Phosphatidylcholine Hydroperoxide (PCOOH) And Phosphatidylethanolamine Hydroperoxide Research Articles

Lipid peroxidation during ischemia depends on ischemia time in warm ischemia and reperfusion of rat liver

Prolonged hepatic warm ischemia has been incriminated in oxidative stress after reperfusion. However, the magnitude of oxidative stress during ischemia has been controversial. The aims of the present study were to elucidate whether lipid peroxidation progressed during ischemia and to clarify whether oxidative stress during ischemia aggravated the oxidative damage after reperfusion. Rats were subjected to 30 to 120 min of 70% warm ischemia alone or followed by reperfusion for 60 min. Lipid peroxidation (LPO) was evaluated by amounts of phosphatidylcholine hydroperoxide (PC-OOH) and phosphatidylethanolamine hydroperoxide (PE-OOH) as primary LPO products. Total amounts of malondialdehyde and 4-hydroxy-2-nonenal (MDA + 4-HNE), degraded from hydroperoxides, were also determined. PC-OOH and PE-OOH significantly increased at 60 and 120 min ischemia with concomitant increase of oxidized glutathione. These hydroperoxides did not increase at 60 min reperfusion after 60 min ischemia, whereas they did increase at 60 min reperfusion after 120 min ischemia with deactivation of phospholipid hydroperoxide glutathione peroxidase and superoxide dismutase. The amount of MDA + 4-HNE exhibited similar changes, but the velocity of production dropped with ischemic time longer than 60 min. In conclusion, oxidative stress progressed during ischemia and triggered the oxidative injury after reperfusion. Secondary LPO products are less sensitive, especially during ischemia, which may cause possible underestimation and discrepancy.

Suppression of lipid-hydroperoxide and DNA-adduct formation by isoflavone-containing soy hypocotyl tea in rats

Phytoestrogen isoflavones (IFs) are considered to suppress estrogen-related cancers through their antiestrogenic activity. The antioxidant effect of IFs, however, has not been confirmed in anin vivo system, so suppression of hydroperoxide formation and resultant DNA adduct formation were studied. The antioxidant effects of the soya-hypocotyl tea (SHT), which contained daidzein (14+/-1.5 mg/l) and genistein (3+/-0.5 mg/l), were examined in Wistar rats fed the AIN-76 control diet or iron deficient diet (FeD) for 4 weeks. The intake amount of the diet and IFs were measured daily. Urinary excretion of IFs was measured for 3 days before sacrifice. In addition to the serum lipid analyses, phosphatidylcholine hydroperoxide (PCOOH), and phosphatidylethanolamine hydroperoxide (PEOOH) production in red blood cells and the liver were measured as a biomarker of oxidants. Production of DNA adducts by oxidative stress was measured by the amount of 8-hydroxy-2'-deoxyguanosine (oh(8)dG) in the liver and kidney, and urine. Histological changes were checked by H&E staining and immunohistochemistry for oh(8)dG. FeD rats showed anemia, growth retardation, hyperlipidemia. IFs only lowered the triacylglycerol level and did not change the cholesterol level. Rats fed the normal diet did not show suppression of PCOOH and PEOOH production in either red blood cells or the liver, while groups administered SHT showed suppressed production of PCOOH and PEOOH in the liver. The cumulative intake of daidzein, genistein and the total amount of IFs showed significant inverse associations with urinary excretion of oh(8)dG. oh(8)dG in the kidney showed an inverse association with the amount of oh(8)dG in the urine. Enzymehistochemically, a strong localization of oh(8)dG was found in the epithelial cells of the bile canaliculi and proximal tubules of the kidney. IFs and SHT showed antioxidant effects at physiological concentrations in anin vivo system. The antioxidant effects of IFs decreased oxidation stress to the nuclear DNA, which was shown by the decreased oh(8)dG production. It is suggested that to prevent various cancers, in addition to the known antiestrogenie, antityrosin kinase, and other effects. IFs appeared to promote excretion of oh(8)dG.

Antioxidant activity of soya hypocotyl tea in humans.

Antioxidative activity of isoflavones has not been shown in humans. Newly-developed isoflavone-rich soya hypocotyl tea contains about 12 mg isoflavones per liter. 15 tea drinkers and 23 control young female students were randomly selected from volunteers, and underwent physical examination, blood chemistry and urinary analysis before and after one month of tea drinking. A three-day dietary record was taken before each physical examination. The tea drinkers showed a lower level of phosphatidylcholine hydroperoxide (PCOOH) and phosphatidyl-ethanolamine hydroperoxide (PEOOH) in the red blood cells and a significant reduction of 8-hydroxydeoxyguanine (8ohdG) in the urine compared to the controls.

