Abstract is an essential trace element and ha been shown ot protect the rats against dietary liver necrosis.

This study was designed to evaluate the effects of selenium supplementation on different biochemical parameters in thioacetamide induce cirrhotic rats, for this purpose 24 ,a;e A;nomp wostar rats were dvided into four groups (n=6). Group I remain healthy control rats group II, received thioacetamide at a dose of (2amg/kg b.w, i.p, for 12 weeks, twice a week) Group III, received thioacetamide (200mg/kg b.w, i.p for 12 weeks, twice a week) and then sodium selenite (1mg/kg b.w, i.p for 12 weeks, three times a week, twice a week) and then sodium selenite (1mg/kg b.w k.p for 12 weeks , three times a week). Biochemical analysisi was evaluated by Total and direct bilirubin, liver specific enzymes, and antioxidant enzumes marked increase in ttotal and direct bierubin and ALLT activity was the indicative markers of liver cirrhosis while reduced antioxidant. Activity (SOD and GSH) and increased MDA and catalase levels were observed in cirrhotic group. Sodium selenite supplemetatio markedly redced totoa bilirubin and ALT activity and restored the antioxidant enzymes (SOD and GSH) and MDA and catalase activity. These results indicate that sodium selenit successively attenuates the thioacetamide induce liver cirrhosis. Key words: Sodium selentie, Thioacetamide, liver enzymes, SOD, GSH, catalase MDA. Introduction: Liver cirrhosis is characterized by an abnormal accumulation of fibrous tissue and by ehpatocytes regeneration. [1] The mechanism of liver cirrhosis have been investigated using several animal models such as carbon tetrachloride treatment [2,3]. Bile duct ligation [4], alcoholism [5] and thioacetamide administration [6,7]. It has been reported that liver fibrosis as well as regenerative hepatic nodiles were more prominent in TAA

induced cirrhosis models than in CCl4 treated rats and that the pathology of the TAA included cirrhosis was more similar to that of human cirrhosis [8]. Thio acetamide, whih was or Iginal used as a fungicide is a hepatoxin commonl ysed to induce liver cirrhosis in rats [9]. Biotranformation of TAA precedes oxidative damage associated liver injury [10]. This has been implicated earlier by the presence of glutathione depletion [11], an increase in malondialdehyde [12], reduction in superoxide dismutase [13,14] and increase om catalase activity [15] in liver cells following TAA administration. The levels of selenium, a naturally occurring micronutrient, were significantly reduced in TAA treated rats [16]. Supplementation with selenium reversed the changes induced by thioacetamide [17]. The mode of action of selenium is unkonown but may involve antioxidant defence mechanism [18]. Its seleno enzyme glutathione peroxidase (GSHPX) protects the biomembrances fro m oxidative edestruction. This enzyme is present n I many tissues of the body, and the decreased level of this enzyme increases the tendency of cellular damage. Glutathione peroxidase is depressed in selenium deficient animals [20,21]. Selenium , along with other antioxidants such as Vitamin E and superoxdie dismuatase protects the cell lagaints oxidant damage. [22,23]. In views of above mentioned previous studies it is hypothised that cirrhosis of the liver could be prevented by the use of some antioxidants. The present study was undertaken to examine the underlying mechanism of effect of selenium treatment on different biochemical parameters in experimental rats model. Materials and methods: 24 male Albino Wistar rats weighing 200-250gm were purchased from the animal house of ICCBS (International center for chemical and biological sciences, Karachi, Pakistan) for the study. Animals were acclimatized to the laboratory

conditions before the start of experiment and caged ina quiet temperatutre controlled animal room (23+4oC). Rats haed free access to water and standard rat diet the experiments were conducted in accordance with ethical guidelines for investigations in laboratory animals. Study design: The rats were randomly divided into four groups each of six rats. The duration of the study was 26 weeks, divided into two phase thioacetamide and sodium selenium (Na2SeO3) were administered in either phase. Group I was the control and reminded untreated. Phase I: In Phase I, group II and III received TAA, dissolved in 0.9% NaCl and was injected intraperitoneally at a dosage of 200mg/kg, b.w, twice a week for 12 weeks. Group IV received sodium selenite intrapertoneally at a dosage of 1mg/kg b.w, three times a week for 12 weeks. During the 13th week the animals were given standard laboratory diet and water adlibitum. At the end of 13th week, the animals of group I (control), group II (received TAA) and group IV (received TAA) group IV receive sodium selenite were decapitated. Phase II: In Phase II, the animals of group III were given sodium selenite intraperitoneally at a dosage of 1mg/kg b.w, 7 three times a week, starting from 13th week for 12 weeks. During the 26th week, animals were given standard laboratory diet and water adlibitum. At the end of the experimental period the animals were anesthetized, decapitaled and the blood was sampled from the head

wound in the lithium heparinized coateded tubes. The sampewere stored at 70oC. A portion of blood was taken in separate tube to collect the plasma. ASSESSMENT OF LIVER ENZYMES: Plasma ALT (Rietman and Franhel, 1957) total and direct bilirubin (Sherlock 1951) were analyzed using commercially prepared reagent kits from Randox. Assessment of tissue superoxide dismultase. Tissue superoxide dismutase was estimated spectrophotometrically by the method of Kono. (Kono et al 1978) Assessment of tissue malondialdehy de. Tissue MDA was estimated spectrophotometrically by the lipidperoxidation method (ohkawa et al. 1979) Assessment of tissue catalase. Tissue catalase was estimated spectrophotometrially by the method of sinha Sinha 1972) Assessment of tissue Glutathione Reductase. Statistical analysis: Results are presented as mean + SD . Statistical significance and difference from control and test values evaluated by students t-test statistical probability of P<0.01, P<0.001 were considered to be significant.

