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Zinc and omega-3 mediates normalization of hepatic and cerebral cytochrome [P.sub.450s] in lindane-treated male rats.

Introduction

Lindane; [gamma]-hexachlorocyclohexane ([gamma]-HCH) an organochlorine insecticide and ubiquitous environmental contaminant in a potent neurostimulant including convulsions and other symptoms of hyperexcitability in mammals [1]. The residues of [gamma]-HCH has been shown to accumulate in the environment and gain entry human body through the food chain. Acute and chronic exposure of [gamma]-HCH has been shown to produce marked neurological and hepatotoxic effects in experimental animals [2, 3].

Lindane has been shown to be metabolized and accumulate in the brain. High concentrations of [gamma]-HCH in the brain have been related to its neurotoxic effects [4]. Metabolic studies have shown that HCH is biotransformed extensively by hepatic enzymes to form various metabolites [5]. In vitro studies have indicated that cytochrome [P.sub.450] enzymes are involved in the metabolism of [gamma]-HCH [5].

Role of zinc in modulating oxidative stress has recently been recognized. Oxidative stress is an important contributing factor in several chronic human diseases[6]. Together [O.sub.2.sup.-], [H.sub.2][O.sub.2] and[ .sup..]OH are known as reactive oxygen species (ROS) and these are produced continuously in vivo under aerobic conditions. The mitochondrial respiratory chain, microsomal cytochrome [P.sub.450] enzymes, flavoprotein oxidases, and peroxisomal fatty acid metabolism are the most significant intracellular sources of ROS [7].

The nicotinamide adenine dinucleotide phosphate (NADPH) oxidases are a group of plasma membrane associated enzymes, which catalyze the production of [O.sub.2.sup.-] from oxygen by using NADPH as the electron donor. Zinc is an inhibitor of this enzyme. The dismutation of [O.sub.2.sup.-] to [H.sub.2][O.sub.2] is catalyzed by an enzyme superoxide dismutase (SOD), which contains both copper and zinc. Zinc is known to induce the production of metallothionein, which is very rich in cysteine, and is an excellent scavenger of OH [8].

Omega-3 ([[omega].sub.3]) fatty acids are long chain, polyunsaturated fatty acids (PUFA) of plant and marine origin. Because these essential fatty acids (EFAs) cannot be synthesized in the human body, they must be derived from dietary sources. Flax seed, hemp, canola, and walnuts are generally rich sources of the [[omega].sub.3] PUFA alpha-linolenic acid (ALA). Fish provide varying amounts of [[omega].sub.3] fatty acids in the form of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA). ALA can be metabolized into the longer chain EPA and DHA. The role played by EFAs in the human body has been the subject of volumes of international research, particularly in recent years. The results indicate that [[omega].sub.3] fatty acids may be of value in the treatment of various medical conditions [9].

Thus, in this study we sought to investigate the protective potential of zinc and [[omega].sub.3] on drug metabolizing enzymes and lipid peroxidation in the rats liver and brain in lindane-induced toxicity.

Material and methods

Chemicals

Lindane; [gamma]-HCH (purity = 97%) was purchased from Aldrich Chemical. Reduced glutathione (GSH), 1-chloro-2,4-dinitrobenzene (CDNB), reduced nicotinamide adenine dinucleotide phosphate (NADPH), amidopyrine, cytochrome C, thiobarbituric acid and all other chemicals were purchased from Sigma Chemical Company (Saint Louis, USA). Use of lindane was approved by the Animal CareCompany Committee and met all guidelines for its use.

Animals

Forty male rats with average body weight of 200[+ or -]50 gm were obtained from National Research Institute, Cairo, Egypt and acclimatized for 2 weeks prior to the experiment. They were assigned to four groups and housed in Universal galvanized wire cages at room temperature (22-25 [degrees]C) and in a photoperiod of 14 h light/10 h dark per day. Animals received standard laboratory balanced commercial diet and water ad libitum.

