Overexpression of manganese superoxide dismutase suppresses increases in collagen accumulation induced by culture of mesangial cells in high-media glucose
Superoxide has been implicated in the cellular signalling pathways, which regulate growth of mesangial cells (MC) and vascular smooth muscle cells. Manganese (Mn) 2+-dependent superoxide dismutase (SOD-2) is primarily responsible for metabolism of superoxide produced in mitochondria by respiratory c...
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Published in | Metabolism, clinical and experimental Vol. 50; no. 9; pp. 1043 - 1048 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
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New York, NY
Elsevier Inc
01.09.2001
Elsevier |
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Abstract | Superoxide has been implicated in the cellular signalling pathways, which regulate growth of mesangial cells (MC) and vascular smooth muscle cells. Manganese (Mn)
2+-dependent superoxide dismutase (SOD-2) is primarily responsible for metabolism of superoxide produced in mitochondria by respiratory chain activity during aerobic metabolism of glucose and other substrates. In the current studies, we examined the role of superoxide in the stimulation of collagen accumulation induced in MC by culture in media containing a high concentration of glucose. Aconitase, an iron sulfur enzyme whose activity is inhibited by superoxide, was used as an index of cellular superoxide production and action. SV-40[ndash ]transformed mouse MC were cultured in media containing 100 (low) or 500 (high) mg/dL
D-glucose and infected with a recombinant adenoviral (Ad) vector encoding either human mitochondrial Mn
2+ SOD-2 or green fluorescent protein (GFP). In cells infected with SOD-2 (SOD-2-Ad) and cultured in low glucose, SOD-2 activity was 5-fold higher than in cells infected with GFP (GFP-Ad), whereas Cu
2+/Zn
2+ cytoplasmic SOD (SOD-1) did not differ; culture in high-glucose media did not alter SOD-2 or SOD-1 activity in either GFD-Ad or SOD-2-Ad. In GFP-Ad, high glucose suppressed aconitase activity and increased collagen accumulation compared with corresponding values in low glucose. In SOD-2-Ad, high glucose failed to suppress aconitase activity or increase collagen accumulation. Addition of exogenous (presumably extracellular) SOD to GFP-Ad had no effect on the stimulation of collagen accumulation by high glucose. Analogous to the effects of SOD-2-Ad, diphenylene iodonium (DPI), a nonspecific inhibitor of the production of superoxide by mitochondrial respiration and other nicotinamide adenine dinucleotide (phosphate) (NAD)(P)H oxidase activities, reduced collagen accumulation in GFP-Ad cultured in low glucose and blocked stimulation of collagen accumulation induced by culture in high glucose. These results support a role for increased cellular superoxide production derived from NAD(P)H oxidase activity in the stimulation of collagen accumulation induced in MC by high glucose and demonstrate that an increase in mitochondrial SOD-2 activity suppresses this response. |
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AbstractList | Superoxide has been implicated in the cellular signalling pathways, which regulate growth of mesangial cells (MC) and vascular smooth muscle cells. Manganese (Mn)(2+)-dependent superoxide dismutase (SOD-2) is primarily responsible for metabolism of superoxide produced in mitochondria by respiratory chain activity during aerobic metabolism of glucose and other substrates. In the current studies, we examined the role of superoxide in the stimulation of collagen accumulation induced in MC by culture in media containing a high concentration of glucose. Aconitase, an iron sulfur enzyme whose activity is inhibited by superoxide, was used as an index of cellular superoxide production and action. SV-40-transformed mouse MC were cultured in media containing 100 (low) or 500 (high) mg/dL D-glucose and infected with a recombinant adenoviral (Ad) vector encoding either human mitochondrial Mn(2+) SOD-2 or green fluorescent protein (GFP). In cells infected with SOD-2 (SOD-2-Ad) and cultured in low glucose, SOD-2 activity was 5-fold higher than in cells infected with GFP (GFP-Ad), whereas Cu(2+)/Zn(2+) cytoplasmic SOD (SOD-1) did not differ; culture in high-glucose media did not alter SOD-2 or SOD-1 activity in either GFD-Ad or SOD-2-Ad. In GFP-Ad, high glucose suppressed aconitase activity and increased collagen accumulation compared with corresponding values in low glucose. In SOD-2-Ad, high glucose failed to suppress aconitase activity or increase collagen accumulation. Addition of exogenous (presumably extracellular) SOD to GFP-Ad had