α-酮戊二酸减轻脂多糖/D-半乳糖胺诱导的小鼠急性肝损伤

Alpha-Ketoglutarate Attenuates Endotoxin/D-Galactosamine-Induced Acuteliver Injury in Mice

该文旨在探讨α-酮戊二酸对脂多糖(lipopolysaccharide,LPS)及D-半乳糖胺(d-galactosamine,D-Gal)诱导的急性肝损伤发生发展的影响及其可能机制。实验分组:正常对照组、AKG单独处理组、LPS/D-Gal组、LPS/D-Gal+AKG组。在雄性BALB/c小鼠中,经腹腔注射LPS/D-Gal诱导急性肝损伤,α-酮戊二酸在LPS/D-Gal注射前0.5 h经腹腔注入。6 h后,处死动物,采集动物血浆及肝脏,采用比色法检测血浆转氨酶水平以评估肝损伤情况;采用HE染色观察肝组织病理学改变;采用TUNEL染色和比色法检测caspase-3、caspase-8和caspase-9的活性;Western blot检测切割型caspase-3(cleaved caspase-3)片段含量,反映肝细胞凋亡情况。结果显示,α-酮戊二酸干预显著下调LPS/D-Gal暴露小鼠血浆中天冬氨酸氨基转氨酶(aspartate transaminase,AST)与丙氨酸氨基转氨酶(alanine transaminase,ALT)水平,有效降低肝组织中caspase-3、caspase-8和caspase-9的活性以及切割型caspase-3片段的含量,明显减轻TUNEL阳性肝细胞数目以及肝组织病理学改变。以上结果提示,α-酮戊二酸可减轻LPS/D-Gal诱导的肝细胞凋亡及肝组织损伤。

The paper aimed to explore the effect of alpha-ketoglutarate on the occurrence and development of acute liver injury induced by LPS(lipopolysaccharide)and D-Gal(D-galactosamine)and the underlying mechanism.Experimental groups:Control,AKG group,LPS/D-Gal group,LPS/D-Gal+AKG group.In male BALB/c mice,acute liver injury was induced by intraperitoneal injection of LPS/D-Gal.Alpha-ketoglutarate was injected intraperitoneally 0.5 h prior LPS/D-Gal.The mice were sacrificed at 6 h after LPS/D-Gal challenging,and plasma samples were collected for measuring the level of plasma transaminase by colorimetry to evaluate the liver injury.The pathological changes of liver tissue were observed by HE staining.The activities of caspase-3,caspase-8 and caspase-9 were detected by colorimetry.The apoptotic cell was estimated by TUNEL assay,and the level of cleaved caspase-3 was detected by Western blot to imply hepatocyte apoptosis.The results showed that alpha-ketoglutarate-treated LPS/D-Gal-exposed mice had significantly down-regulated incidence of histologic lesions,lower plasma AST(aspartate aminotransferase)and ALT(alanine aminotransferase).Treatment with alpha-ketoglutarate also reduced cleaved caspase-3 and caspase-3,caspase-8,caspase-9 activities and reduced the count of TUNEL-positive hepatocytes.Alpha-ketoglutarate can attenuate the pathological changes of acute liver injury induced by LPS/D-Gal and exert the protective effect of anti-apoptosis.