Experimental design

The animals were housed in groups of 10 rats each and divided randomly into 4 groups. The first group served as control (C) and fed orally with corn oil, group 2 ([gamma]-HCH) rats treated with single dose of HCH (12 mg/kg, orally), 24 h prior to decapitation, group 3 ([[omega].sub.3]+Zn) fed orally with omega-3 (20 mg/kg) and zinc (0.84 mg/kg); treatment were carried out day by day for 20 days, group 4 ([gamma]-HCH+[[omega].sub.3]+Zn) treated with [gamma]-HCH (12 mg/kg, 24 h prior to decapitation), following the administration of [[omega].sub.3] and Zn day by day for 20 days.

Tissue preparations and assays Preparation of liver microsomes

Rats were fasted 24 h prior to each designated time point and then sacrificed by cervical dislocation. The abdominal cavity and the head were opened immediately, liver and brain were removed, washed with cold 0.1 mol phosphate buffer, pH 7.4, weighed and chilled on ice. All the following procedures were carried out in cold conditions. A 33% (W/V) crude homogenate was prepared in 0.1 mol phosphate buffer, pH 7.4 by homogenization with a Teflon pestle, using 5 strokes. The crud homogenate was then centrifuged at 11,000 xg for 20 min at 4 [degrees]C to remove the intact cells, nuclei and mitochondria. The supernatant solution was subsequently centrifuged at 105,000 xg for 60 min at 4 [degrees]C to sediment the microsomal pellet. The pellet was re-suspended in 0.1 mol phosphate buffer, pH 7.4, kept in ice bath and used as the enzyme source.

Protein determination

The protein concentration of the hepatic and brain microsomal fractions were determined by the method of Lowery et al. [10]

Enzyme assays

Liver and brain microsomal cytochrome [P.sub.450] and [b.sub.5] were determined according to Omura and Sato [11], using molar extinction coefficient 91 [cm.sup.-1] [mmol.sup.-1] for [P.sub.450] and 185 [cm.sup.-1] [mmol.sup.-1] for cytochrome [b.sub.5.] The activity of microsomal NADPH-cytochrome-C reductase was assayed according to the method of Williams and Kamin [12]. The total volume of incubation mixture was 2.2 ml and contained 0.5 mol potassium phosphate buffer, pH 7.5, 100 [eta]mol NADPH, 50 [eta]mol cytochrome C, and 33% microsomal enzyme. The rate of reduction of cytochrome C was measured at zero and 30 seconds after addition of NADPH at wavelength 550 [eta]m. The activity of this enzyme was calculated by using extinction coefficient of 21 [cm.sup.-1] m [mol.sup.-1.]

Glutathione-S-transferase activity was assayed according to the method of Habig Et-al. [13]. The incubation mixture contained 30 [micro]g protein of the supernatant fraction, 0.5 ml of reduced glutathione (0.5 mmol), 0.1 mol sodium phosphate buffer, pH 7.3. After preincubation at 37 [degrees]C for 5 min the reaction was initiated by adding 50 [micro]L of 1-chloro-2,4-dinitrobenzene (CDNB; 0.5 m mol) and incubated at 37 [degrees]C for another 5 min; the reaction was terminated by the addition of 0.2 ml of trichloroacetic acid solution (33% W/V). After centrifugation, the CDNB conjugate was measured spectrophotometrically at 340 [eta]m. Calculations were made using a molar extinction coefficient of 9.6 [cm.sup.-1] [mmol.sup.-1.] A unit of enzyme activity is defined as the amount of enzyme that catalyzes the formation of 1 [micro] mol of CDNB conjugate/mg protein/min under the assay conditions.

The activity of amidopyrine N-demethylase was measured according to Nash [14]. The method is based on measurement of the concentration of formaldehyde produced by oxidative N-demethylation of amidopyrine in microsome. The incubation mixture (1.71 ml) contained 0.4 ml of 0.1 mol Tris-HCl buffer pH 7.4, 0.4 ml of 2.5 [micro]mol magnesium chloride, 0.2 ml of 1 mmol NADPH, 0.1 ml of microsomal suspension and 0.11 ml of 80 mmol amidopyrine. After incubation at 37 [degrees]C for 20 min, the reaction was stopped by adding 0.25 ml of 25% zinc sulphate and 0.25 ml of aqueous solution of barium hydroxide. After centrifugation (1,000 xg for 10 min) formaldehyde was determined spectrophotometrically from changes in the color intensity of the supernatant at 412 [eta]m. The enzymatic activity was then expressed as [micro] mol of formaldehyde/min x kg liver/brain sample.

