核苷类似物拉米夫定与恩替卡韦对大鼠慢加急性肝衰竭的影响

Effects of lamivudine and entecavir on acute-on-chronic liver failure of rats

目的观察核苷类似物拉米夫定与恩替卡韦对大鼠实验性慢加急性肝衰竭(ACLF)的影响,并初步探讨其机制。方法将40只SD大鼠随机分为对照组8只、实验组32只。实验组腹腔注射40%CCl4及猪血清4周以诱导慢性肝损伤,将其再随机分为模型组8只、拉米夫定组12只、恩替卡韦组12只,均给予腹腔联合注射D-氨基半乳糖、脂多糖以诱导ACLF;诱导6 h后,拉米夫定组及恩替卡韦组分别给予拉米夫定10 mg/kg及恩替卡韦0.05mg/kg灌胃,每日1次,连续3 d。对照组给予相同体积生理盐水腹腔注射或灌胃,模型组大鼠予以相同体积生理盐水灌胃。分别于慢性肝损伤造模4周和诱导ACLF后24、72 h采血,采用全自动生化分析仪检测肝功能(ALT、AST、TBIL),ELISA法检测诱导ACLF后72 h血清肿瘤坏死因子α(TNF-α)和白细胞介素17(IL-17);诱导ACLF后72 h处死大鼠,肝组织切片行HE染色及Masson染色。结果慢性肝损伤造模4周,实验组肝功能指标均高于对照组(P均<0.05)。诱导ACLF后24 h,拉米夫定组和恩替卡韦组肝功能指标均高于模型组(P均<0.05),拉米夫定组肝功能指标均高于恩替卡韦组但差异无统计学意义。在诱导ACLF后72 h,模型组、拉米夫定组、恩替卡韦组肝功能指标较24 h时均下降(P均<0.01);但拉米夫定组血清ALT、TBIL均仍高于模型组(P均<0.05),拉米夫定组血清AST虽高于模型组但差异无统计学意义;恩替卡韦组肝功能指标虽均高于模型组但差异均无统计学意义,其肝功能指标虽均低于拉米夫定组但差异无统计学意义。诱导ACLF后72 h,模型组、拉米夫定组、恩替卡韦组血清TNF-α、IL-17均高于对照组,但此两种因子水平在四组间的差异均无统计学意义。拉米夫定、恩替卡韦组肝组织病理镜下所见与模型组大致相仿,但肝细胞局灶性坏死较模型组多见,可见片状坏死,且拉米夫定组较恩替卡韦组重。结论在ACLF急性发作期,应用核苷类似物拉米夫定和恩替卡韦均可能一过性加重肝组织的损伤,且以拉米夫定为重。

Objective To observe the effects of nucleoside analogueses lamivudine（ LAM） and entecavir（ ETV） on the acute-on-chronic liver failure（ ACLF） of rats and to investigate its mechanism preliminarily. Methods Forty SD rats were randomly divided into two groups： the control group（ 8 rats） and experimental group（ 32 rats）. The rats of the experimental group were treated with 40% CCl41 m L/kg ip biw and porcine serum 0. 5 m L ip qw for 4 weeks in order to induce the chronic liver injury. Then,the rats of the experimental group were randomly divided into the model group（ 8 rats）,LAM group（ 12 rats）,and ETV（ 12 rats）,and all rats of the three groups were given D-Gal ＋ LPS ip to induce ACLF. Six hours later,the rats of the LAM group and ETV group were treated with LAM 10 mg/kg ig qd or ETV 0. 05 mg/kg ig qd,respectively; the rats of the control group and model group were treated with the same volume of normal saline for 3 days. The liver function indexes including ALT,AST,and TBil were detected with the automatic biochemistry analyzer at 4 weeks after chronic liver injury and at 24 and 72 h after induction of ACLF. The slices of hepatic tissues were respectively dyed with the methods of HE and Masson staining,and the histopathological changes were checked with light microscope. The serum levels of TNF-α and IL-17 were detected by ELISA at 72 h after induction of ACLF. Results At 4 weeks after modeling of chronic liver injury,the serum levels of liver function indexes of the rats in the experimental group were higher than those of the control group（ all P〈0. 05）. At 24 h after induction of ACLF,the serum levels of liver function indexes of the LAM group and ETV group were all higher than those of the model group（ all P〈0. 01）; the levels of liver function indexes of the LAM group were all higher than those of the ETV group,but the differences between the two groups were not significant. At 72 h after induction of ACLF,the serum levels of three liver function indexes decreased in each experimental group（ all P〈0. 01）; the levels of ALT and TBil of the LAM group were still higher than those of the model group（ both P〈0. 05）,but the statistical difference in the AST level between the two groups was not significant; the levels of liver function indexes of the ETV group were higher than those of the model group,but no statistical differences were found between these two groups;what＇s more,there were no statistical differences in the levels of liver function indexes between the LAM group and ETV group. At 72 h after induction of ACLF,the serum levels of TNF-α and IL-17 of the model group,LAM group,and ETV groups were all higher than those of the control group,but there were no statistical differences in the levels of the two cytokines between these four groups. The hepatic histopathological changes of the LAM group and ETV group,observed in light microscope,were similar to those of the model group,but the focal necrosis of hepatic cells was more common than that of model group,and there were few of the piece necrosis; the hepatic histopathological changes of the LAM group were more serious than those of the ETV group. Conclusion In the period of acute attack of ACLF,the treatment of LAM or ETV may transiently aggravate the injury of hepatic tissues,and the effect of LAM is severer than that of ETV.