c-Myc抑制剂10058-F4减轻脂多糖/右旋半乳糖胺诱导的小鼠急性肝损伤

c-Myc Inhibitor 10058-F4 Attenuates Lipopolysaccharide/D-galactosamine-induced Acute Liver Injury in Mice

本研究探讨c-Myc抑制剂10058-F4对脂多糖(lipopolysaccharide,LPS)和右旋半乳糖胺(D-galactosamine,D-Gal)诱导的急性肝损伤的影响及其可能机制。32只雄性BALB/c小鼠(Mus musculus)分为4组:正常对照组、10058-F4组、LPS/D-Gal组和10058-F4+LPS/D-Gal组。通过腹腔注射LPS/D-Gal诱导小鼠急性肝损伤,10058-F4在注射LPS/D-Gal前0.5 h经腹腔注入。注射LPS/D-Gal 1.5 h后处死小鼠并采集样本,分别采用实时荧光定量PCR法和ELISA法检测肝组织中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的mRNA水平和血清中TNF-α的水平。另取32只小鼠经上述相同处理后,注射LPS/D-Gal 6 h后处死并采集样本,检测小鼠血清中转氨酶活性及白介素-6(interleukin-6,IL-6)的水平、肝组织中IL-6的mRNA水平及肝组织中半胱氨酸天冬氨酸特异性蛋白酶(cysteinyl aspartate specific proteinase,Caspase),包括Caspase-3、Caspase-8和Caspase-9的活性,苏木精-伊红(hematoxylin-eosin,HE)染色观察肝组织病理学变化,TUNEL法检测肝细胞凋亡。结果显示,c-Myc抑制剂10058-F4可以显著下调LPS/D-Gal暴露小鼠血清中丙氨酸氨基转移酶(alanine aminotransferase,ALT)和天冬氨酸氨基转移酶(aspartate aminotransferase,AST)水平,降低血清和肝组织中的TNF-α与IL-6水平,抑制肝组织中Caspase-3、Caspase-8和Caspase-9的活性,减少TUNEL阳性细胞数以及肝组织病理学改变。以上结果提示,c-Myc抑制剂10058-F4可通过抑制炎症反应和减少肝细胞凋亡在LPS/D-Gal诱导的急性肝损伤中发挥保护作用。

This study is to investigate the effects of the c-Myc inhibitor 10058-F4 in acute liver injury induced by lipopolysaccharide(LPS)and D-galactosamine(D-Gal)and its underlying mechanism.32 male BALB/c mice(Mus musculus)were randomly divided into four groups:control group,10058-F4 group,LPS/D-Gal group and 10058-F4+LPS/D-Gal group.Acute liver injury was induced by intraperitoneal injection of LPS/D-Gal,and 10058-F4 was injected intraperitoneally 0.5 h prior LPS/D-Gal.The mice were sacrificed at 1.5 h post LPS/D-Gal exposure,and the mRNA level of tumor necrosis factor-α(TNF-α)in liver and the level of TNF-αin serum were determined by quantitative real-time PCR and ELISA,respectively.Another 32 mice were prepared and undergone the same above-mentioned procedure,then sacrificed at 6 h post LPS/D-Gal exposure,and the levels of transaminases and interleukin-6(IL-6)in serum were determined.The mRNA level of IL-6 and the activities of cysteinyl aspartate specific proteinase(Caspase),including Caspase-3,Caspase-8 and Caspase-9 in li-ver were determined.In addition,hematoxylin-eosin(HE)staining was used to observe the histopathological changes and hepatocellular apoptosis was observed by TUNEL assay.The results showed that treatment with the c-Myc inhibitor 10058-F4 in LPS/D-Gal-exposed mice significantly decreased the levels of serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST),decreased the mRNA levels of TNF-αand IL-6 in liver,and reduced the serum levels of TNF-αand IL-6.The 10058-F4-treatment also suppressed the acti-vation of Caspase-3,Caspase-8 and Caspase-9,reduced the count of TUNEL-positive hepatocytes and alleviated the degree of liver lesions.In conclusion,the c-Myc inhibitor 10058-F4 alleviated LPS/D-Gal-induced acute liver injury,these protective benefits might be associated with the compromised inflammation and suppressed apoptosis.