The activity of aniline 4-hydroxylase was measured according to Kato and Gillette[15]. The incubation mixture (1.71 ml) contained 0.4 ml of 0.08 mol Tris-HCl buffer, pH 7.4, 0.4 ml of 0.16 mol magnesium chloride, 0.2 ml of 0.03 mol NADPH, 0.1 ml of microsomal suspension and 0.11 ml of 0.03 mol aniline. After incubation at 37 [degrees]C for 20 min, the reaction was stopped by adding 0.5 ml of 15% trichloroacetic acid. After centrifugation (at 1,000 xg for 10 min), 1 ml of supernatant was added to 0.5 ml of 10% sodium carbonate and 1.5 ml of 2% phenol. After incubation at 37 [degrees]C for 30 min, the color developed was measured spectrophotometrically at 630 [eta]m.

Thiobarbituric acid reactive substances (TBARS) were measured according to the method described by Tapel and Zalkin [16]. The color intensity of the TBARS reactants was measured at 532 [eta]m and a molar extinction coefficient of 156,000 [cm.sup.-1] [mol.sup.-1] was used for calculation of the concentration.

Statistical analyses

Mean and standard error values were determined for all the parameters and the results were expressed as mean [+ or -] standard error for 10 rats in each group. The data were analyzed using a one-way analysis of variance (ANOVA). The student-Newmankeuls test was used to compare the treated and control groups and the significance is given as [rho]<0.05, [rho]<0.01 and [rho]<0.001.

Results

All the results of various treatment groups have been compared with their normal controls. However, results from [gamma]-HCH+[[omega].sub.3]+Zn treated group (group 4) have also been compared with the results of [gamma]-HCH treated group (group 2).

Clinical findings

Oral administration of a single dose of [gamma]-HCH (12 mg/kg) didn't cause mortality in the treated group. No symptoms of neurotoxicity were observed.

Microsomal protein and drug metabolizing enzymes

Hepatic and cerebral microsomal protein were significantly inhibited by 21 and 20%, respectively in [gamma]-HCH treated group compared to the control group. However, Zn and [[omega].sub.3] pretreatment to [gamma]-HCH -treated animals reversed the decreased microsomal protein (Table 1 and 2). Hepatic and cerebral microsomal [b.sub.5] and [P.sub.450] content were found to be significantly induced by 55 and 44% in liver, respectively and 35 and 17% in brain, respectively in [gamma]-HCH treated animals in comparison to untreated normal control. However, pretreatment of rats with Zn and [[omega].sub.3] prior to [gamma]-HCH administration reversed these increases in [b.sub.5] and [P.sub.450,] where complete normalization of cytochrome [b.sub.5] and [P.sub.450] were observed in [gamma]-HCH+[[omega].sub.3]+Zn treated group in liver, when compared to normal controls (Table 1 and 2).

The magnitude of induction in the activity of [P.sub.450] enzymes in rat brain was found to be much less as compared with liver. As shown in Table 1, [gamma]-HCH treatment resulted in a significant increase in hepatic microsomal NADPH cytochrome Creductase by 71%, while [gamma]-HCH decreased both amidopyrine N-demethylase and aniline-4-hydroxylase by 11 and 27%, respectively when compared to control. However, Zn and [[omega].sub.3] co-administration to [gamma]-HCH intoxicated animals normalized amidopyrine N-demethylase.

GST and lipid peroxidation

[gamma]-HCH treatment significantly increased hepatic and cerebral lipid peroxidation indicator; TBARS levels by 71 and 28%, respectively as compared to control value. Zn and [[omega].sub.3] supplementation to [gamma]-HCH treated animals brought back the already induced levels of TBARS (Table 1 and 2). GST was significantly enhanced in liver and brain of [gamma]-HCH treated animals as compared to control rats (Table 1 and 2).

