MTT) (30% less after 4 h). The increase in ROS and the MTT reduction were both EA concentration-dependent. Expression of heme oxygenase-1 (HO-1), a marker of oxidative stress, also increased. The mitochondrial damage was monitored by measuring the mitochondrial membrane potential (MMP) from the uptake of rhodamine 123 into mitochondria. MMP dropped (20%) after only 1 h exposure to EA, and slowly continued to decline until 3 h, with a steep drop at 5 h (50% decrease), i.e. after the complete GSH loss. Quantification of DNA fragmentation by the TUNEL technique showed that the proportion of cells with fragmented nuclei rose from 10% after 5 h EA exposure to about 65% at 18 h. These results indicate that EA-induced GSH depletion rapidly impairs the mitochondrial function of motor neurons, and this precedes cell death. This experimental model of oxidative toxicity could be useful to study mechanisms of diseases like spinal cord injury (SCI) and amyotrophic lateral sclerosis (ALS), where motor neurons are the vulnerable population and oxidative stress has a pathogenic role. Milan, Italy. |
NOTE: Color highlight is limited to the abstract and SciMiner text-mining mode. If you see much more identified targets below from "Targets by SciMiner Summary" and "Targets by SciMiner Full list", they may have been identified from the full text.
Targets by SciMiner Summary
| HUGO ID | Symbol | Target Name | #Occur | ActualStr |
|---|---|---|---|---|
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | 24 | HO-1 | heme oxygenase 1 | ho 1 | |
| 1516 | CAT | catalase | 4 | catalase | |
| 670 | RHOD | ras homolog gene family, member D | 4 | Rho | |
| 6871 | MAPK1 | mitogen-activated protein kinase 1 | 2 | MAPK | p38 | |
| 2983 | DNTT | deoxynucleotidyltransferase, terminal | 2 | TdT | |
| 6876 | MAPK14 | mitogen-activated protein kinase 14 | 1 | p38 mitogen activated protein kinase | |
| 10940 | SLC1A2 | solute carrier family 1 (glial high affinity glutamate transporter), member 2 | 1 | EAAT2 | |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | 1 | superoxide dismutase | |
Targets by SciMiner Full list
| HUGO ID | Symbol | Name | ActualStr | Score | FlankingText |
|---|---|---|---|---|---|
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | HO-1 | 3.4 | Expression of heme oxygenase-1 (HO-1), HO-1 a marker of oxidative stress also increased |
| 10940 | SLC1A2 | solute carrier family 1 (glial high affinity glutamate transporter), member 2 | EAAT2 | 1.5 | receptor subunit and a much greater perisomatic expression of the EAAT2 protein the glial glutamate transporter which is particularly vulnerable to |
| 670 | RHOD | ras homolog gene family, member D | Rho | 0.0 | measured by uptake of the lipophilic cation rhodamine 123 (Rho Rho 123 Fluka Chemie CH into mitochondria |
| 670 | RHOD | ras homolog gene family, member D | Rho | 0.0 | The cells were then resuspended in 1 ml of Rho 123 (10 10 _amp_#x3bc;g/ml) _amp_#x3bc g ml for 30 min |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | HO-1 | 3.4 | HO-1 mRNA |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | HO-1 | 3.4 | The heme oxygenase-1 (HO-1) HO-1 mRNA level was measured by Northern Blot analysis as previously |
| 2983 | DNTT | deoxynucleotidyltransferase, terminal | TdT | 0.3 | by the TUNEL technique which uses terminal deoxynucleotidyl transferase (TdT) TdT to catalyse the binding of FITC-conjugated dUTP to DNA strand |
| 2983 | DNTT | deoxynucleotidyltransferase, terminal | TdT | 0.3 | mixture (Roche Roche Molecular Biochemicals Monza Italy containing dUTP-FITC and TdT and incubated for 60 min at 37 _amp_#xb0 C in |
| 670 | RHOD | ras homolog gene family, member D | Rho | 0.0 | NSC-34 motor neurons by selective uptake of the lipophilic cation Rho 123 into mitochondria ( Fig 3 |
| 670 | RHOD | ras homolog gene family, member D | Rho | 0.0 | The histograms of relative MMP measured by fluorescent emission from Rho 123 taken up by mitochondria in control and EA-exposed NSC-34 |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | HO-1 | 3.4 | marker of oxidative stress we also measured the induction of HO-1 mRNA |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | HO-1 | 3.4 | Exposure to 100 _amp_#x3bc M EA promptly increased HO-1 mRNA with a massive rise 3 h after treatment (respectively |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | HO-1 | 3.4 | GSH depletion by EA induced expression of HO-1 which has been reported to be protective in different models |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | HO-1 | 3.4 | The end products of HO-1 enzymatic activity potentially act as antioxidants 45 and can thus |
| 6871 | MAPK1 | mitogen-activated protein kinase 1 | p38 | 1.7 | manner as an anti-apoptotic messenger possibly through activation of the p38 mitogen-activated protein kinase (MAPK) MAPK signal transduction pathway 46 |
