Abstract
To establish a simple screening system for estimating efficacy of an agent for an oxidative-related lesion, we investigated the damage in isolated rat hepatocytes exposed to 75 μM tert-butyl hydroperoxide (t-BuOOH) and then subsequently incubated the cells in fresh medium. By electron.spin resonance spectroscopy analysis using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), DMPO adducts of tert-butoxyl radicals and carbon center radicals were detected during the t-BuOOH exposure, and DMPO-OH formation was detected after t-BuOOH removal. In t-BuOOH-exposed cells, the level of phosphatidylcholine hydroperoxide (PCOOH), a peroxidative product of biomembranes in the hepatocytes, and the leakage of enzymes into the culture medium were significantly increased. An increase in acid phosphatase (AP) activity representing lysosome destabilization preceded the aspartate oxoglutarate aminotransferase (AST), alanine oxoglutarate aminotransferase (ALT) and lactate dehydrogenase (LDH) leakage. Ninjin-yoei-to added to the culture medium following the t-BuOOH exposure significantly inhibited the PCOOH formation and the leakage of AP, AST, ALT and LDH, concentration-dependently. Ninjin-yoei-to at 1 mg/ml in culture medium completely diminished these increases in enzyme activities down to the background levels found in control experiments and this reduction was greater than the most effective α-tocopherol concentration of 20 μmol/ml. Considering all of these results, it is likely Ninjin-yoei-to may exert its protective effect by anti-oxidative action and membrane stabilization.
| Original language | English |
|---|---|
| Pages (from-to) | 227-233 |
| Number of pages | 7 |
| Journal | Japanese Journal of Pharmacology |
| Volume | 85 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - 2001 |
| Externally published | Yes |
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Keywords
- Biomembrane peroxidation
- Isolated hepatocyte
- Lysosome
- Ninjin-yoei-to
- Oxidative stress
ASJC Scopus subject areas
- Pharmacology
- Molecular Medicine
Cite this
Protective effect of Ninjin-yoei-to on damage to isolated hepatocytes following transient exposure to tert-butyl hydroperoxide. / Takayama, Fusako; Egashira, T.; Yamanaka, Y.
In: Japanese Journal of Pharmacology, Vol. 85, No. 3, 2001, p. 227-233.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Protective effect of Ninjin-yoei-to on damage to isolated hepatocytes following transient exposure to tert-butyl hydroperoxide
AU - Takayama, Fusako
AU - Egashira, T.
AU - Yamanaka, Y.
PY - 2001
Y1 - 2001
N2 - To establish a simple screening system for estimating efficacy of an agent for an oxidative-related lesion, we investigated the damage in isolated rat hepatocytes exposed to 75 μM tert-butyl hydroperoxide (t-BuOOH) and then subsequently incubated the cells in fresh medium. By electron.spin resonance spectroscopy analysis using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), DMPO adducts of tert-butoxyl radicals and carbon center radicals were detected during the t-BuOOH exposure, and DMPO-OH formation was detected after t-BuOOH removal. In t-BuOOH-exposed cells, the level of phosphatidylcholine hydroperoxide (PCOOH), a peroxidative product of biomembranes in the hepatocytes, and the leakage of enzymes into the culture medium were significantly increased. An increase in acid phosphatase (AP) activity representing lysosome destabilization preceded the aspartate oxoglutarate aminotransferase (AST), alanine oxoglutarate aminotransferase (ALT) and lactate dehydrogenase (LDH) leakage. Ninjin-yoei-to added to the culture medium following the t-BuOOH exposure significantly inhibited the PCOOH formation and the leakage of AP, AST, ALT and LDH, concentration-dependently. Ninjin-yoei-to at 1 mg/ml in culture medium completely diminished these increases in enzyme activities down to the background levels found in control experiments and this reduction was greater than the most effective α-tocopherol concentration of 20 μmol/ml. Considering all of these results, it is likely Ninjin-yoei-to may exert its protective effect by anti-oxidative action and membrane stabilization.
AB - To establish a simple screening system for estimating efficacy of an agent for an oxidative-related lesion, we investigated the damage in isolated rat hepatocytes exposed to 75 μM tert-butyl hydroperoxide (t-BuOOH) and then subsequently incubated the cells in fresh medium. By electron.spin resonance spectroscopy analysis using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), DMPO adducts of tert-butoxyl radicals and carbon center radicals were detected during the t-BuOOH exposure, and DMPO-OH formation was detected after t-BuOOH removal. In t-BuOOH-exposed cells, the level of phosphatidylcholine hydroperoxide (PCOOH), a peroxidative product of biomembranes in the hepatocytes, and the leakage of enzymes into the culture medium were significantly increased. An increase in acid phosphatase (AP) activity representing lysosome destabilization preceded the aspartate oxoglutarate aminotransferase (AST), alanine oxoglutarate aminotransferase (ALT) and lactate dehydrogenase (LDH) leakage. Ninjin-yoei-to added to the culture medium following the t-BuOOH exposure significantly inhibited the PCOOH formation and the leakage of AP, AST, ALT and LDH, concentration-dependently. Ninjin-yoei-to at 1 mg/ml in culture medium completely diminished these increases in enzyme activities down to the background levels found in control experiments and this reduction was greater than the most effective α-tocopherol concentration of 20 μmol/ml. Considering all of these results, it is likely Ninjin-yoei-to may exert its protective effect by anti-oxidative action and membrane stabilization.
KW - Biomembrane peroxidation
KW - Isolated hepatocyte
KW - Lysosome
KW - Ninjin-yoei-to
KW - Oxidative stress
UR - http://www.scopus.com/inward/record.url?scp=0035067888&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0035067888&partnerID=8YFLogxK
U2 - 10.1254/jjp.85.227
DO - 10.1254/jjp.85.227
M3 - Article
C2 - 11325014
AN - SCOPUS:0035067888
VL - 85
SP - 227
EP - 233
JO - Japanese Journal of Pharmacology
JF - Japanese Journal of Pharmacology
SN - 0021-5198
IS - 3
ER -