Www.ejmanager.com



Potential Protective Effect of Ginseng on Dexamethasone-Induced Liver Damage Via Motivation of Anti-Apoptotic and Antioxidant Activities in RatsWafaa E. Elkhrashy, Mohamed A. Lebda, Abd El-Wahab A. Mandour , Nabil M. Taha and Aml S. Hashem*Biochemistry Department, Faculty of Veterinary Medicine, Alexandria University* Corresponding author: Aml S. Hashem, amlhashem2009@Running title: Ginseng and dexamethasone liver damageABSTRACTThe current study aimed to explore the protective roles of ginseng against dexamethasone liver devastating effects. Sixty male rats were equally allocated into six groups; G1: control, G2: ginseng aqueous extract (200 mg/kg B.W. orally /4 weeks), G3: dexamethasone (1mg/kg B.W. i.p/one week then decapitated), G4: dexamethasone- withdrawal group (1mg/kg B.W. i.p/one week and decapitated at the end of experiment), G5: dexamethasone ginseng treated group (injected with dexamethasone/one week then treated with ginseng/other 3weeks), and G6: ginseng-dexamethasone-ginseng group (received ginseng/two weeks followed by dexamethasone/one week then ginseng/another week). Dexamethasone significantly increased serum liver enzymatic activities alongside with mild to moderate hepatic vacuolation, and periportal mononuclear infiltration suggesting liver damage. The level of hepatic malondialdehyde (MDA) was elevated markedly with subsequent decline in the reduced glutathione concentration following challenged with dexamethasone. Moreover, the hepatic expression of thioredoxin mRNA transcript was down-regulated while BCL2-associated X protein (Bax) mRNA transcript was up-regulated providing imbalance in antioxidant and apoptosis pathway. Ginseng treatment ameliorated the damaged effects of dexamethasone on the liver as indicated by attenuation of elevated liver enzymatic activities, augmentation the reduced glutathione level, up-regulation thioredoxin mRNA transcription along with down-regulation of Bax mRNA expression in liver tissue. Conclusively, ginseng exerted a protective action on dexamethasone hepatic side effects through anti-apoptotic and antioxidant properties.Key words:Antioxidant; Apoptosis; Dexamethasone; Glutathione; Hepatic damage; Thioredoxin1- INTRODUCTION Glucocorticoids are one of the most prescribed drugs for a huge number of inflammatory and immunological diseases (Duma et al., 2006). In spite of its therapeutic activity, high doses with long-term consumption of these medications are associated with serious side effects, such as diabetes mellitus, liver disorders, cardiovascular disorders, hypertension, dyslipidemia, and osteoarthritis (Duma et al., 2006). Dexamethasone is a catabolic rather than anabolic steroid enhancing the catabolism of stored carbohydrates, lipids, and proteins during stress conditions (Lingaiah et al., 2012). Dexamethasone can cross the cell membrane binding to its cytoplasmic receptors forming hormone receptor complex that enters the cell nucleus stimulating the transcription of specific mRNA and subsequently synthesis of enzymes responsible for systematic effects of corticosteroids (Dolatabadi and Zarchii, 2015).Panax ginseng, herbal medicine, used to fight stress in China, Japan and Korea for more than 200 years. Ginseng is composed of various components including ginsenosides, panaxosides, volatile oil, sterols, and flavonoids (Zucchi et al, 2005). Different pharmacologic activities had been noticed for ginseng that depends on dose and duration. The most phytochemical and pharmacologically active compound in ginseng root is panaxosides, which had anticonvulsant, analgaesic, antipsychotic effects and stress-ulcer preventing action. Oral administration of ginseng had the capability for attenuating serum cholesterol and triglycerides, decreasing platelet adhesiveness, impair coagulation, and augmenting fibrinolysis in