Abstracts of literature

Abstracts of literature

Effects of unilateral intrathecal administrations of low dose tissue-type plasminogen activator on clot lysis, vasospasm and brain phospholipid hydroperoxidation in a primate model of bilateral subarachnoid hemorrhage

In order to clarify the effect of clot lysis by recombinant tissue-type plasminogen activator (tPA) on the brain lipid peroxidationl we measured phosphatidylcholine hydroperoxide (PCOOH) and phosphatidylethanolamine hydroperoxide (PEOOH) levels in a primate model of subarachnoid hemorrhage (SA H). Monkeys were assigned into two groups; a tPA-treated group receiving intrathecal injections of 0.02 mg tPAI and a placebo-treated group receiving saline. The tPA or placebo was injected into the right side of the basal cistern every 8 h for 6 days following bilateral SAH induction. The tPA cleared the right side clots (p < 0.0007)1 but not the left side clots. The degree of vasospasm in the right middle cerebral artery and the rCBF decrease in the right parietal cortex were significantly attenuated in the tPA group (p < 0.05). In the placebo groupi more severe vasospasm and marked rCBF reduction were noted in comparison with those in the tPA group. PCOOH levels in the parietal cortex were significantly higher in the placebo group than in the tPA group (p < 0.05). There were no significant changes in brain PEOOH levels. These results may explain the limitations for clinical application of unilateral intrathecal administration of tPA. [Neural Res 1998; 20: 625–631]

Age-associated oxidative damage in microsomal and plasma membrane lipids of rat hepatocytes

Phosphatidylcholine hydroperoxide (PC-OOH) and phosphatidylethanolamine hydroperoxide (PE-OOH) concentrations were determined in microsomes and plasma membranes prepared from 2- and 17-month-old male Sprague-Dawley rat hepatocytes, to verify the dissimilarity of age dependency of lipid peroxidation in organelle membranes. The hydroperoxides were directly measured by chemiluminescence detection–high-performance liquid chromatography (CL–HPLC), and 1-palmitoyl-2-(13-hydroperoxy- cis-9, trans-11-octadecadienoyl) phosphatidylcholine (PLPC-OOH) and 1-palmitoyl-2-(13-hydroperoxy- cis-9, trans-11-octadecadienoyl) phosphatidylethanolamine (PLPE-OOH) were enzymatically synthesized and utilized as standards for the calibration. Baseline concentrations of hydroperoxides (PC-OOH+PE-OOH) of the 17-month-old rats were 46 pmol per mg protein in microsomes (2.7 times higher than the 2-month-old rats) and 306 pmol per mg protein in plasma membranes (9.9 times higher than the 2-month-old rats). Both microsomal and plasma membrane lipids were severely peroxidized and converted to phospholipid hydroperoxides by NADPH-dependent lipid peroxidation in vitro, but the age-dependency was only observed in the plasma membranes. These results demonstrate that substantial oxidative damage to membrane phospholipids occurs with ageing both in microsomes and plasma membranes, but is more prevalent in plasma membranes in rat hepatocytes.

Cytotoxicity of organochlorine pesticides and lipid peroxidation in isolated rat hepatocytes.

The cytotoxicity and lipid peroxidation of pesticides containing a halogen group were examined in isolated rat hepatocytes. We examined 9 pesticides of 3 different representative chemical families, chlorinated aromatic fungicides (pentachlorophenol (PCP), pentachloronitrobenzene (PCNB), chlorothalonil, fthalide), polyhaloalkylated thio fungicides (dichlofluanid, captan) and diphenyl ether herbicide (2,4-dichlorophenyl 4-nitrophenyl ether (NIP), 4-nitrophenyl2,4,6-trichlorophenyl ether (CNP), chlomethoxynil). The contents of the hydroperoxides in phospholipid, phosphatidylcholine hydroperoxide (PCOOH) and phosphatidylethanolamine hydroperoxide (PEOOH) were determined by the HPLC-chemiluminescence (CL-HPLC) method, which is sensitive and specific for lipid hydroperoxide. Chlorothalonil, dichlofluanid and captan were the most potent cytotoxicants evaluated by lactate dehydrogenase (LDH) leakage. PCP, NIP and CNP exhibited intermediate cytotoxicity. PCNB, fthalide and chlomethoxynil showed low cytotoxicity. The cellular phospholipid hydroperoxide (PCOOH and PEOOH) levels were remarkably increased by chlorothalonil (PCOOH, 23 times and PEOOH, 7 times), dichlofluanid (PCOOH, 523 times and PEOOH, 22 times) and captan (PCOOH, 518 times and PEOOH, 16 times) as compared with the control group. The PCOOH content was slightly increased by PCP (4.8 times) and NIP (6.3 times), whereas the other 4 pesticides did not change the phospholipid hydroperoxide level. Severe cytotoxicity was observed with a remarkable increase of phospholipid hydroperoxide by chlorothalonil, dichlofluanid and captan.

