黄肝煎液对胆汁淤积性肝损伤大鼠的保护作用及急性毒性研究

Protective Effects on Cholestatic Liver Injury in Rats and Acute Toxicity of Huanggan Decoctions

目的:观察医院制剂黄肝煎液对胆汁淤积性肝损伤的保护作用及急性毒性。方法:将72只SD大鼠随机分为正常对照组、模型组、阳性对照组和黄肝煎液低、中、高剂量组,每组12只,均连续灌胃给予相应药物14天,每天2次。除正常对照组外,其它各组均于给药第12天灌胃给予α-萘异硫氰酸酯(ANIT)建立胆汁淤积性肝损伤大鼠模型。采用全自动生化分析仪检测大鼠血清肝功能指标丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、羟脯氨酸(Hyp),血清黄疸指数总胆汁酸(TBA)及总胆红素(TBIL)水平;测定各组大鼠胆汁流量并计算肝脏指数;采用HE染色检测肝组织病理变化。急性毒性试验研究中,40只KM小鼠随机分为对照组和黄肝煎液组,黄肝煎液组以最大给药剂量灌胃给予黄肝煎液,对照组灌胃给予等量生理盐水。观察给药后14 d内小鼠的急性毒性反应,并测定血清ALT、AST、ALP、肌酐(CRE)、尿素氮(BUN)、白蛋白(ALB)水平,计算脏器指数,评价黄肝煎液的急性毒性。结果:与模型组比较,黄肝煎液能够显著降低大鼠血清肝功能指标(ALT、AST、ALP和Hyp)及黄疸指数(TBIL和TBA),明显减小肝脏系数,增加胆汁流量,差异具有统计学意义(P<0.01),肝组织病理学改变显著改善。急性毒性试验中,以最大给药浓度1.63 g·mL^(-1),最大给药体积40 mL·kg^(-1)灌胃小鼠,24 h内给予黄肝煎液3次,最大日给药量为195.6 g·kg^(-1)(相当于临床成人日用剂量的80倍)时,除部分小鼠体质量降低、稀便软便外,其余小鼠均未出现死亡和明显的急性毒性反应。结论:黄肝煎液对ANIT诱导的大鼠胆汁淤积性肝损伤具有一定的保护作用,且无明显急性毒性反应。

Objective:To study the protective effects on cholestatic liver injury and the acute toxicity of Huanggan decoctions.Methods:A total of 72 SD rats were randomly divided into control group,model group,positive control group,Huanggan decoction low,medium and high dose groups,with 12 rats in each group.The rats were given corresponding drugs by continuous intragastric administration for 14 days,twice a day.On the 12th day,α-naphthalene isothiocyanate(ANIT)was administered to establish the cholestatic liver injury rat model,except for the control group.A fully automated biochemical analyzer was used to measure the serum liver function indexes(ALT,AST,ALP and Hyp)and the serum jaundice indexes(TBIL and TBA).The bile flow and the liver index were determined.Liver histopathological changes were observed by hematoxylin-eosin(HE)staining.The acute toxicity of Huanggan decoctions was investigated by a maximum dose method.A total of 40 KM mice were randomly divided into control group and Huanggan decoction group.Mice in the Huanggan decoction group were given Huanggan decoctions by intragastric administration with maximum doses,and mice in the control group were given the same amount of normal saline.Then the acute toxic reaction of mice within 14 days were observed and recorded,the serum levels of ALT,AST,ALP,CRE,BUN and ALB were determined,and organ indexes were calculated.Results:Compared with those in the model group,the serum levels of ALT,AST,ALP,TBIL,TBA and Hyp in the Huanggan decoction groups were decreased significantly,liver coefficients were significantly decreased,the bile flows were increased(P<0.01),and the liver histopathological changes were significantly improved.In the acute toxicity study,the mice were intragastrically administrated with Huanggan decoctions at the maximum concentration of 1.63 g·mL^(-1) and the maximum volume of 40 mL·kg^(-1),for 3 times in 24 h,the maximum daily dose was up to 195.6 g·kg^(-1)(equivalent to 80 times of the clinical daily dose for adults).No death or acute toxicity was observed except for some mice with reduced body mass and loose stools.Conclusion:Huanggan decoctions exerted a good hepatoprotective effect on ANIT-induced cholestasis liver injury in rats,and no obvious toxic and side effects were observed.