Hepatic microsomal protein content was expressed as mg protein/ g liver, cytochrome [b.sub.5] and cytochrome [P.sub.450] contents were expressed as [eta]mol cytochrome/mg microsomal protein, glutathione S-transeferase activity was expressed as units/mg protein, NADPH cytochrome C-reductase activity was expressed as [eta]mol cytochrome C-reductase/mg protein/min, amidopyrine N-demethylase in liver microsomes was expressed as [micro]mol/min x kg liver sample, aniline-4-hydroxylase was expressed as [micro]mol/min/mg protein, and thiobarbituric acid reactive substances (TBARS) were expressed as [micro]mol TBARS/g tissue.Values are expressed as Mean [+ or -] SEM; n= 10 for each treatment group. [gamma]-HCH, lindane; [[omega].sub.3], omega-3; Zn, zinc. x, y, z, represent [rho]<0.05, 0.01, 0.001 compared with control respectively; a, b, c, represent [rho]< 0.05, 0.01, 0.001 in comparison between [gamma]-HCH and [gamma]-HCH +[[omega].sub.3]+Zn respectively.

Cerebral microsomal protein content was expressed as mg protein/ g brain, cytochrome [b.sub.5] and cytochrome [P.sub.450] contents were expressed as [eta]mol cytochrome/mg microsomal protein, glutathione S-transeferase activity was expressed as units/mg protein, NADPH cytochrome C-reductase activity was expressed as [eta]mol cytochrome C-reductase/mg protein/min, amidopyrine N-demethylase in brain microsomes was expressed as [micro]mol/min x kg brain sample, aniline-4-hydroxylase was expressed as [micro]mol/min/mg protein, and thiobarbituric acid reactive substances (TBARS) were expressed as [micro]mol TBARS/g tissue. [gamma]-HCH, lindane; [[omega].sub.3], omega-3; Zn, zinc.x, y, z, represent [rho]<0.05, 0.01, 0.001 compared with control respectively; a, b, c, represent [rho]< 0.05, 0.01, 0.001 in comparison between [gamma]-HCH and [gamma]-HCH +[[omega].sub.3]+Zn respectively.

Discussion

Hepatic microsomal [P.sub.450s] have been shown to play a significant role in the metabolism of [gamma]-HCH[17]. Lindane has been reported to induce several of the hepatic [P.sub.450]-dependent monooxygenases [18]. Liver microsomal induction of [P.sub.450s] by [gamma]-HCH can be considered as an adaptive response to increase the rate of [P.sub.450] metabolism for detoxification of the insecticide [19]. The induction observed in the activity of [P.sub.450] monooxygenases, specially in the brain, may be of toxicological significance due to the accumulation of significant amounts of [gamma]-HCH and its metabolites in the brain [20]. Even though the magnitude of induction of cerebral [P.sub.450s] is several fold less than the liver enzymes due to the presence of the blood-brain barrier [4]. Neurobehavioral studies have further demonstrated the role of [P.sub.450] induction and inhibition in the neurotoxicity of [gamma]-HCH. Even though self-induction in the hepatic [P.sub.450] monooxygenases appears to be important factor that minimizes the accumulation of [gamma]-HCH residues in the animal tissues [4].

In the present study, we observed an overall significant induction in the activity of [P.sub.450s] (except hepatic amidopyrine N-demethylase and aniline 4-hydroxylase activities). NADPH cytochrome C-reductase, a flavoprotein present in endoplasmic reticulum is thought to be involved in the oxidation of various drugs, steroids and other chemicals [21]. Alternatively, NADPH cytochrome C-reductase is also involved in initiating the process of NADPH-dependent lipid peroxidation in the microsomal membranes [22].

A significant inhibition of hepatic microsomal amidopyrine N-demethylase and aniline 4-hydroxylase activities observed in [gamma]-HCH intoxicated animals compared to control. Interestingly, we observed that co-administration of Zn and [[omega].sub.3] prior to [gamma]-HCH treatment to intoxicated animals reverted most of these altered enzymes levels to within normal limits. The simplistic explanation for these observed effects is that Zn regulates the hepatic microsomal drug metabolism, as well as the related oxidation of NADPH in the setting of [gamma]-HCH intoxication. Zn activates microsomal pyrophosphatase II, that is capable of metabolizing NADPH [23] Zinc is known to complex with NADPH, and it is suggested that regulation of drug metabolism by zinc might be due to this complex formation. It has also been hypothesized that Zn by binding to reductase, changes its oxidation-reduction potential, thus impacting the regulation of the cytochrome [P.sub.450] levels [24]. Additionally, it is known that majority of Zn in liver exists bound to zinc-metallothionein and this trace metal be readily available to the hepatic cytochromes [25].

