一氧化碳释放分子-2对小鼠急性肝功能衰竭保护作用的实验研究

An experimental study in protective effect of CORM-2 on acute liver failure in mice

目的探讨一氧化碳释放分子-2对小鼠急性肝功能衰竭(ALF)的保护作用及机制。方法雄性C57BL/6小鼠30只随机分为对照组、模型组和保护组,每组10只。通过一次性腹腔注射脂多糖/D-氨基半乳糖(LPS/D-GalN)构建小鼠ALF的动物模型,CORM-2于造模前30 min行尾静脉注射,造模后6 h分别留取血清、肝组织标本。生化分析仪检测血清中ALT、AST水平,HE观察肝脏病理改变,酶联免疫吸附法(ELISA)检测血清中TNF-α和IL-6的水平,反转录酶-聚合酶链反应(RT-PCR)检测肝组织中TNF-α和IL-6 mRNA的表达。结果保护组小鼠血清ALT[(1274.60±157.24)U/L比(3499.00±136.19)U/L]和AST[(1151.50±244.58)U/L比(4079.50±481.11)U/L]水平均明显低于模型组(均t1=33.81,t2=17.16,P<0.05);与模型组比较,保护组肝组织炎性细胞浸润明显减少,肝细胞坏死程度明显减轻[(0.14±0.05)比(0.37±0.05),t=10.29,P<0.05];保护组小鼠血清和肝组织中TNF-α[(139.60±28.39)pg/mL比(447.34±128.17)pg/mL、(0.31±0.03)比(0.69±0.05)]和IL-6[(215.21±85.16)pg/mL比(1461.58±244.90)pg/mL、(0.33±0.03)比(0.72±0.05)]表达水平明显低于模型组(均t1=7.41,t2=20.61,t3=15.20,t4=21.15,P<0.05)。结论 CORM-2能够抑制小鼠ALF时的炎性反应,减轻肝脏病理损伤,其机制可能与抑制促炎细胞因子TNF-α和IL-6的释放有关。

Objective To investigate the protective effect and mechanisms of carbon monoxide releasing molecule（CORM-2）on acute liver failure（ALF）in mice.Methods Male C57BL/6 mice were randomly divided into three groups：control, model and protected group. The ALF animal models were induced by intraperitoneal injection of lipopolysaccharide/D-galactosamine（LPS/D-GalN）.CORM-2 was administered via tail vein30 minutes before LPS/D-GalN treatment.All serum samples and liver specimens were collected for detection in6 hours after the administration of LPS/DGalN.The serum levels of alanine aminotransferase（ALT）and aspartate aminotransferase（AST）were measured by automatic biochemical analyzer.The liver pathologic changes were observed by light microscopy using hematoxylin and eosin（HE）staining.The serum levels of tumor necrosis factor-α（TNF-α）and interleukin-6（IL-6）were measured by enzyme linked immunosorbent assay（ELISA）.The expression of TNF-αand IL-6 mRNA in the liver tissues were measured by reverse transcript polymerase chain reaction（RT-PCR）in each group.Results The serum levels of ALT [（1274.60±157.24）U/L vs（3499.00±136.19）U/L]and AST [（1151.50±244.58）U/L vs（4079.50± 481.11）U/L]in protected group were significantly lower than those in model group（t1=33.81,t2=17.16 both P0.05）.Massive inflammatory cells infiltration and hepatocyte necrosis were widely spread in model group,while liver injuries were obviously relieved in protected group[（0.14±0.05）vs（0.37±0.05）,t=10.29,P0.05].The serum levels of TNF-α [（139.60±28.39）pg/mL vs（447.34±128.17）pg/mL]and IL-6 [（215.21±85.16）pg/mL vs（1461.58±244.90）pg/mL]in protected group were significantly reduced compared with model group（t1=7.41,t2=20.61 both P0.05）.TNF-αmRNA [（0.31±0.03）vs（0.69±0.05）]and IL-6 mRNA [（0.33±0.03）vs（0.72±0.05）]measured by RT-PCR in live tissues of protected group were also significantly lower than that in model group（both P0.05）.Conclusion CORM-2 could remarkably inhibit the inflammatory activity and attenuate liver injury,mechanism of which might be related to inhibiting the release of proinflammatory cytokines TNF-αand IL-6.