cholesterol- fed rats (Ibrahim, 2009). Ginseng has role in maintaining body homeostasis, improving body strength, as remedies for aging, cardiovascular disorders, immune dysfunctions, and cancer (Lee et al., 2012). So, this study aimed to investigate the hepatoprotective effect of ginseng against dexamethasone-induced side effects.2-MATERIALS AND METHODS2.1 Experimental animals:This research was conducted with strict rules to preserve and safeguard the animal welfare without subjecting them to any degree of suffering or stress. A total of 60 male rats with average B.W. of 150 g were purchased from Medical Research Institute, Alexandria University and kept for a period of 14 days for acclimatization in clean cages with a 12 hour dark/light cycle, ambient temperature of 21 ± 2.0?C and relative humidity of 44-45%. The rats were fed standard commercial chow and had free access to the drinking water.2.2 Experimental Design:All experimental procedures maintained and performed by national guidelines and protocols, approved by the?Alexandria University Institutional Animal Care and Use Committee. A total of 60 male albino rats were allocated into the equal 6 groups each group contain 10 rats as follows: group1( control): kept on balanced diet and water ad libitum; group 2: was received ginseng roots aqueous extract at a dose of 200 mg/kg B.W. / orally (Uzkeser et al., 2012); group 3 (dexamethasone treated group): was treated with dexamethasone (1mg/kg B.W. /i.p) for 7 days then slaughter (Dalatabadi and Zarchii, 2015); group 4 (dexamethasone- withdrawal group): was administrated with dexamethasone (1mg/kg B.W./i.p) for 7 days, then left untreated for further 3weeks for studying the possibility of recovery; group 5 (dexamethasone ginseng treated group) was kept on 1 week dexamethasone (1mg/kg B.W./i.p) + 3weeks ginseng (200 mg/kg B.W. orally) (Ibrahim, 2009); group 6 (ginseng-dexamethasone-ginseng group): was received ginseng for first 2 weeks then dexamethasone for one week then ginseng again for last week with same previous doses and routes.2.3 Chemicals and reagentsDexamethasone (Epideron?) was purchased from EPICO, Egypt. Panax ginseng roots aqueous extract were obtained from LETAOTAO Company China. malondialdehyde (MDA) and reduced glutathione (GSH) kits were supported by Biodiagnostic Company, Egypt. All other high grade kits and chemicals were purchased from Sigma Aldrich Co, USA.2.4 Blood collection:After 4 weeks, rats were left overnight fasted. Blood samples were collected from the retro-orbital plexus under anesthesia, in dry tubes and left for 30 minutes to be clotted at room temperature. The tubes were centrifuged at 3000 rpm for 10 minutes and the clear sera were separated then transferred into clean dry eppindorfs and kept frozen at -20?C until be used.2.5 Tissue preparation: Livers were divided into three parts; one part was immediately kept in -80?C for molecular examinations, second and third parts were washed by saline, dried on filter paper then 2nd one was kept at formalin 10% for histological studies while the 3rd one was homogenized in ice-cooled saline using mortar and Teflon homogenizer to prepare 25% w/v homogenate. Then, the homogenate was aliquot into two parts; the first one was deproteinized with ice-cold 12% trichloroacetic acid, centrifugedat 1000 xg, and the obtained supernatant was used for the estimation of reduced glutathione (GSH) content. The second part was centrifuged at 1000 xg and the resultant supernatant was used for measurement of malondialdehyde (MDA) level.2.6 Assessment of biochemical parameters:Determination of serum total cholesterol (TC) (mg/dl) according to Allain et al.,(1974), triacylglycerols (TAG) (mg/dl) according to Stein, (1987), high density lipoprotein cholesterol (HDL-c) (mg/dl) according to Burstein et al.,(1980), and calculation of low density lipoprotein