异甘草酸镁对刀豆蛋白A诱导的急性肝衰竭小鼠模型的影响

Effect of magnesium isoglycyrrhizinate on concanavalin A-induced acute liver failure in mice

目的研究异甘草酸镁(Mg IG)对刀豆蛋白A(Con A)诱导的急性肝衰竭小鼠模型的作用及机制。方法选取健康BALB/C小鼠90只,随机分为4组,分别为对照组、Mg IG对照组,每组各10只;Con A模型组、Mg IG+Con A预处理组,每组各35只(其中15只用来计算总生存率)。提取小鼠肝脏及外周血,测定生存率、血清转氨酶水平,肝组织HE染色评价小鼠肝损伤程度,TUNEL荧光染色及caspase-3蛋白活性检测评价小鼠肝细胞凋亡水平,液相芯片法检测小鼠血清炎症细胞因子IL-1β和TNFα含量。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 Con A模型组小鼠总生存率为40%,Mg IG+Con A预处理组小鼠总生存率为80%。Con A模型组血清ALT与AST水平较对照组均显著上升(P值均<0. 01),Mg IG+Con A预处理组血清ALT与AST水平较Con A模型组均显著降低(P值均<0. 01)。对照组、Mg IG对照组、Con A模型组及Mg IG+Con A预处理组病理评分分别为1. 0±0. 2、1. 2±0. 3、3. 7±0. 6、2. 3±0. 5,组间比较差异有统计学意义(F=2. 7,P <0. 05);凋亡细胞/100细胞数值分别为0. 2±0. 1、0. 1±0. 1、7. 8±1. 3、2. 2±0. 4,组间比较差异有统计学意义(F=27. 6,P <0. 001);caspase-3活性分别为0. 813±0. 022、0. 930±0. 033、1. 347±0. 042、1. 060±0. 053,组间比较差异有统计学意义(F=51. 072,P <0. 001)。Con A模型组与对照组比较,血清IL-1β与TNFα水平均显著上升(P值均<0. 05)。Mg IG+Con A预处理组与Con A模型组比较,血清IL-1β与TNFα均显著下降(P值均<0. 05)。结论异甘草酸镁对刀豆蛋白A诱导的小鼠急性肝衰竭具有明显保护作用,减少肝细胞凋亡并减轻IL-1β和TNFα引起的肝脏炎症反应可能是其缓解肝损伤的机制。

Objective To investigate the effect of magnesium isoglycyrrhizinate(MgIG)on concanavalin A(ConA)-induced acute liver failure in mice and its mechanism. Methods A total of 60 healthy BALB/C mice were randomly divided into control group with 10 mice,MgIG control group with 10 mice,ConA model group with 20 mice,and MgIG-ConA pretreatment group with 20 mice.The liver and peripheral blood were collected to evaluate survival rate,serum levels of aminotransferases,and degree of liver injury by HE staining of liver tissue;TUNEL fluorescence staining and activity of caspase-3 protein were used to evaluate the apoptosis of hepatocytes;the liquid chip method was used to measure the serum levels of the inflammatory cytokines interleukin-1β(IL-1β)and tumor necrosis factorα(TNFα).A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups. Results The overall survival rate of mice was 40%in the ConA model group and 80%in the MgIG-ConA pretreatment group.The ConA model group had significantly increased serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)compared with the control group(both P<0.01),while the MgIG+ConA pretreatment group had significantly reduced serum levels of ALT and AST compared with the ConA model group(both P<0.01).The control group,the MgIG control group,the ConA model group,and the MgIG-ConA pretreatment group had a pathological score of 1.0±0.2,1.2±0.3,3.7±0.6,and 2.3±0.5,respectively,and there was a significant difference between groups(F=2.7,P<0.05);the apoptotic cells/100 cells ratio was 0.2±0.1,0.1±0.1,7.8±1.3,and 2.2±0.4,respectively,in these four groups,and there was a significant difference between groups(F=27.6,P<0.001);the activity of caspase-3 protein was 0.813±0.022,0.930±0.033,1.347±0.042,and 1.060±0.053,respectively,in these four groups,and there was a significant difference between groups(F=51.072,P<0.001).Compared with the control group,the ConA model group had significant increases in serum levels of IL-1βand TNFα(both P<0.05);compared with the ConA model group,the MgIG+ConA pretreatment group had significant reductions in serum levels of IL-1βand TNFα(both P<0.05). Conclusion MgIG exerts a marked protective effect against ConA-induced acute liver failure in mice and can alleviate liver injury possibly by reducing hepatocyte apoptosis and alleviating liver inflammatory response caused by IL-1βand TNFα.