Age-related change of phosphatidylcholine hydroperoxide and phosphatidylethanolamine hydroperoxide levels in normal human red blood cells

The age-related occurrence of phosphatidylcholine hydroperoxide (PCOOH) and phosphatidylethanolamine hydroperoxide (PEOOH) in normal human red blood cells (RBC) was confirmed by using chemiluminescence detection-high performance liquid chromatography (CL-HPLC). The concentration (mean ± S.D.) for the healthy young adult (22–27 of age. n = 20) was 93 ± 17 pmol PCOOH and 121 ± 20 pmol PEOOH/ml packed RBC, while for the aged adult (56–92 of age, n = 20) the peroxide content was significantly higher, 162 ± 52 pmol PCOOH and 186 ± 40 pmol PEOOH/ml packed RBC. These results indicate that oxidative stress occurs constantly on RBC, even in normal humans, and that the susceptivity significantly increases with age.

Cytotoxicity of Trihalomethanes and Lipid Peroxidation in Isolated Rat Hepatocytes.

Cytotoxic and lipid peroxidative effects of 4 major trihalomethanes (chloroform, CHCl3 ; bromodichloromethane, BrCHCl2 ; chlorodibromomethane, ClCHBr2 ; bromoform, CHBr3) were examined in isolated rat hepatocytes. The phospholipid hydroperoxides, phosphatidylcholine hydroperoxide (PCOOH) and phosphatidylethanolamine hydroperoxide (PEOOH), were determined by the HPLC-chemiluminescence detection (CL-HPLC) method which is sensitive and specific for lipid hydroperoxide. Severe cytotoxicity was observed when the cells were cultured with 50 mM of CHCl3, 20 mM of BrCHCl2, 10 mM of ClCHBr2 and 10 mM of CHBr3, respectively. At these concentrations, cytotoxicity was observed after 10 min or more incubation with each 4 trihalomethane. The cellular lipid hydroperoxide was remarkably increased with the cytotoxicity by BrCHCl2 and the other three trihalomethanes induced slight lipid peroxidation. The cytotoxicity was not effectively prevented by α-tocopherol, though the lipid peroxidation was appreciably prevented. These results suggested that trihalomethane induces lipid peroxidation in isolated rat hepatocytes, but that this may not be the main cause of the cytotoxicity.

Cytotoxicity of chlorinated hydrocarbons and lipid peroxidation in isolated rat hepatocytes.

The effects of 11 chlorinated hydrocarbons on toxicological parameters and lipid peroxidation were examined in cultured rat hepatocytes. The cytotoxicity was evaluated by measuring the lactate dehydrogenase (LDH) released and the protein amount in cells attached to culture dishes. Phosphatidylcholine hydroperoxide (PCOOH) and phosphatidylethanolamine hydroperoxide (PEOOH) levels were determined by HPLC with chemiluminescence detection (CL-HPLC). Severe cytotoxicity was observed under the presence of the carbon tetrachloride, 1,1,1-trichloroethane, tetrachloroethylene and 1,3-dichloropropene in a concentration of 10 mM. The cellular PCOOH and PEOOH levels were remarkably increased (more than 4 times) by adding these 4 chlorinated hydrocarbons. The other 7 chlorinated hydrocarbons examined did not cause any change on the cellular phospholipid hydroperoxide levels, and showed lower or no cytotoxicity at a concentration of 10 mM. The peroxidative degradation of membrane phospholipids may play an important role in causing the cytotoxicity of some chlorinated hydrocarbons.

Chemiluminescent simultaneous determination of phosphatidylcholine hydroperoxide and phosphatidylethanolamine hydroperoxide in the liver and brain of the rat.

The quantification of phospholipid hydroperoxides in biological tissues is important in order to know the degree of peroxidative damage of membrane lipids. For this purpose, optimal conditions for the chemiluminescent simultaneous assay of phosphatidylcholine hydroperoxide (PCOOH) and phosphatidylethanolamine hydroperoxide (PEOOH) in rat liver and brain were determined. A chemiluminescence detection-high performance liquid chromatography (CL-HPLC) method that incorporates cytochrome c and luminol as a post-column hydroperoxide-specific luminescent reagent was used (Miyazawa et al. 1987. Anal. Lett. 20: 915-925; Miyazawa. 1989. Free Radical Biol. Med. 7: 209-217). An n-propylamine-bound silica column with hexane-2-propanol-methanol-water 5:7:2:1 (v/v/v/v) (flow rate 1.0 ml/min) as eluant was used to determine both PCOOH and PEOOH, which were separated from each other and from other lipids and lipid-soluble antioxidants. High reproducibility and sensitivity as low as 10 pmol hydroperoxide-O2 were observed with a mixture of 10 micrograms/ml cytochrome c and 2 micrograms/ml luminol in 50 mM borate buffer (pH 10.0, flow rate 1.1 ml/min) as luminescent reagent and a post-column mixing joint temperature of 40 degrees C. Using the established analytical conditions, it was confirmed that both PCOOH (1324 +/- 122 pmol/g liver, 114 +/- 18 pmol/g brain, mean +/- SD) and PEOOH (728 +/- 89 pmol/g liver, 349 +/- 60 pmol/g brain, mean +/- SD) are present in the liver and brain of Sprague-Dawley rats bred on a slightly modified AIN-76A semisynthetic diet for 3 months. The phospholipid hydroperoxide content in the rat liver was shown to be affected by dietary oils, but not significantly affected in the brain.