cholesterol (LDL-c) (mg/dl) and very low density lipoprotein cholesterol (vLDL- c) (mg/dl) according to Bauer, (1982). Malondialdehyde, which react with thiobarbituric acid reactive substances (TBARS) (nmol/g wet tissue), was also evaluated according to method described by Satoh et al., (1987). In addition, reduced glutathione (GSH) (?mol/g wet tissue) was measured as described by Beutler et al., (1963). Serum total proteins (g/dl) was performed according to Doumas et al., (1981), albumin (g/dl) according to Doumas et al., (1971), Alanine amino transferase-Aspartate amino transferase (ALT- AST) (U/L) according to Young, (1990), alkaline phosphatase (ALP) (U/L) according to Tietz et al., (1983) and gamma glutamyl transferase (GGT) (U/L) according to Moss et al., (1987).2.7 Gene expression determination of bax and thioredoxine using mRNA extraction and reverse transcription-polymerase chain reaction?( RT-PCR):Liver tissue was used for disconnected Total RNA by using TRIZOL (GeneZOl TM RNA extraction reagent) (Intron, korea company), following the manufacturer’s protocol. Dissolving the RNA pellet in RNase free water. The concentration of RNA was measured by using Nanodrop spectrophotometer (Nanodrop 2000c, Thermo Scientific, USA). Single strand Complementary DNA was synthetized by using HiSenScriptTM[-] cDNA synthesis kit (INTRON Biotechnology, company). It was incorporated by mix 10?l of 2 X RT Reaction solutions, 1?l of enzyme mix solution, 1?g of RNA and completed by RNas free water to 20?l total volume. Incubated at 50 ?C for 30 minutes and 85?C for 10 minutes. Expressions of genes in liver tissue were analyzed by using real-time PCR. The primers were designed by (macrogen Co). All PCR reactions were done by utilizing SYBR Green qPCR Master Mix and using a strip of PCR tube (GEnetix Biotech Asia Pvt.ltd.): introductory denaturation at 92 °C for 10 minutes, followed by 40 cycles of 15 seconds at 92 °C, 30 seconds at 60 °C and 30 seconds at 72 °C. The primer sequences of all target and reference genes were recorded in Table (1). The differences in gene expression among the different groups were estimated by using the △△Ct method, standardized to β-actin and expressed as relative mRNA levels contrasted with the control.2.8 Histopathological examination: Instantly after sacrificing, one part of the liver samples was quickly preserved in 10% buffered formalin for no less than 48 hours. Thereafter, the settled examples were transformed through paraffin embedding method and were stained with hematoxylin and eosin (H&E) according to the method described by Culling, (1983).2.9 Statistical analysisAll variables were first tested for normality of data and homogeneity of group variances. Variables that lacked the assumptions of parametric methods were assessed with Kruskal-Wallis test and Dunn’s post-hoc test for multiple comparisons. Otherwise, one-way analysis of variance (ANOVA) for group effect with Boferroni post-test was used. Data are presented as mean ± standard error of mean. Probability less than or equal to 0.05 was considered significant. We used SAS software (v9.1, SAS Institute Inc., Cary, NC, USA) to run ANOVA, and SPSS (Version22.0:SPSS Inc., Chicago, Illinois, USA) to perform the non-parameteric alternatives.. 3-RESULTS3.1 Serum lipid profile.Result in Table (2) clarified that rats administrated with ginseng (G2) showed improvement of lipid profile over control one (G1). Dexamethasone treated rats (G 3, 4) had a drastic increase in lipid profile (TC, TG, LDL-c, vLDL-c) as compared with control rats. However, rats in withdrawal group (G4) had improved lipid profile and as compared with (G3). On the other hand ginseng (either for protection or therapy) successfully alleviated previous alterations as it significantly decreased (TG, TC, vLDL-C, LDL-C) while increased HDL-c in both group (5,6) as compared with untreated dexamethasone rats in addition rats that had taken ginseng before and after showed the best results. 