no effect on the stimulation of collagen accumulation by high glucose. Analogous to the effects of SOD-2-Ad, diphenylene iodonium (DPI), a nonspecific inhibitor of the production of superoxide by mitochondrial respiration and other nicotinamide adenine dinucleotide (phosphate) (NAD)(P)H oxidase activities, reduced collagen accumulation in GFP-Ad cultured in low glucose and blocked stimulation of collagen accumulation induced by culture in high glucose. These results support a role for increased cellular superoxide production derived from NAD(P)H oxidase activity in the stimulation of collagen accumulation induced in MC by high glucose and demonstrate that an increase in mitochondrial SOD-2 activity suppresses this response. Superoxide has been implicated in the cellular signalling pathways, which regulate growth of mesangial cells (MC) and vascular smooth muscle cells. Manganese (Mn) 2+-dependent superoxide dismutase (SOD-2) is primarily responsible for metabolism of superoxide produced in mitochondria by respiratory chain activity during aerobic metabolism of glucose and other substrates. In the current studies, we examined the role of superoxide in the stimulation of collagen accumulation induced in MC by culture in media containing a high concentration of glucose. Aconitase, an iron sulfur enzyme whose activity is inhibited by superoxide, was used as an index of cellular superoxide production and action. SV-40[ndash ]transformed mouse MC were cultured in media containing 100 (low) or 500 (high) mg/dL D-glucose and infected with a recombinant adenoviral (Ad) vector encoding either human mitochondrial Mn 2+ SOD-2 or green fluorescent protein (GFP). In cells infected with SOD-2 (SOD-2-Ad) and cultured in low glucose, SOD-2 activity was 5-fold higher than in cells infected with GFP (GFP-Ad), whereas Cu 2+/Zn 2+ cytoplasmic SOD (SOD-1) did not differ; culture in high-glucose media did not alter SOD-2 or SOD-1 activity in either GFD-Ad or SOD-2-Ad. In GFP-Ad, high glucose suppressed aconitase activity and increased collagen accumulation compared with corresponding values in low glucose. In SOD-2-Ad, high glucose failed to suppress aconitase activity or increase collagen accumulation. Addition of exogenous (presumably extracellular) SOD to GFP-Ad had no effect on the stimulation of collagen accumulation by high glucose. Analogous to the effects of SOD-2-Ad, diphenylene iodonium (DPI), a nonspecific inhibitor of the production of superoxide by mitochondrial respiration and other nicotinamide adenine dinucleotide (phosphate) (NAD)(P)H oxidase activities, reduced collagen accumulation in GFP-Ad cultured in low glucose and blocked stimulation of collagen accumulation induced by culture in high glucose. These results support a role for increased cellular superoxide production derived from NAD(P)H oxidase activity in the stimulation of collagen accumulation induced in MC by high glucose and demonstrate that an increase in mitochondrial SOD-2 activity suppresses this response. |
Author | Phillips, Stephen L. Melhem, Mona F. DeRubertis, Frederick R. Liachenko, Julia Craven, Patricia A. |
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Keywords | Culture medium Cell culture Enzyme Mesangial cell Superoxide dismutase Biosynthesis Glucose Gene expression Accumulation NADPH peroxidase Mitochondria Regulation(control) Peroxidases Blood vessel Collagen Circulatory system Oxidoreductases Manganese |
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Snippet | Superoxide has been implicated in the cellular signalling pathways, which regulate growth of mesangial cells (MC) and vascular smooth muscle cells. Manganese... |
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SubjectTerms | Aconitate Hydratase - metabolism Adenoviridae - genetics Animals Biological and medical sciences Blood vessels and receptors Cells, Cultured Collagen - metabolism Culture Media - pharmacology Enzyme Activation - drug effects Fundamental and applied biological sciences. Psychology Gene Expression Glomerular Mesangium - cytology Glomerular Mesangium - drug effects Glomerular Mesangium - metabolism Glucose - metabolism Glucose - pharmacology Green Fluorescent Proteins Humans Luminescent Proteins - biosynthesis Luminescent Proteins - genetics Mice NADPH Oxidases - metabolism Onium Compounds - pharmacology Superoxide Dismutase - biosynthesis Superoxide Dismutase - genetics Superoxide Dismutase-1 Transfection Transgenes Vertebrates: cardiovascular system |
Title | Overexpression of manganese superoxide dismutase suppresses increases in collagen accumulation induced by culture of mesangial cells in high-media glucose |
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