山豆根水煎液致大鼠亚急性肝脏毒性研究

Rat Subacute Hepatotoxicity Induced by Sophorae Tonkinensis Radix et Rhizoma

目的:给予质量可控的山豆根(RRST)水煎液致大鼠肝脏毒性,并探讨其毒性作用机制。方法:采用代表性好、来源固定的药材,取山豆根饮片加6倍量水,浸泡30 min,武火煮沸,换文火煎煮60 min,共煎煮2次,合并2次的滤液浓缩至生药2 g·mL-1,得山豆根水煎液储备液,并对其苦参碱及氧化苦参碱进行测定,对其质量进行控制。动物实验:选用Wistar大鼠30只,随机分为空白对照组(灌胃给予蒸馏水)、阳性对照组(灌胃给予10%CCl4)、低剂量组(灌胃给予1 g·kg-1山豆根水煎液)、中剂量组(灌胃给予4 g·kg-1山豆根水煎液)和高剂量组(灌胃给予16 g·kg-1山豆根水煎液),每组6只。连续给药14d,第15天ip 25%乌拉坦麻醉,腹主动脉取血,离心取血清测定丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、总胆红素(TBil)、总胆汁酸(TBA),采用酶联免疫吸附法(ELISA)法测定炎症因子白介素-6(IL-6)、白介素-10(IL-10)、肿瘤坏死因子(TNF-α)。摘取肝脏,称重,计算脏器指数,病理制片,进行组织病理学检查。取部分肝组织加冷生理盐水制成10%肝匀浆,离心,取上清液,测定肝组织中的超氧化物歧化酶(SOD)、谷胱甘肽(GSH)活性,丙二醛(MDA)含量。结果:大鼠连续给予不同剂量山豆根水煎液7 d后,高剂量组出现活动减少,四肢无力;14 d后,与空白对照组相比,高剂量组ALT,ALP,TBA值升高(P<0.05),病理组织学检查发现高剂量组的肝细胞肿大,并伴有炎症浸润。高剂量组及阳性对照组的IL-6,TNF-α含量升高(P<0.05及P<0.01),SOD活性降低,MDA含量升高(P<0.05),SOD/MDA含量显著降低(P<0.01)。结论:山豆根具有肝脏毒性,给予大鼠16 g·kg-1山豆根水煎液可导致明显的肝损伤,且其损伤机制与炎症因子的作用和脂质过氧化有关,与四氯化碳的肝毒性具有一定的相似性。

Objective： To investigate the hepatic injury induced by multiple administration of controlled Sophorae Tonkinensis Radix et Rhizoma（STRR） in rats and its toxic mechanisms.Method： The quality of extract of STRR was controlled by purchasing the herbs from the same source and the extraction： 1 hour of extraction,6 times amount of water,and twice extraction.Then the extract was used in animal experiment.The rats were divided randomly into 5 groups： blank control group,the extrad-STRR low dose group（1 g·kg-1）,medium dose group（4 g·kg-1）,high dose group（16 g·kg-1） and positive control group（10% CCl 4）.The rats were given the corresponding extract by gavage for 14 days.The toxic effect and body weight of rats were observed.After last administration,the rats were anaesthetized by 25% urethane,and blood was collected from aorta abdominalis,then the liver function and tumor necrosis factor-α（TNF-α）,interleukin（IL）-6,IL-10 in serum was detected.The livers were collected and weighed,organ index was calculated.The hepatic histopathology was examined.The value of superoxide dismutase（SOD）,malanyldiadehyde（MDA）,GSH in liver was investigated.Result： After administration of STRR for 7 days,reduced locomotor activity of the rats in high dose group was found.Compared with blank control group,the level of ALT,ALP,TBA increased significantly in high dose group and positive control group（P 0.01 or P 0.05） after 14 day of administration.Hepatocyte swelling and inflammatory cell infiltrate were evident in high dose group.The organ index of liver in high dose group increased significantly.The level of IL-6,TNF-α and the content of MDA,GSH also increased significantly in high dose group.The content of SOD in liver and SOD / MDA ratio decreased significantly in high group.Conclusion： Liver injury in rats was induced after administration of large dose of STRR decoction.Oxidative stress,inflammation may be one of the hepatic toxic mechanisms,and the mechanisms is similar to the CCl 4 to some extent.