3.2 Hepatic oxidant/antioxidant indices.Data in Table (3) demonstrated that rats administrated with ginseng (G2) showed significant increase in reduced glutathione and significant decrease in lipid peroxidation as compared with controls one (G1).Dexamethasone treated rats had marked elevation in MDA level and marked decrease in reduced glutathione as compared with control group. However, dexamethasone withdrawal group (G4) had lower lipid peroxidation and increase reduced glutathione activities as compared with (G3). On the other hand ginseng treated rats (Gs 5, 6) showed decreased lipid peroxidation and increased GSH concentrations as compared with untreated dexamethasone rats. Moreover, rats in (G6) showed rebalanced oxidant-antioxidant status. 3.3 Hepatic mRNA expression OF Bax and thioredoxin genes.Table (4) revealed that, dexamethasone treated rats had marked elevation in expression of Bax gene and marked decrease in thioredoxine expression as compared with control rats. However, dexamethasone withdrawal group (G4) had significant decrease in Bax gene expression and non-significant increase in thioredoxine gene as compared with (G3). While ginseng treated rats (Gs 5, 6) effectively decreased Bax gene expression and significantly increased thioredoxine gene expression as compared with untreated dexamethasone rats. Moreover, rats in (G6) showed the lowest expression of Bax gene and highest expression of thioredoxin gene. 3.4 Serum liver biomarkers.Data in Tables (5, 6) cleared that rats administrated with ginseng (G2) showed improvement of liver enzyme activities (ALT, AST, ALP and GGT) and serum level of protein over controls one (G1). dexamethasone treated rats (Gs 3,4) had marked elevation in liver enzyme activities (AST, ALT,ALP, GGT) and marked decreased in total protein, albumin and globulin levels as compared with control rats. However, dexamethasone withdrawal group (G4) had lower elevated enzyme activities and lower decline in protein, albumin, and globulin level as compared with (G3). On the other hand ginseng treated rats (5,6) showed improvement in liver enzymes activities as their activities were significantly decrease and protein, albumin, globulin levels were significantly increased as compared with untreated dexamethasone rats moreover, rats in(G 6) showed the greatest improvement.3.5 Histopathological findings. The histological section of liver of control animal showing normal hepatocytes arranged in cords around the central vein (arrow), Fig (1a). Liver of ginseng treated animal showing normal hepatocytes arranged in cords around the central vein (arrow), Fig (1b). Liver section of dexamethasone-treatedanimal showing marked diffuse vacuolar and hydropic degeneration of the most of the hepatocytes (arrows) and focal coagulative necrosis of some hepatocytes (A) Fig (1c). Liver of dexamethasone-treated animal (withdrawal group) showing mild to moderate hepatic vacuolation and periportal mononuclear infiltration of mononuclear cells (arrow) Fig (1d). Liver of pre dexamethasone treated with ginseng showing marked decrease of hepatic vacuolation (arrow), Fig (1e). Liver of pre and post- dexamethasone treated with ginseng showingnormal hepatocytes (arrows) with presence of regenerative hepatocytes (arrowheads) Fig (1f).4-DISCUSSIONDexamethasone is a synthetic glucocorticoid used for treating and prevention of many inflammatory and autoimmune diseases. However, it could be associated with a wide range of side effects, including liver and kidney dysfunction (Duma et al., 2006). The current study high light on the possible mechanisms of hepatoprotective role of ginseng. Dexamethasone treatment caused significant increase in lipid profile which may be attributed