Effects of transfection with the Cu, Zn-superoxide dismutase gene on Xanthine/xanthine oxidase-induced cytotoxicity in fibroblasts from rat skin

Kohshi Nishiguchi, Kazue Ishida, Masanori Nakajima, Tetsuo Maeda, Fusao Komada, Seigo Iwakawa, Yusuke Tanigawara, Katsuhiko Okumura

研究成果: Article

6 引用 (Scopus)

抄録

Purpose. The effects of transfection with the human Cu, Zn-superoxide dismutase (hSOD)4 gene on active oxygen-induced cytotoxicity in rat skin fibroblasts (FR) were studied for the purpose of developing the novel delivery system of hSOD using hSOD gene. Methods. An expression plasmid for hSOD, pRc/RSV-SOD, was constructed and used to transfect FR cells. Xanthine (X)/xanthine oxidase (XO) system were used to generate active oxygen species. The effects of transfection with the hSOD gene on active oxygen-induced cytotoxicity were assessed by comparing the number of surviving cells and the level of lipid peroxidation in host and transformants after exposure to X/XO system. Results. The cellular SOD activity in RSV-SOD cells transfected with pRc/RSV-SOD was significantly increased in comparison with host or RSV cells transfected with the pRc/RSV plasmid containing no hSOD gene as a control. Furthermore, Western blot analysis using an anti-hSOD antibody indicated the production of hSOD in RSV-SOD cells. On the other hand, although the numbers of surviving cells in both host and RSV-SOD cultures after exposure to X/XO system decreased in a time-dependent manner, the decrease in number of surviving RSV-SOD cells was less than that in host cells. In the presence of catalase, the decreases in number of surviving cells in both host and RSV-SOD cultures after exposure to the X/XO system were also less than those in the absence of catalase. However, the decreases in cell survival in RSV-SOD cultures were significantly less than those in host cells in the presence of catalase. Furthermore, the levels of lipid peroxidation in RSV-SOD cells exposed to the X/XO system in the presence or absence of catalase were lower than those in host cells. These results indicated that the increase in cellular SOD activity by transfection with the hSOD gene protects cells from oxidative stress. Conclusions. Human SOD gene therapy may be useful for treatment of diseases in which oxidative tissue damage is produced.

元の言語English
ページ(範囲)577-582
ページ数6
ジャーナルPharmaceutical Research
13
発行部数4
DOI
出版物ステータスPublished - 1996
外部発表Yes

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Xanthine
Xanthine Oxidase
Fibroblasts
Cytotoxicity
Transfection
Rats
Skin
Genes
Catalase
Reactive Oxygen Species
Cell Count
Lipid Peroxidation
Plasmids
Lipids
Superoxide Dismutase
Superoxide Dismutase-1
Gene therapy
Oxidative stress
Cell culture
Genetic Therapy

ASJC Scopus subject areas

  • Chemistry(all)
  • Pharmaceutical Science
  • Pharmacology

これを引用

Effects of transfection with the Cu, Zn-superoxide dismutase gene on Xanthine/xanthine oxidase-induced cytotoxicity in fibroblasts from rat skin. / Nishiguchi, Kohshi; Ishida, Kazue; Nakajima, Masanori; Maeda, Tetsuo; Komada, Fusao; Iwakawa, Seigo; Tanigawara, Yusuke; Okumura, Katsuhiko.

:: Pharmaceutical Research, 巻 13, 番号 4, 1996, p. 577-582.

研究成果: Article

Nishiguchi, Kohshi ; Ishida, Kazue ; Nakajima, Masanori ; Maeda, Tetsuo ; Komada, Fusao ; Iwakawa, Seigo ; Tanigawara, Yusuke ; Okumura, Katsuhiko. / Effects of transfection with the Cu, Zn-superoxide dismutase gene on Xanthine/xanthine oxidase-induced cytotoxicity in fibroblasts from rat skin. :: Pharmaceutical Research. 1996 ; 巻 13, 番号 4. pp. 577-582.
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title = "Effects of transfection with the Cu, Zn-superoxide dismutase gene on Xanthine/xanthine oxidase-induced cytotoxicity in fibroblasts from rat skin",
abstract = "Purpose. The effects of transfection with the human Cu, Zn-superoxide dismutase (hSOD)4 gene on active oxygen-induced cytotoxicity in rat skin fibroblasts (FR) were studied for the purpose of developing the novel delivery system of hSOD using hSOD gene. Methods. An expression plasmid for hSOD, pRc/RSV-SOD, was constructed and used to transfect FR cells. Xanthine (X)/xanthine oxidase (XO) system were used to generate active oxygen species. The effects of transfection with the hSOD gene on active oxygen-induced cytotoxicity were assessed by comparing the number of surviving cells and the level of lipid peroxidation in host and transformants after exposure to X/XO system. Results. The cellular SOD activity in RSV-SOD cells transfected with pRc/RSV-SOD was significantly increased in comparison with host or RSV cells transfected with the pRc/RSV plasmid containing no hSOD gene as a control. Furthermore, Western blot analysis using an anti-hSOD antibody indicated the production of hSOD in RSV-SOD cells. On the other hand, although the numbers of surviving cells in both host and RSV-SOD cultures after exposure to X/XO system decreased in a time-dependent manner, the decrease in number of surviving RSV-SOD cells was less than that in host cells. In the presence of catalase, the decreases in number of surviving cells in both host and RSV-SOD cultures after exposure to the X/XO system were also less than those in the absence of catalase. However, the decreases in cell survival in RSV-SOD cultures were significantly less than those in host cells in the presence of catalase. Furthermore, the levels of lipid peroxidation in RSV-SOD cells exposed to the X/XO system in the presence or absence of catalase were lower than those in host cells. These results indicated that the increase in cellular SOD activity by transfection with the hSOD gene protects cells from oxidative stress. Conclusions. Human SOD gene therapy may be useful for treatment of diseases in which oxidative tissue damage is produced.",
keywords = "Catalase, Gene therapy, Human Cu, Oxidative stress, Zn-SOD",
author = "Kohshi Nishiguchi and Kazue Ishida and Masanori Nakajima and Tetsuo Maeda and Fusao Komada and Seigo Iwakawa and Yusuke Tanigawara and Katsuhiko Okumura",
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TY - JOUR