Glutathione S-transferases (GSTs) form a group of enzymes that are present in high concentrations in the cytosol and catalyze a wide variety of substitution reactions in which glutathione (GSH) replaces an easily displaced group on the xenobiotic, and thus prevents the subsequent toxic reactions [26]. This reaction involves a compound with an electrophilic atom and GST facilitates the nucleophilic attack of glutathione thiolate on this electron deficient atom of the hydrophobic compound. GSH plays an important role in intracellular protection against toxic compounds, ROS, and free radicals. GSH protects the liver microsomes against the effects of reactive intermediates which are formed by cytochrome [P.sub.450] system as well as lipid peroxidation [27]. In the present study, the induction of GST activity in [gamma]-HCH intoxicated rats and the observed normalization of its activity following Zn and [[omega].sub.3] treatment this may refer to Zn property to induce metallothionein; is a free radical scavenger, or its indirect action in reducing the levels of ROS [27,28].

In conclusion, these data suggest that Zn and [[omega].sub.3] supplementation is protective in the animals subjected to [gamma]-HCH intoxication, as they markedly help regulating the activities of key drug metabolizing enzymes in conditions of lindane toxicity. Also, zinc-induced metallothionein levels and its antioxidant effects may be central to its biological protective role.

References

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[2] Barros, SB., Simizu, K., Junqueira, VB., 1991, Liver lipid peroxidationrelated parameters after short term administration of hexachlorohexane isomers to rats, Toxicol. Lett. , 56, pp. 137- 144.

[3] Smith, AG., 1991, Chlorinated hydrocarbons insecticides. In: Hayes Jr, WJ, Laws Jr, E.R. (Eds.), Handbook of Pesticide Toxicology Academic Press, San Diego; pp. 731-915.

[4] Parmar, D., Yadav, S., Dayal, M., Johri, A., Dhawan, A., Seth, PK., 2003, Effect of lindane on hepatic and brain cytochrome [P.sub.450s] and influence of [P.sub.450] modulation in lindane induced neurotoxicity, Food Chem .Toxicol., 41, pp. 1077- 1087.

[5] Baker, MT., Nelson, RM., Van Dyke, R., 1985, The information on chlorobenzene and benzene by the reductive metabolism of lindane in rat liver microsomes. Arch Biochem Biophys, 236, pp. 506- 514.

[6] Castro, L., Freeman, BA., 2001, Reactive oxygen species in human health and disease, Nutrition, 17, 161- 165.

[7] Lachance, PA., Nakat, Z., Jeong, W., 2001, Antioxidant: an integrative approach, Nutrition, 17, pp. 835- 838.

[8] Prasad, AS., 2008, Clinical, immunological, anti-inflammatory and antioxidant roles of zinc, Exp. Gerontol., 43, pp. 370- 377.

[9] Alan, C., Logan, ND., FRSH, MS., 2003, Neurobehavioral Aspects of Omega 3 Fatty Acids: Possible Mechanisms and Therapeutic Value in Major Depression, Altr .Med .Rev., 8, pp. 410-425.

[10] Lowery, OH., Rosebrough, NJ., Farr, AL., Randhll, RJ., 1951, Protein measurement with the Folin-phenol reagent, J. Biol .Chem., 193, pp.265- 275.

[11] Omura, T., Sato, R., 1964, The carbon monoxide binding pigment of liver microsomes. 1-Evidence for its hematoprotein nature, J. Biol .Chem., 239, pp.2370- 2378.

[12] Williams, CH., Kamin, H., 1962, Microsomal triphosphopyridine nucleotide-cytochrome-C reductase of liver, J. Biol. Chem. , 237, pp.587- 595.

[13] Habig, W., Pabst, M., Jakoby, W., 1974, Glutathione-S-transferases. The first enzymatic step in mercapturic acid formation, J. Biol. Chem., 249, pp. 7130-7139.

[14] Nash, T., 1953, The colorimetric estimation of formaldehyde by means of Hantzsch reactions, Biochem. J., 55, pp. 416- 421.

[15] Kato, R., Gillette, JR., 1965, Effects of starvation on NADPH-dependent enzymes in liver microsomes of male and female rats, J .Pharmacol. Exp. Therap., 150, pp. 279- 284.

