乳酸脱氢酶抑制剂减轻内毒素诱导的小鼠急性肝损伤

Lactate dehydrogenase inhibitor alleviates LPS/D-Gal-induced acute liver injury in mice

目的探讨乳酸脱氢酶抑制剂对内毒素诱导的小鼠急性肝损伤的影响。方法将BALB/c小鼠分为4组:溶剂对照组,乳酸脱氢酶抑制剂NHI-2对照组,脂多糖(LPS)/D-半乳糖胺(D-Gal)组,LPS/D-Gal+NHI-2组。LPS/D-Gal组及LPS/D-Gal+NHI-2组小鼠腹腔注射LPS(10μg·kg^(-1))及D-Gal(700 mg·kg^(-1))诱导急性肝损伤,NHI-2在LPS/D-Gal暴露前30 min经腹腔注射NHI-2(30 mg·kg^(-1))。LPS/D-Gal暴露1.5 h或6 h后处死取肝组织及血清,检测血清中乳酸、冬氨酸转氨酶(ALT)、血清丙氨酸转氨酶(AST)及肿瘤坏死因子α(TNF-α)的水平,检测肝组织中丙二醛(MDA)含量及caspase-3/8/9活性;HE染色评估肝组织损伤情况;TUNEL染色评估肝细胞凋亡情况;采用生存曲线记录实验小鼠存活率。结果NHI-2干预可明显降低模型小鼠血清乳酸水平;与LPS/D-Gal组相比,LPS/D-Gal+NHI-2组血清TNF-α浓度无明显差异,但血清ALT、AST水平明显下降、肝组织结构损伤明显减轻、肝组织MDA含量明显下调、肝内caspase-3/8/9活性明显降低、TUNEL阳性细胞数明显减少;NHI-2干预也可明显提升模型小鼠生存率。结论乳酸脱氢酶抑制剂可减轻内毒素诱导的小鼠急性肝损伤。

Aim To investigate the effect of lactate dehydrogenase inhibitor on LPS/D-Gal-induced acute liver injury in mice.Methods BALB/C mice were divided into four groups:solvent control group,lactate dehydrogenase inhibitor NHI-2 group,lipopolysaccharide(LPS)/D-galactosamine(D-Gal)group and LPS/D-Gal+NHI-2 group.To induce acute liver injury,mice were injected intraperitoneally with LPS(10μg·kg^(-1))and D-Gal(700 mg·kg^(-1)),NHI-2 was intraperitoneally injected 30 min before LPS/D-Gal exposure.Liver tissue and serum were harvested 1.5 or 6 h after LPS/D-Gal exposure,serum lactate,serum aspartate aminotransferase(ALT),serum alanine aminotransferase(AST),serum tumor necrosis factor alpha(TNF-α)liver malondialdehyde(MDA)and liver caspase-3/8/9 levels were determined.HE staining was used to evaluate the degree of liver injury.TUNEL staining was used to evaluate hepatocyte apoptosis.Survival curve was used to record survival situation of tested mice.Results Serum lactate level of model mice was significantly reduced after treatment with NHI-2.Compared with LPS/D-Gal group,level of serum TNF-αshowed no significant difference,but serum ALT and AST level of LPS/D-Gal+NHI-2 group significantly decreased,injury of liver structure was remarkably attenuated,level of MDA and activity of caspase-3/8/9 in liver were significantly down-regulated,and the number of TUNEL-positive cells was significantly reduced.Treatment with NHI-2 also significantly improved the survival rate of LPS/D-Gal-insulted mice.Conclusion Lactate dehydrogenase inhibitor alleviates LPS/D-Gal-induced acute liver injury in mice.