to the catabolic effect of dexamethasone to supply high demand of energy during induced stress condition to be used as an alternative source of energy causing high TG level (Akinloye et al., 2011) and this came in accordance with Bhujbal et al., (2012) who recorded excess mobilization of fat from the adipose tissue, increase vLDL secretions by liver, decreasing hepatic lipoprotein lipase activity that inhibit the removal of plasma vLDL during dexamethasone treatment. In addition, Lingaiah et al., (2012) mentioned that high cholesterol level may be explained by reducing concentration of LDL-c receptors on hepatocyte resulted in higher LDL-c level. Dexamethasone may also interfere with monocytes function in removing excess cholesterol as well as inhibition of cholesteryl esters hydrolysis. While treatment with Ginseng showed hypolipidemic activity as it alleviated stress induced by dexamethasone, reducing energy demand and lowered cholesterol owing to its active principle (Ginsenosides) that known as powerful hypocholesterolemic agent through increasing LDL receptors synthesis in rats Kwak et al., (2010), Al Jaff and Al-Shawi., (2015). Along with hyperlipidemic condition induced by dexamethasone there was a significant increase in liver lipid peroxidation (MDA) and a significant decrease in reduced glutathione levels. These results were compatible with those obtained by Whitworth et al., (2001) Who mentioned that oxidative stress might be pronounced with prolonged glucocorticoid exposure leading to numerous deleterious consequences for the cell, including cell death which was matched with our finding regarding up regulation of Bax gene which also came in accordance with findings of Amaral et al., (2009) who stated that glucocorticoids (GCs) evoke apoptosis through regulating BCL2 Associated X, Apoptosis Regulator (Bax) mRNA and proteins expression after activating glucocorticoids receptor. The synthetic glucocorticoid (GC), dexamethasone was found to induce changes in mitochondrial membrane properties, reduce expression of mitochondrial transporters of substrates and proteins, which lead to repressed mitochondrial respiratory activity and lower cellular Adenosine triphosphate (ATP) levels, which contributed to apoptosis Eberhart et al., (2011). Reduction in the activity of total antioxidant enzymes might be due to their utilization by enhanced free radical production during dexamethasone metabolism Lingaiah et al., (2012). This goes in accordance with our finding regarding down regulation of Thiroredoxin gene expression. However, Ginseng showed antioxidant activities as it contains ginsenosides, phenolic acids, flavonoids, and saponin which are known as powerful anti-oxidant agents enabling it to reduce oxidative stress induced by dexamethason and improving antioxidant status along with regulation of Bax and Thiroredoxin genes expression. Moreover, He et al., (2012) recorded that Panax ginseng had regulatory role of apoptosis through modulation expression pattern of certain key genes BCL2 Associated X, Apoptosis Regulator and B-cell lymphoma-extra large?(Bcl-2, Bcl-xL). As a result of previous hyperlipidemic condition and oxidative stress existence dexamethason treated rats there was pronounced induced liver injury at the end of the experimental period which came in accordance with Pierre Kamtchouing et al., (2014) who demonstrated that administration of dexamethasone induce a significant increase of AST, ALT, ALP, and GGT activities and bilirubin level that indicates damage to cell and thus injury to liver. It is important to point out that, GGT activity is one of the best indicator of liver damage as reported by Hussain et al., (2012). Also, dexamethasone treatment resulted in marked decrease of serum total protein, albumin as compared with group 1 that may resulted from liver damage which