茵陈五苓散对α-萘异硫氰酸酯致大鼠黄疸型肝损伤的血清代谢组学研究

Serumal metabonomic analysis of α-naphthyl isothiocyanate induced icteric liver injury rats after treatment with Yinchen Wuling powder

目的用代谢组学的方法研究α-萘异硫氰酸酯(ANIT)致大鼠黄疸型肝损伤的代谢标记物,探讨茵陈五苓散治疗黄疸型肝损伤的作用机制。方法按照体重将SD大鼠随机分为6组,每组10只:正常组、模型组、对照组和低、中、高3个剂量实验组。实验组以茵陈五苓散水煎液8.7,17.4,25.5 g·kg^(-1)·d^(-1)灌胃;对照组以熊去氧胆酸10 mg·kg^(-1)·d^(-1)灌胃;其余2组灌胃等剂量0.9%Nacl 2.0 m L·d^(-1),连续给药15 d。一次性灌胃ANIT橄榄油溶液建立肝损伤模型,48 h后取血清,用气相色谱质谱联用仪提取血清数据,结合主成分分析和叠加偏最小二乘判别分析对数据降维,比较模型组与正常组的VIP值,寻找差异性代谢物,并对其参与的代谢途径进行分析。结果共找到14个代谢标志物,模型组的14个代谢标记物的相对峰强度与正常组比较,差异均有统计学意义(P<0.05,P<0.01,P<0.001)。相对峰强度:模型组的亚油酸(1.72±0.60)%和柠檬酸(0.58±0.16)%下降,而花生四烯酸(3.07±0.80)%和丝氨酸(2.78±0.74)%上升;茵陈五苓散干预后,高剂量实验组的琥珀酸(0.43±0.23)%、柠檬酸(2.72±2.02)%、亚油酸(7.44±4.54)%、甘油酸(0.31±0.14)%、肌醇(4.40±3.70)%恢复至正常水平,高剂量实验组与正常组比较,差异无统计学意义(P>0.05)。结论茵陈五苓散对ANIT所致的黄疸型肝损伤有一定的干预作用,其机制可能与调节氧化应激、三羧酸循环和氨基酸代谢途径有关。

Objective To find out metabolic markers for hepatic injury caused by a-naphthyl isothiocyanate （ANIT） in rats with jaundice,and to explore the mechanism of Yinchen Wuling powder. Methods According to the weight, SD rats were randomly divided into 6 groups, 10 rats in each group： normal group, model group, control group and the low, mi- ddle and high doses of three experimental groups. The experimental group was treated with 8.7, 17.4, 25.5 g·kg-1·d-1 Yinehen- Wuling pow- der water decoction;control group was treated with ursodeoxycholic acid for 10 mg · kg-·d-l;the other two groups were administered with 0.9%NaCI 2.0 mL · d-1for eonsective fifteen days. The ANIT olive oil solution was given orally in single dose to establish the model. The serums were obtained after 48 h later. The serum data was extracted by gas chromatography -mass spectrometry. The principal component analy- sis and sparse partial least squares discriminant analysis were combinedto reduce the dimension of the data. The VIP values of the model group and the normal group were compared to find the different metabolites and their involved metabolic pathways were analyzed. Results Fourteen metabolic markers were identified, There were significant differences in the relative peak intensity of metabolic markers between the model group and the normal group （P 〈 0. 05,P 〈 0. 01, P 〈 0. 001 ）. The relative peak intensity： linoleic acid （1.72 ± 0. 60）%, citric acid （0.58±0.16）% in the model group decreased, and the arachidonic acid （3.07±0.80）%, serine （2.78 ± 0. 74） % increased. After intervention by Yinchen Wuling power, the succinic acid was （ 0. 43 ± 0. 23 ） %, citric acid was （2.72 ±2. 02）%, linoleic acid was（7.44 ±4.54）%, glyceric acid was（O. 31 ±0. 14）% and inositol was （4. 40 ± 3.70 ） % in high dose experimental group, which have no significant differences with those in normal group （P 〉 0. 05 ）. Conclusion The Yinchen Wuling powder has a certain treatment on the jaundice type hepatic jury induced by ANIT, its mechanism may be related to the boxylate metabolism and amino acid metabolism. regulations of oxidative stress metabolism, glyoxylate and dicar-boxylate metabolism and amino acid metabolism.