血清维生素D结合蛋白水平检测对大鼠肝毒性诊断的评价研究

Evaluation of Serum Vitamin D Binding Protein Levels in the Diagnosis of Hepatotoxicity in Rats

目的评价血清维生素D结合蛋白(vitamin D binding protein,VDBP)用于大鼠肝毒性诊断的价值。方法用酮康唑(ketoconazole,KTZ)、四氯化碳(carbon tetrachloride,CCl4)和对乙酰氨基酚(acetaminophen,APAP)在雄性大鼠中构建3种肝毒性模型,在不同时间点从腹腔大静脉采集外周血,3 000 r/min离心制备血清。用ELISA法测定血清中VDBP含量,用全自动生化分析仪测定血清中的丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)、碱性磷酸酶(alkaline phosphatase,ALP)、总胆红素(total bilirubin,TBIL)、总胆汁酸(total biliary acid,TBA)、胆碱酯酶(cholinesterase,CHE)、谷氨酸脱氢酶(glutamate dehydrogenase,GLDH)、5’核苷酸酶(5’-nucleoticlase,5’-NT)、单胺氧化酶(monoamine oxidase,MAO)和苹果酸脱氢酶(malate dehydrogenase,MDH)含量。实验组与对照组进行t检验分析,检查指标值的差异是否具有统计学意义。结果在CCl_4肝毒性模型中,与对照组相比,ALT,AST,TBIL和CHE在CCl_4给药后4 h起升高,差异均具有统计学意义(均P<0.05)。GLDH,MAO和MDH在给药后6 h起升高,差异均具有统计学意义(均P<0.05)。在KTZ肝毒性模型中,与对照组相比,ALT在KTZ给药6天后升高,差异具有统计学意义(P<0.05)。AST和TBIL在给药后8天升高,差异具有统计学意义(P<0.05)。在APAP肝毒性模型中,与对照组相比,ALP在APAP给药后4 h和8 h点升高,差异具有统计学意义(P<0.05)。CHE在给药后8 h点升高,差异具有统计学意义(P<0.05)。ALT和TBIL在给药后24 h点升高,差异具有统计学意义(P<0.05)。在3种肝毒性模型中,VDBP含量在给药后的所有时间点都降低,差异均具有统计学意义(均P<0.05)。结论 VDBP比其它肝功能指标能更灵敏地预测肝毒性的发生。

Objective To evaluate the value of serum vitamin D binding protein(VDBP) in the diagnosis of hepatotoxicity in rats.Methods Three hepatotoxicity models were constructed in male rats with ketoconazole(KTZ),carbon tetrachloride(CCl4) and acetaminophen(APAP),peripheral blood was collected from large abdominal veins at different time points and serum was prepared by 3 000 r/min centrifugation.The concentration of VDBP in serum was determined by ELISA,and the contents of alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),total bilirubin(TBIL),total biliary acid(TBA),cholinesterase(CHE),glutamate dehydrogenase(GLDH),5’-nucleoticlase(5’-NT),monoamine oxidase(MAO) and malate dehydrogenase(MDH) in serum were determined by automatic biochemical analyzer.The t test was carried out in the experimental group and the control group to check whether the differences in the indicators of each time point were statistically significant.Results In the CCl4 hepatotoxicity model,compared with control group,ALT,AST,TBIL and CHE increased from 4 h after CCl4 administration,and the difference was statistically significant(P<0.05).GLDH,MAO and MDH increased from 6 h,and the difference was statistically significant(P<0.05).In the KTZ hepatotoxicity model,compared with control group,ALT increased from 6 days after KTZ administration,and the difference was statistically significant(P<0.05).AST and TBIL increased at 8 days,and the difference was statistically significant(P<0.05).In the APAP hepatotoxicity model,compared with control group,ALP increased at 4 h and 8 h after APAP administration,and the difference was statistically significant(P<0.05).CHE increased at 8 h,and the difference was statistically significant(P<0.05).ALT and TBIL increased at 24 h,and the difference was statistically significant(P<0.05).VDBP concentration dropped at all time points after administration in three hepatotoxicity models,and the difference was statistically significant(P<0.05).Conclusion VDBP is more sensitive than other liver function indicators to predict the occurrence of hepatotoxicity.