亚硒酸钠对四氯化碳损伤草鱼肝原代细胞与肝组织的保护作用

THE ROLE OF SODIUM SELENITE IN PROTECTION OFPRIMARY HEPATOCYTES AND LIVER AGAINST CARBONTETRACHLORIDE TOXICITY IN GRASS CARP

不同浓度四氯化碳（ＣＣｌ４）对草鱼肝原代细胞的损伤实验中，ＣＣｌ４浓度为１０μｌ／ｍｌ可引起细胞血清中丙氨酸氨基转移酶（ＡＬＴ）、天门冬氨酸氨基转移酶（ＡＳＴ）、乳酸脱氢酶（ＬＤＨ）逸出量与细胞破损率显著增高，培养液中添加亚硒酸钠（Ｎａ２ＳｅＯ３）０．２μｇ／ｍｌ，则可降低ＡＬＴ、ＡＳＴ、ＬＤＨ的逸出量，减轻细胞破损程度。Ｎａ２ＳｅＯ３保护实验中，Ｎａ２ＳｅＯ２＋ＣＣｌ４组预先腹腔注射（ｉｐ）０．１ｍｇ／ｋｇ．ｂｗ连续三日，末次ｉｐＣＣｌ４混合液１ｍｌ／ｋｇ．ｂｗ，２４ｈ内肝组织超氧物歧化酶（ＳＯＤ）相对活性比ＣＣｌ４组提高达９１．５％，第七日仍提高达５４．５％，与对照组的水平基本接近；血清中丙氨酸氨基转氨酶（ＡＬＴ）水平逐渐降低。本实验还观察到Ｎａ２ＳｅＯ３可引起肝脂质过氧化物显著降低，肝微粒体蛋白含量与细胞色素Ｐ—４５０活性升高；组织切片观察显示肝组织损伤程度减轻，７２ｈ后细胞核增多。表明Ｎａ２ＳｅＯ３可提高草鱼肝清除自由基能力，增强肝脏解毒功能。

The effect of different concentrations of carbon tetrachloride(CCl4)was studied in theprimary culture of grass carp hepatocytes.As the concentration of 10μl/3×106 cells CCl4,the increase of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)as wellas lactate dehydrogenase(LDH) released all from the hepatocytes was observed,and so wasthe increase of the percentage of damaged cells. The adding of sodium selenite(0.2μg/ml)toRPMI-1640 liquid medium and cultured hepatocytes reduced the release of ALT,AST,LDHand the extent of the cell damage.In another experiment evaluating the effect of sodiumselenite and CCl4,grass carp fingerlings were injected intraperitoneally withNa2SeO3 0.1mg/kg.bw each day for three consecutive days, and then with 1ml/kg.BwCCl4.Twenty-four hours later,the relative activity of hepatic superoxide dismutase (SOD)was found to rise 91.5% higher in these fingerlings than those injected with CCl4 only.Sevendays later it was still 54.5% higher.The level of serum ALT decreased gradually.The activitylevel of hepatic lipid peroxide decreased significantly (P < 0.05) and that of P-450 increasedsignificantly(P<0.01) and so did the contained hepatic microsomal protein(P<0.05)inthose fingerlings injected with selenium and then CCl4. Light microscopic observations revealed a decrease of hepatic damage and an increase of nuclear numbers,suggesting that celldivisions might have taken place.It is indicated that sodium selenite may elevate the capacityof hepatic SOD in scavening free radicals,and hepatic counteractions on CCl4 toxicity.