[16] Tapel, AL., Zalkin, H., 1959, Inhibition of lipid peroxidation in mitochondria by vitamin E. Arch Biochem Biophys., 80, pp. 333- 336.

[17] Liu, PT., Morgan, DP., 1986, Coparative toxicity and biotransforamation of lindane in C57BL/6 and DBA/2 mice,Life Sci., 39, pp.1237- 1244.

[18] Puri, S., Kohli, KK., 1995, Differences in hepatic drug metabolizing enzymes and their response to lindane in rat, rabbit and monkey, Phamacol. Toxicol. , 77, pp.136- 141.

[19] Chadwick, RW., Copeland, MF., Mole, ML., Nesnow ,S., Cooke, N., 1981, Comparative effect of pretreatment with Phenobarbital, Arcolor 1254 and beta-naphthofalavone on the metabolism of lindane, Pest Biochem. Physiol., 15, pp. 120- 136.

[20] Sunol, C., Tussel, JM., Gelpi, E., Rodriguez-Farri, E., 1988, Convulsant effect of lindane and regional brain concentration of GABA and dopamine, Toxicology, 49, pp. 247-252.

[21] Masters, BS., 2005, The journey from NADPH cytochrome [P.sub.450] oxidoreductase to nitric oxide synthetases, Biochem. Biophys. Res. Commun., 338, pp.507- 519.

[22] Sevanian, A., Nordenbrand, K., Kim, E., Ernster, L., Hochstein, P., 1990, Microsomal lipid peroxidation: the role of NADPH-cytochrome [P.sub.450] reducatase and cytochrome [P.sub.450], Free Radic. Biol. Med., 8, pp. 145- 152.

[23] Zyryanov, AB., Tammenkoski, M., Salminen, A., Kolomiytseva, GY., Fabrichniy, IP., Goldman, A., Lahti, R., Baykov, AA., 2004, Site-specific effects of zinc on the activity of family II pyrophosphatase, Biochemistry, 43, pp. 14395-14402 .

[24] Zhou, Z., Wang, L., Song, Z., Saari, JT., McClain, CJ., Kang, YJ., 2005, Zinc supplementation prevents alcoholic liver injury in mice through attenuation of oxidative stress, Am. J. Pathol., 166, pp. 1681- 1690.

[25] Eaton, DL., Stacey, NH., Wong, KL., Klassen, CD., 1980, Dose-response effects of various metal ions on rat liver metallothionein, glutathione, heme oxygenase, and cytochrome [P.sub.450,] Toxicol. Appl. Pharmacol., 55, pp. 393-402.

[26] Siddiqui, MKJ., Mahboob, M., Mustafa, M., 1990, Hepatic and extrahepatic glutathione depletion and glutathione S-transferase inhibition by monocrotophos and thiol analogue, Toxicology, 64, pp. 271- 279.

[27] Sidhu, P., Garg, ML., Dhawan, DK., 2004, Protective effects of zinc on oxidative stress enzymes in liver of protein deficient rats, Nutr. Hosp., 19, pp.341- 347.

[28] Seagrave, J., Tobey, RA., Milderbrand, CE., 1983, Zinc effects on glutathione metabolism. Relationship to zinc induced protection from alkylating agents, Biochem. Pharmacol. , 32, pp. 3017- 3021.

Sabah G. El-Banna, Ahmed M. Attia and Mostafa M. Hussein

Department of Environmental Studies, Institute of Graduate Studies and Research, Alexandria University, Corresponding Author E-mail: sabah_gaber@yahoo.com
Table 1: Effects of omega3 and zinc on lindane induced oxidative
stress, on hepatic mixed function monooxygenases and TBARS of rat
liver.

