黄芩苷对急性药物性肝损伤小鼠的保护机制

Effect of baicalin against acetaminophen-induced acute liver injury in mice and its possible mechanism

目的:探讨黄芩苷对急性药物性肝损伤小鼠的保护机制。方法:2019年6月至2019年12月,以陕西省人民医院实验中心提供的6~8周C57BL/6雄性小鼠进行实验。采用腹腔注射对乙酰氨基酚(APAP)诱导小鼠急性药物性肝损伤模型。按随机数字表法分为空白对照组(Control组)、药物肝模型组(APAP组)及黄芩苷治疗组(BA+APAP组),处理16 h后取血标本及肝脏组织,检测各组血清谷丙转氨酶(ALT)活性、天门冬氨酸氨基转移酶(AST)活性;肝组织丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性;血清肿瘤坏死因子-α(TNF-α)水平、白细胞介素-6(IL-6)水平;观察肝脏病理形态学改变;检测肝组织中细胞外信号调节激酶(ERK)及磷酸化ERK(p-ERK)的表达变化。组间比较采用单因素方差分析。结果:光镜下黄芩苷治疗组肝损伤程度较模型组明显减轻;黄芩苷治疗组血清ALT、血清AST明显低于模型组[(614.2±132.9)、(498.7±52.8)U/L比(6826.4±684.2)、(5860.7±464.8)U/L,t=44.862、67.850,P均<0.05];黄芩苷治疗组血清TNF-α、血清IL-6明显低于模型组[(156.8±44.3)、(673.1±43.7)pg/ml比(334.3±36.8)、(899.8±98.9)pg/ml,t=18.702、3.644,P均<0.05];芩苷治疗组肝匀浆MDA含量明显低于模型组[(6.3±1.6)nmol/mg蛋白比(9.7±1.6)nmol/mg蛋白,t=38.524,P<0.05];黄芩苷治疗组肝匀浆SOD活性明显高于模型组[(12.7±1.7)U/mg蛋白比(8.0±0.6)U/mg蛋白,t=4.854,P<0.05];黄芩苷治疗组肝组织p-ERK/ERK蛋白比值明显低于模型组(0.4±0.2比1.0±0.1,t=18.974,P<0.05)。结论:黄芩苷对APAP诱导的急性药物性肝损伤小鼠具有保护作用,其作用机制可能与减少ERK磷酸化有关。

Objective To investigate the protective effect of baicalin on acetaminophen(APAP)-induced acute liver injury in mice and the possible mechanisms.Methods From June 2019 to December 2019,15 male C57BL/6 mice were supplied by Animal Feeding Center of Shaanxi Provincial People′s Hospital.Mouse models of liver injury were established by intraperitoneal injection of APAP.Mice were randomly divided into three groups:blank control group,liver injury model group(APAP group),and baicalin+APAP group(BA+APAP group).Blood samples and liver tissues were collected at 16 h after APAP modeling to determine the serum aspartate transaminase(AST),alanine transaminase(ALT);malondialdehyde(MDA),superoxide dismutase(SOD);tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)levels.The pathological changes were observed.The expression of extracellular signal-regulated MAP kinases(ERK)and phosphorylation of ERK(p-ERK)in liver tissues was detected by Western blotting.One-way ANOVA was used for statistical analysis.Results The histopathological abnormalities were attenuated in BA+APAP group.The levels of ALT and AST in BA+APAP group were significantly lower than those in APAP group[(614.2±132.9),(498.7±52.8)U/L vs.(6826.4±684.2),(5860.7±464.8)U/L,t=44.862,67.850,both P<0.05].The levels of TNF-αand IL-6 in BA+APAP group were significantly lower than in APAP group[(156.8±44.3),(673.1±43.7)pg/ml vs.(334.3±36.8),(899.8±98.9)pg/ml,t=18.702,3.644,both P<0.05].The MDA level in BA+APAP group was significantly lower than that in APAP group[(6.3±1.6)nmol/mg·prot vs.(9.7±1.6)nmol/mg·prot,t=38.524,P<0.05].The SOD level in BA+APAP group was significantly lower than that in APAP group[(12.7±1.7)U/mg·prot vs.(8.0±0.6)U/mg·prot,t=4.854,P<0.05].The p-ERK/ERK level in BA+APAP group was significantly lower than that in APAP group(0.4±0.2 vs.1.0±0.1,t=18.974,P<0.05).Conclusion This study suggests that baicalin can protect APAP-induced acute liver injury,which may be related to down-regulation of expression of p-ERK.