北豆根致大鼠肝损伤血清酶生物标志物的早期变化及联合检测

摘要

目的：探讨总胆汁酸（TBA）、α-谷胱甘肽S转移酶（α-GST）等血清酶生物标志物对北豆根致大鼠肝损伤早期诊断的应用价值。方法：Wistar大鼠随机分为对照组和北豆根低、高剂量组（22.5、45 g · kg-1），每组18只。给药组每日灌胃给药1次，连续21 d，对照组灌服等体积蒸馏水。分别于给药7 d、14 d、21 d处理动物，每组6只，收集血清，采用全自动生化仪测定血清丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）、碱性磷酸酶（ALP）、总胆红素（TB）以及TBA的水平，采用酶联免疫法（ ELISA）测定血清α-GST、嘌呤核苷磷酸化酶（ PNP）、鸟氨酸氨基甲酰转移酶（ OCT）和精氨酸酶Ⅰ（ Arg Ⅰ）的水平；计算大鼠的肝脏指数及观察肝组织病理学变化；应用Logistic回归和ROC曲线评价上述指标对北豆根致大鼠肝损伤早期诊断的意义。结果：与同时间点对照组相比，高剂量组给药14 d ALP升高（P＜0.05），14 d、21 d ALT和肝脏指数升高（P＜0.05），组内有2∕6肝组织出现肝细胞点状坏死、小灶性肝细胞坏死和炎症细胞浸润；北豆根低剂量组肝脏指数于给药21 d升高（ P＜0.01）。北豆根高剂量组给药7 d、14 d、21 d血清α-GST、PNP和ArgⅠ升高（P＜0.05或P＜0.01），OCT 14 d、21 d升高（P＜0.01），TBA 14 d升高（P＜0.05）。动物血清中ALT、AST、α-GST、PNP和ArgⅠ各项的ROC曲线下面积（AUC）分别为0.787、0.483、0.957、0.948和0.934。其中 ALT和 AST 联合检测的 AUC 为0.809，α-GST、Arg Ⅰ及与 PNP 联合检测的 AUC 为1.000，均高于各指标单项检测的AUC。结论：血清α-GST、PNP和ArgⅠ可作为北豆根致肝损伤早期的生物标志物，且联合检测在北豆根致肝损伤早期有较高的诊断价值。%Objective:To evaluate the application value of serum enzyme biomarkers of total bile acid ( TBA) ,α-glutathione-S-transfer-ase (α-GST) etc for the early diagnosis of liver injury induced by Rhizoma Menispermi ( RM) in rats.Methods:Wistar rats were ran-domly divided into control group and RM low dose group (22.5 g· kg-1) and high dose (45 g· kg-1) group (n=18).RM 22.5 and 45 g· kg-1 groups were treated with RM decoction by intragastrical administration , while the control group was given distilled water of the same volume once a day for 21 consecutive days.Blood was collected respectively 7, 14, 21 days after RM administration.The lev-els of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total bilirubin (TB) and TBA were measured by using automatic biochemical analyzer .The concentration of α-GST, purine nucleoside phosphorlyase ( PNP) , ornithine carbamyl transferase ( OCT ) and arginase Ⅰ ( Arg Ⅰ) were detected with enzyme-linked immunosorbent assay ( ELISA) .Liver weight was measured and the pathological changes of liver tissue were observed .Logistic regression and ROC curve were applied to analyze the data of above indexes for evaluating the early diagnostic value of liver injury induced by RM .Results:Com-pared with that in the control group at the same time point , on day 14 the level of ALP was elevated (P<0.05) and the level of ALT and liver weight index were increased (P<0.05) and some spotty and focal necrosis , inflammatory cell infiltration in the liver tissue were demonstrated on day 14 and 21 in RM high dose group.The liver weight index was increased in RM low dose group (P<0.01). The levels of α-GST, PNP and OCT were elevated on day 7, 14 and 21 (P<0.05 orP<0.01), and OCT increased on day 14 and 21, TBA increased only on day 14 in RM high dose group.The areas under the ROC curve (AUC) for serum ALT, AST,α-GST, PNP and Arg Ⅰwere 0.787, 0.483, 0.957, 0.948 and 0.934 respectively.The AUC of the combined examination of ALT and AST was 0. 809.The AUC of the combined examination of α-GST, Arg Ⅰ and the AUC of the combined examination of α-GST, Arg Ⅰand PNP were the same of 1.000, higher than the AUC of single index examination .Conclusion:α-GST, PNP and ArgⅠmay be used as the bi-omarkers at the early stage of hepatic injury induced by RM .The combined examination has a higher value for the early liver injury diag-nosis.