accompanied by increased rate of catabolism rather than impairment of synthesis Moreover, free radicals during dexamethasone metabolism are capable of damaging biological molecules such as proteins that have an impact on cell activities as well as membrane functions and structure. However, pretreatment with Panax ginseng, modulated the values of AST, ALT, and albumin significantly towards the normal values, as it decreased liver damage and protect hepatocyte from toxicants as they reduce entry of toxicants to hepatocyte, maintaining integrity of membrane and scavenging free radicals that cause instability of hepatocyte membrane there by suppressing leakage of enzymes Kim (2016). Protection with ginseng increased serum protein, albumin as they have regeneration ability to hepatocyte and ginseng rich with flavonoids and polyphenols which are powerful antioxidant that act as scavengers of free radicals that cause damage to protein and albumin. Ginsenosides have also been reported to stimulate ribonucleic acid (RNA) transcription and protein synthesis decreasing apoptosis Delui et al., (2013). 5-CONCLUSIONThe obtained data revealed that, aqueous extract of ginseng showed promising protective and therapeutic properties against induced dexamethasone damages in rats as it successfully improved altered liver functions, rebalanced anti-oxidative status and reduce apoptosis so, we recommend drinking aqueous extract of ginseng concurrently with dexamethasone therapy.6-REFERANCESAkinloye, O. A.,? Adamson, O., Ademuyiwa,??T. A. 2011.?Supplementation of vitamins C, E and its combination on paraquat-intoxicated rats: effects on some biochemical and markers of oxidative stress parameters. J.?App.?Pharm.?Sci.10 (06): 85-91.Allain, C.C., Poon, C. S., Clau, C. S., Chan, W. 1974. Enzymatic determination of total serum cholesterol. Clin. Chem. 20.470-475.Al Jaff, A. H., and Al-Shawi, N. N. 2015. Effect of two doses of Panax Ginseng on serum lipid profile in both normo and hypercholesterolemic rabbits .I. J.C.P. 01 (02) : 0976-8157.Amaral, J. D., Solá, C. J., Steer, C. M. 2009. Role of nuclear steroidreceptors in apoptosis.?J. Curr.Med.Chem. 16. (29). 3886–3902.??Bauer, J. D. Clinical laboratory methods. 9th ed, the C.V Company, Westline Industrial Missouri, 1982, 63116: 533:555.Beutler, E. O., Duron, Kellin, M. B. 1963. Improved method for the determination of bloodglutathione. J. Lab. Clin. Med. 61: 882-888. Bhujbal, S. S., Providencia. C. A., Nanda, R. K., Hadawale, S. S., Yeola, R. R. 2012. Effect of Woodfordiafruticosa on dexamethasone induced insulin resistance in mice. Rev. Bras. Farmacogn. 22: 611-617.Burstein, M. H., Schoinnick, Morfin, R. 1980.Severe combined hyperlipidemia J. clin. Lab. Invest. 40: 557-560.Culling, C.F. .Handbook of histopathological and histochemical techniques. 3rd ed., Butterworth, London, 1983.Delui, M. H., Fatehi, H., Manavifar, M., Amini, M., Ghayour-Mobarhan, M., Zahedi, G. F. 2013. The Effects of Panax Ginseng on Lipid Profile, Prooxidant: Antioxidant Status and Highsensitivity C Reactive Protein Levels in Hyperlipidemic Patients in Iran, Int. J. Prev. Med. 4(9): 1045-1051.Dolatabadi, A., and Zarchii, S. 2015. The effect of prescription of different Dexamethasone doses on reproductive system. Biomed. Res. 26. (4): 656-660.Doumas, B. T. 1971. Albumin standards and the measurement of serum albuminwithbromcresol green. J. Clinica. Chemica. Acta.3 1: 87- 96.Doumas, B. T., Bayse, D. D., Carter, R. J. 1981. Candidate reference method for determination of total protein in serum. I. Development and validation. II. Tests for transferability. Clin. Chem. 27: 1642-1654.Duma, D. C. M., Jewell, J. A., Cidlovski, J. A. 2006. Multiple glucocorticoids receptor isoforms and mechanisms of post – translational modification. J. Steroid. Biochem. Mol. Biol. 102:11–21.HYPERLINK "" \l "!"Eberhart, K. J., Rainer, D. Bindreither, I., Ritter, E., Gnaigerd, R., Kofler, J. P. 2011. Glucocorticoid-induced alterations in mitochondrial membrane properties and respiration in childhood acute lymphoblastic leukemia. Biochim. Biophysic . Acta. 1807: 719–725.He, H. J., Xu, Y., Xu, C., Zhang, H., Wang, Y., He, T., Yuan, A. 2012. Cardioprotective effects of saponins from Panaxjaponicus on acute myocardial ischemia against oxidative stress-triggered damage and cardiac cell death in rats. J. Ethnopharmacol. 