Parameters Control [gamma]-HCH

Hepatic micro- 0.87 [+ or -] 0.08 0.69 [+ or -] 0.12(y)
somal protein
Cytochrome 2.47 [+ or -] 0.208 3.83 [+ or -] 0.26(z)
[b.sub.5]
Cytochrome 5.51 [+ or -] 0.289 7.91 [+ or -] 0.68(y)
[P.sub.450]
Glutathione 15.96 [+ or -] 0.990 20.73 [+ or -] 1.25(z)
S- transeferase
Cytochrome 852.1 [+ or -] 64.5 1455.0 [+ or -] 191.5(z)
C- reductase
Amidopyrine 0.186 [+ or -] 0.015 0.165 [+ or -] 0.002(x)
N-demethylase
Aniline- 0.165 [+ or -] 0.008 0.120 [+ or -] 0.011(y)
4-Hydroxylase
TBARS 2.29 [+ or -] 0.130 3.92 [+ or -] 0.244(z)

Parameters [[omega].sub.3]+Zn

Hepatic micro- 0.71 [+ or -] 0.14(y)
somal protein
Cytochrome 2.79 [+ or -] 0.26
[b.sub.5]
Cytochrome 4.78 [+ or -] 0.22(y)
[P.sub.450]
Glutathione 39.25 [+ or -] 1.32(z)
S- transeferase
Cytochrome 1115.7 [+ or -] 89.8(y)
C- reductase
Amidopyrine 0.177 [+ or -] 0.005
N-demethylase
Aniline- 0.320 [+ or -] 0.054(y)
4-Hydroxylase
TBARS 2.38 [+ or -] 0.077

Parameters [gamma]-HCH+
 [[omega].sub.3]+Zn

Hepatic micro- 1.06 [+ or -] 0.14(x,b)
somal protein
Cytochrome 2.46 [+ or -] 0.21(c)
[b.sub.5]
Cytochrome 5.34 [+ or -] 0.22(b)
[P.sub.450]
Glutathione 11.75 [+ or -] 1.06(z,c)
S- transeferase
Cytochrome 676.2 [+ or -] 27.6(y,c)
C- reductase
Amidopyrine 0.181 [+ or -] 0.005(b)
N-demethylase
Aniline- 0.081 [+ or -] 0.012(z,b)
4-Hydroxylase
TBARS 2.24 [+ or -] 0.109(c)

Table 2: Effects of omega3 and zinc on lindane induced oxidative
stress, on cerebral mixed function monooxygenases and TBARS of rat
brain.

Parameters Control [gamma]-HCH

Cerebral micro- 2.20 [+ or -] 0.24 1.75 [+ or -] 0.26(x)
somal protein
Cytochrome 2.62 [+ or -] 0.28 3.54 [+ or -] 0.27(z)
[b.sub.5]
Cytochrome 5.84 [+ or -] 0.10 6.86 [+ or -] 0.42(y)
[P.sub.450]
Glutathione 5.55 [+ or -] 0.495 10.54 [+ or -] 1.14(z)
S-transeferase
Cytochrome 316.5 [+ or -] 10.6 399.0 [+ or -] 14.5(z)
C-reductase
Amidopyrine 0.233 [+ or -] 0.006 0.222 [+ or -] 0.020
N-demethylase
Aniline- 0.030 [+ or -] 0.007 0.032 [+ or -] 0.005
4-Hydroxylase
TBARS 1.52 [+ or -] 0.028 1.94 [+ or -] 0.114(y)

Parameters [[omega].sub.3]+Zn [gamma]-HCH+
 [[omega].sub.3]+Zn

Cerebral micro- 2.40 [+ or -] 0.25 2.72 [+ or -] 0.37(x,c)
somal protein
Cytochrome 2.14 [+ or -] 0.15(x) 2.20 [+ or -] 0.23(y,c0
[b.sub.5]
Cytochrome 4.38 [+ or -] 0.43(z) 4.99 [+ or -] 0.37(y,b)
[P.sub.450]
Glutathione 11.10 [+ or -] 1.75(y) 5.17 [+ or -] 0.99(b)
S-transeferase
Cytochrome 292.3 [+ or -] 24.4(x) 264.4 [+ or -] 11.4(y,c)
C-reductase
Amidopyrine 0.227 [+ or -] 0.019 0.172 [+ or -] 0.006(z,b)
N-demethylase
Aniline- 0.040 [+ or -] 0.004(x) 0.026 [+ or -] 0.003(b)
4-Hydroxylase
TBARS 1.59 [+ or -] 0.140 1.82 [+ or -] 0.036(z)
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Author:El-Banna, Sabah G.; Attia, Ahmed M.; Hussein, Mostafa M.
Publication:International Journal of Biotechnology & Biochemistry
Date:Nov 1, 2010
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