银杏酸对大鼠肝毒性的影响研究

Effect of Ginkgolic acid on hepatotoxicity in rats

目的研究银杏酸(17∶1)致肝损伤的机制,为合理安全应用银杏酸提供依据。方法 SD大鼠随机分为空白组、对照组和低、中、高剂量实验组。低、中、高剂量实验组给予20,50,100 mg·kg^(-1)银杏酸,灌胃,空白组和对照组给予等量的生理盐水。连续给药14 d,对照组末次灌胃结束后腹腔注射40%CCl_4橄榄油建立急性肝损伤模型。取肝组织,苏木素-伊红(HE)染色后,在光学显微镜下进行病理检查,脱氧核糖核苷酸末端转移酶介导的缺口末端标记(TUNEL)法检测肝组织细胞凋亡情况,按照试剂盒方法测定肝组织匀浆液中谷胱甘肽(GSH)、超氧化物歧化酶(SOD)和丙二醛(MDA)含量。结果病理学显示,低、中、高剂量实验组出现不同程度的肝损伤,以高剂量实验组最严重。与空白组相比,大鼠肝组织SOD和GSH含量降低明显降低(P<0.01)。低、中、高剂量实验组MDA水平分别为(5.99±0.80),(10.47±0.86)和(19.13±1.69)nmol·mg^(-1)·prot^(-1),与空白组的(3.43±0.37)nmol·mg^(-1)·prot^(-1)相比,差异均有统计学意义(均P<0.05)。TUNEL法显示,低、中、高剂量实验组的细胞凋亡率分别为(26.64±4.03)%,(39.38±3.72)%和(57.87±4.64)%,与空白组比较,差异均有统计学意义(均P<0.05)。结论银杏酸(17∶1)具有明显的肝毒性,其损伤途径与引起机体氧化应激后诱导脂质过氧化有关。

Objective To explore the relationship of oxidative damage mechanism on hepatic toxic injury caused by Ginkgolic acid（17∶1）in rat and provide the basis for its safe and reasonable use.Methods Rats were randomly divided into blank group（normal saline）,control group and low,middle,high dose experimental groups（20,50,100 mg·kg^-1Ginkgolic acid）.After the last administration of normal saline,chemical hepatic injury was induced by intraperitoneal injection of CCl_4in control group.The levels of malondialdehyde（MDA）,superoxide dismutase（SOD）and glutathione（GSH）in liver tissue of rats were determined,hematoxylin-eosin staining was used to observe the pathological changes in liver tissue,terminal deoxynucleotidyl transferased UTP nick-end（TUNEL）was used to detect the cell apoptosis rate.Results Pathology showed that low,middle and high dose experimental groups had different degrees of liver injury,with the highest injury in high-dose experimental group.Compared with blank group,rats in model group showed pathological changes in edema,disordered cell arrangement and local necrosis.The levels of MDA in low,middle and high dose experimental groups were（5.99±0.80）,（10.47±0.86）and（19.13±1.69）nmol·mg^-1·prot^-1,had significant difference with that in blank group,which was（3.43±0.37）nmol·mg^-1·prot^-1（all P〈0.05）.The TUNEL assay showed that the apoptotic rates in low,middle and high dose experimental groups were（26.64±4.03）%,（39.38±3.72）%and（57.87±4.64）%,all had significant difference with that in blank group（all P〈0.05）.Conclustion The continuous administration of Ginkgolic acid（17∶1）on rat can induce obvious hepatotoxicity injury,the approach of hepatic damage may be related with the peroxidative damage mechanism.