140: 73-82.Hussain, T. H., Siddiqui, S., Fareed, M., Vijayakumar, C.V. 2012. Evaluation of chemopreventive effect of Fumariaindica against N-nitrosodiethylamine and CCl4 induced hepatocellular carcinoma in wistarrats.Asian Pac .J. Trop. Med. 3(4): 623-629.Ibrahim, D. A. 2009. Study of the Effect of Panax Ginseng versus Gliclazide on Hyperglycaemia Induced by Dexamethasone in Experimental Animals.Sultan.Qaboos. Univ. Med .J. 9(1): 63–69.Kamtchouing, P. B., Nkono, L. N., Okeng, D. S., Paul. D. D. 2014. Antihyperglycemic and Antioxidant Properties of Alstoniaboonei De Wild. (Apocynaceae) Stem Bark Aqueous Extract in Dexamethasone-Induced Hyperglycemic Rats. Int. J .Diabetes?.Res. 3(3): 27-35. Kim T-W. 2016. Ginseng for Liver Injury?Medicines(Basel). 3(4): 33.Kinouchi, S. 2003. Changes in Apoptosis-Related Genes (Bcl-2, Bax) in the Urethras of Old Female Rats Following Estrogen Replacement.Yonago. Actamedica.46:109–115.Kwak, Y. S., Kyung, J. S., Kim, J. Y. Cho, Rhee, M. H. 2010. Antihyperlipidemic effects of red ginseng acidic polysaccharides from Korean red ginseng. Biol. Pharm. Bull. 33(3): 468-472.Lee, S. H., Lee, H. J., Lee, Y. H., Lee, B.W., Cha, B. S., Kang, E. S., Ahn, C.W., Park, J. S. 2012. Korean red ginseng (Panax ginseng) improves insulin sensitivity in high fat fed Sprague-Dawley rats. Phytother. Res. 26(1): 142-147.Lingaiah, H. B., Thamaraiselvan, R., Periyasamy, B. 2012. Dexamethasone induced alterations in lipid peroxidation, antioxidants; membrane bound ATPase in wistar albino rats. Int. J. Pharm. Pharm . Sci. 4:497-449.Mansur, K.Y., Iwahashi, S. Kiryu-Seo, Q. Su, K., Namikawa, J. Yodoi, Kiyama, H. 1998. Brain Research. Mol. Brain .Res. 62: 86-91.Moss, D.W., Henderson, A. R., Kachmar, J. F. 1987. Enzymes. In: Tietz, NW, ed. Fundamentals of clinical chemistry. 3 rd ed. Philadelphia: WB Saunders; 1987, 346-421.Satoh, K. 1987. Serum lipid peroxide in cerebrovascular disorders determined by a new colorimetric method. Clin. Chem. Acta. 90: 37-42 Stein, E. A. Lipids, lipoproteins.Tietz NW. Fundementals of clinic chem. 3ed:.Philadelphia: WB Saunders; 1987, 448-481.Tietz, N. W., Rinker, A., Shaw, L. M. 1983. IFCC methods for the measurement of catalyticconcentration of enzymes.Part 5.IFCC method for alkaline phosphatase. J. Clin. Chem. Clin. Biochem. 21: 731-748Uzkeser, M. E, Karakus, A., Albayrak, A., Kiki, I., Bayir, Y., Cadirci, E., Unal, U., Halic, Z., Karadeniz, A. 2012. Protective effect of Panax ginseng against Nacetylpaminophenol-induced hepatotoxicity in rats.African J. Pharm. Pharmacol. 6(36): 2634- 2642.Wentzel, P. M., G?reskog, Eriksson, U. J. 2008. Decreased Cardiac Glutathione Peroxidase Levels and Enhanced Mandibular Apoptosis in Malformed Embryos of Diabetic Rats. Diabetes. 57(12): 3344–3352.Whitworth, J.A., Schyvens, G.G., Zhang, Y., Mangos, G.I., Kelly, J.J. 2001.Glucocorticoid-induced hypertension: from mouse to man. Clin. Exp. Pharmacol. Physiol. 28: 993–999.Young, D. S. 1990. Effect of drugs on clinical laboratory tests. Physiol. Res. 59: 61-70.Zucchi, O. L. Moreira, S., de Jesus, E. F., Neto, H. S., Salvador, M. J. 2005. Characterization of hypoglycemiant plants by total reflection X-ray fluorescence spectrometry.?Biol Trace Elem Res. 103: 277–290. ................
................

In order to avoid copyright disputes, this page is only a partial summary.

Google Online Preview   Download