| 6871 | MAPK1 | mitogen-activated protein kinase 1 | MAPK | 1.7 | possibly through activation of the p38 mitogen-activated protein kinase (MAPK) MAPK signal transduction pathway 46 |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | HO-1 | 3.4 | However the role of HO-1 in our model requires further investigation since proapoptotic signals prevailed |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | HO-1 | 3.4 | Interestingly increased immunoreactivity for HO-1 and p38MAPK were detected in human and mouse ALS 47 |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | HO-1 | 3.4 | The heme oxygenase-1 (HO-1) HO-1 mRNA signal is shown |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | ho 1 | 1.0 | expression of heme oxygenase 1 ho 1 a marker of oxidative stress also increased. |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | heme oxygenase 1 | 1.0 | expression of heme oxygenase 1 ho 1 a marker of oxidative stress also increased. |
| 1516 | CAT | catalase | catalase | 1.0 | ecause of its high level of polyunsaturated fatty acids as substrates for lipid peroxidation high rate of oxygen consumption and low or moderate levels of the antioxidant enzymes superoxide dismutase catalase and gpx compared with kidney or liver [ 1 ]. |
| 11179 | SOD1 | superoxide dismutase 1, soluble (amyotrophic lateral sclerosis 1 (adult)) | superoxide dismutase | 1.0 | to oxidative injury because of its high level of polyunsaturated fatty acids as substrates for lipid peroxidation high rate of oxygen consumption and low or moderate levels of the antioxidant enzymes superoxide dismutase catalase and gpx compared with kidney or liver [ 1 ]. |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | ho 1 | 1.0 | ho 1 mrna |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | ho 1 | 1.0 | the heme oxygenase 1 ho 1 mrna level was measured by northern blot analysis as previously described [ 21 ] using total cellular rna 5 _amp_#x3bc;g from control or ea treated nsc 34 cells about 2.5_amp_#xd7;10 6 cells . |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | heme oxygenase 1 | 1.0 | the heme oxygenase 1 ho 1 mrna level was measured by northern blot analysis as previously described [ 21 ] using total cellular rna 5 _amp_#x3bc;g from control or ea treated nsc 34 cells about 2.5_amp_#xd7;10 6 cells . |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | ho 1 | 1.0 | as a marker of oxidative stress we also measured the induction of ho 1 mrna. |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | ho 1 | 1.0 | exposure to 100 _amp_#x3bc;m ea promptly increased ho 1 mrna with a massive rise 3 h after treatment respectively 1.6 12 and 16 times the control level after 1 2 and 3 h of treatment; fig 5 . |
| 1516 | CAT | catalase | catalase | 1.0 | ease in dcfh da oxidation after treatment with ea indicates the presence of increased levels of h 2 o 2 and could be a direct consequence of gsh deficiency since in mitochondria_amp_#x2014;which lack catalase activity_amp_#x2014;h 2 o 2 is metabolized by gsh peroxidase using the reducing equivalents of gsh. |
| 1516 | CAT | catalase | catalase | 1.0 | in addition levels of catalase are very low in the nervous system [ 1 ]. |
| 1516 | CAT | catalase | catalase | 1.0 | interestingly treatment with catalase was beneficial in a mouse model of als [ 28 ]. |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | ho 1 | 1.0 | gsh depletion by ea induced expression of ho 1 which has been reported to be protective in different models of oxidative stress induced neuronal injury [ 43 and 44 ]. |
| 6876 | MAPK14 | mitogen-activated protein kinase 14 | p38 mitogen activated protein kinase | 1.0 | zymatic activity potentially act as antioxidants [ 45 ] and can thus exert anti apoptotic effects with co acting in a dominant manner as an anti apoptotic messenger possibly through activation of the p38 mitogen activated protein kinase mapk signal transduction pathway [ 46 ]. |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | ho 1 | 1.0 | the end products of ho 1 enzymatic activity potentially act as antioxidants [ 45 ] and can thus exert anti apoptotic effects with co acting in a dominant manner as an anti apoptotic messenger possibly through activation of t |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | ho 1 | 1.0 | however the role of ho 1 in our model requires further investigation since proapoptotic signals prevailed. |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | ho 1 | 1.0 | interestingly increased immunoreactivity for ho 1 and p38mapk were detected in human and mouse als [ 47 48 and 49 ]. |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | heme oxygenase 1 | 1.0 | effect of ethacrynic acid on heme oxygenase 1 expression in nsc 34 cells. |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | ho 1 | 1.0 | the heme oxygenase 1 ho 1 mrna signal is shown. |
| 5013 | HMOX1 | heme oxygenase (decycling) 1 | heme oxygenase 1 | 1.0 | the heme oxygenase 1 ho 1 mrna signal is shown. |