Result: Supernatant so obtained was centrifuged at 10, 500g for 20 minutes at 4oC to get post mitochondrial supernatant. Which was used to assay superoxide dismutas [24], catalas [25], malondiadehyde [26] and glutathione reductase [27] activities. Plasma samples were used to assay Alanine aminotransferase [28] and total and direct bilirubin levels [29]. Statistical analysis: Results are presented as mean + SD Statistical significance and difference from control and test values evaluated by students t-test. Statistical probability of P<0.01, P<0.001 were considered to be significant. Results: Effects of Thioacetamide and sodium selenite treatment on body weight in control and treated animals. Decreased body weight was observed after chronic administration of TAA in group II and group III animals. Group III animals regained their body weight after sodium selenite treatment in second phase. (Fig-1) Effects of thioacetamide and sodium selenite treatment on plasma. Liver enzymes in control and treated rats: Table I shows a marked increase in total bilirubin level in group II animals as compared to control (0.31+0.01 P<0.001) where as, in group III an imals, sodium selenite supplementation brought those increased levels as the normal concentrations as compare to control (0.12+0.002, P<0.01). Increased levels of direct bilirubin was shown by group II animals as compare to control (0.82+0.03,

P<0.001) where as sodium selenite supplementation brought those higher levels to normal levels as compare to control but result was not significant. Plasma Alanine aminotransfers level was markedly increased in group II animals as compare to control (40.13+2.0, <0.001). Alanine amino transferase Level was decreased in group III animals as compare to control (8.7+0.18, P<0.001). Effects of thioacetamide and sodium selenite treatment on plasma liver enzymes in control and treated rats. Parameters Total bilirubin (unit/L) Direct bilirubin (Unit/L) Alanin transferase Group I 0.16+0.01 Group II Group III 0.31+0.002 0.12+0.002 Group IV 0.39+0.001 0.20+0.01 10.24+0.01

0.052+0.03 0.82+0.001 0.040+0.001 6.56+0.18 40.13+0.18 8.7+0.18

(Unit/L) Values are mean + SD. Significant difference among control, Thioacetamide and sodium selenite treated groups by t-test P<0.01, P<0.001. Effects of thioacetamide and sodium selenite treatment on hepatic concentration of Glutathione Reductase. Hepatic concentration of glutathione reductase was significantly reduced in

group II animals as compare to control (0.026+0.001, P<0.001). Animals of group III, after sodium selenite supplementation, showed as normal levels of glutathione reductase to control (0.5+0.01) Table 2. (Glutathione reductase was increased in group IV as compare to control (1.7+0.03, P<0.001) effects of thioacetamide and sodium selenite treatment on hepatic concentration of MDA. Level of MDA was markedly increased in group II animals as compare to control.

Sodium Selenite treatment (Table-2). Sodium selenite administration in group IV decreased the concentration of MDA as compare to control (7.81+0.001) Table-2: Effects of thioacetamide and sodium selenite treatment on hepatic concentration of Glutathione Reductase, superoxide dismutase, Malondialdehyde and catalase. Parameters Glutathione Reductase (unit/gm of 17.0+0.3 53.26+1.3 100.03+5.9 15.3+0.01 282.8+4.0 7.81+0.01 tissue) SOD unit/gm 379+20.0 of tissue. MDA mol/gm tissue Catalase nmol/gm tissue. Values are mean + SD. Significant difference among control, thioacetamide, thioacetamide and sodium selenite and sodium selenite treated groups by t-test. P<0.01, P<0.001. Effect of thioacetamide and sodium selenite treatrment on hepatic concentration superoxide dismutase in control and treated rats. Table-2: showed a significant decrease in SOD activity in group II as compare to control (379+20.0). Animals of group III, after sodium eslenite suopplementation, of n 21.7+2.8 of 8.63+0.02 38.91+0.12 10.68+0.16 11.98+0.16 Group I 0.64+0.02 Group II 0.026+0.001 Group III 0.5+0.01 Group IV 1.7+0.03

showed normal levels of SOD activity (100.03+5.9,P<0.001) as compare to control. SOD activity was normal in group IV animals (282.8+4.0, P<0.001) as compare to control. Effect of thioacetamide and sodium selenite treatment on hepatic concentration of catalase. Concentration of catalase was significantly increased in group II animals (38.91+0.12, P<0.001) as compare to control. Administration of sodium selenite in second phase in group III animals brought these highe5 levels to normal limits (10.68+0.16, P<0.001) as compare to control. Level of catalase was normal (11.98+0.16, P<0.001) in group IV as compare to control (Table -2).