T1 - Effects of transfection with the Cu, Zn-superoxide dismutase gene on Xanthine/xanthine oxidase-induced cytotoxicity in fibroblasts from rat skin

AU - Nishiguchi, Kohshi

AU - Ishida, Kazue

AU - Nakajima, Masanori

AU - Maeda, Tetsuo

AU - Komada, Fusao

AU - Iwakawa, Seigo

AU - Tanigawara, Yusuke

AU - Okumura, Katsuhiko

PY - 1996

Y1 - 1996

N2 - Purpose. The effects of transfection with the human Cu, Zn-superoxide dismutase (hSOD)4 gene on active oxygen-induced cytotoxicity in rat skin fibroblasts (FR) were studied for the purpose of developing the novel delivery system of hSOD using hSOD gene. Methods. An expression plasmid for hSOD, pRc/RSV-SOD, was constructed and used to transfect FR cells. Xanthine (X)/xanthine oxidase (XO) system were used to generate active oxygen species. The effects of transfection with the hSOD gene on active oxygen-induced cytotoxicity were assessed by comparing the number of surviving cells and the level of lipid peroxidation in host and transformants after exposure to X/XO system. Results. The cellular SOD activity in RSV-SOD cells transfected with pRc/RSV-SOD was significantly increased in comparison with host or RSV cells transfected with the pRc/RSV plasmid containing no hSOD gene as a control. Furthermore, Western blot analysis using an anti-hSOD antibody indicated the production of hSOD in RSV-SOD cells. On the other hand, although the numbers of surviving cells in both host and RSV-SOD cultures after exposure to X/XO system decreased in a time-dependent manner, the decrease in number of surviving RSV-SOD cells was less than that in host cells. In the presence of catalase, the decreases in number of surviving cells in both host and RSV-SOD cultures after exposure to the X/XO system were also less than those in the absence of catalase. However, the decreases in cell survival in RSV-SOD cultures were significantly less than those in host cells in the presence of catalase. Furthermore, the levels of lipid peroxidation in RSV-SOD cells exposed to the X/XO system in the presence or absence of catalase were lower than those in host cells. These results indicated that the increase in cellular SOD activity by transfection with the hSOD gene protects cells from oxidative stress. Conclusions. Human SOD gene therapy may be useful for treatment of diseases in which oxidative tissue damage is produced.

AB - Purpose. The effects of transfection with the human Cu, Zn-superoxide dismutase (hSOD)4 gene on active oxygen-induced cytotoxicity in rat skin fibroblasts (FR) were studied for the purpose of developing the novel delivery system of hSOD using hSOD gene. Methods. An expression plasmid for hSOD, pRc/RSV-SOD, was constructed and used to transfect FR cells. Xanthine (X)/xanthine oxidase (XO) system were used to generate active oxygen species. The effects of transfection with the hSOD gene on active oxygen-induced cytotoxicity were assessed by comparing the number of surviving cells and the level of lipid peroxidation in host and transformants after exposure to X/XO system. Results. The cellular SOD activity in RSV-SOD cells transfected with pRc/RSV-SOD was significantly increased in comparison with host or RSV cells transfected with the pRc/RSV plasmid containing no hSOD gene as a control. Furthermore, Western blot analysis using an anti-hSOD antibody indicated the production of hSOD in RSV-SOD cells. On the other hand, although the numbers of surviving cells in both host and RSV-SOD cultures after exposure to X/XO system decreased in a time-dependent manner, the decrease in number of surviving RSV-SOD cells was less than that in host cells. In the presence of catalase, the decreases in number of surviving cells in both host and RSV-SOD cultures after exposure to the X/XO system were also less than those in the absence of catalase. However, the decreases in cell survival in RSV-SOD cultures were significantly less than those in host cells in the presence of catalase. Furthermore, the levels of lipid peroxidation in RSV-SOD cells exposed to the X/XO system in the presence or absence of catalase were lower than those in host cells. These results indicated that the increase in cellular SOD activity by transfection with the hSOD gene protects cells from oxidative stress. Conclusions. Human SOD gene therapy may be useful for treatment of diseases in which oxidative tissue damage is produced.

KW - Catalase

KW - Gene therapy

KW - Human Cu

KW - Oxidative stress

KW - Zn-SOD

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