The Study Of Interleukin-37 Anti-Acetaminophen-Induced Liver Injury On Mice

Objective:To investigate the anti-inflammatory mechanism of interleukin-37 by establishing the animal model of acetaminophen induced acute liver injury on mice.Methods:The male mice were randomly allocated into four groups(n=16),normal control group(control group),the model group of acetaminophen-induced liver injury(model group),IL-37 intervention group(prevention group) and IL-37 group.Each group of mice was randomly allocated into two subgroups,and then the two subgroups were executed by the time point 12 h and 24 h.For the control group the normal saline was injected into the enterocoelia,and then the PBS solution was injected after two hours.For the Model group,the normal saline was injected in the enterocoelia,and then the acetaminophen solution(250mg/kg) was injected after two hours.For the prevention group,the IL-37 b solution(1ug) was injected in the enterocoelia,and then the acetaminophen solution(250mg/kg) was injected after two hours.For the IL-37 group,the IL-37 b solution(1ug) was injected in the enterocoelia,and then the PBS solution was injected after two hours.At the time point 12 h and 24 h,each subgroup of mice were executed by cervical dislocation after obtaining blood respectively,the liver was obtained after execution.The blood and the liver were used for the following tests:(1) testing the level of serum ALT and AST with Reitman Frankel assa;(2) testing the level of serum IL-1β,IL-6 and TNF-α by ELISA;(3) weighing liver tissue with electronic balance for the calculation of liver index;(4) observing the morphologic changes of liver tissue by HE stain;(5) testing the expression of TNF-α and i NOS protein in liver tissue by immuno histochemistry.Results:(1)The impact of the ALT level and AST level:the ALT levels of model group were significantly higher than those of control group at the time point 12 h and 24h(P<0.01);the AST levels of model group at time point 24 h were significantly higher than the control group(P<0.01);the ALT and AST levels of model group at time point 24 h were higher than 12h(P<0.05);the ALT levels of prevention group at time point 12 h and 24 h were significantly lower than the model group(P<0.01);the AST levels of model group at time point 24 h were significantly lower than the model group(P<0.01);the ALT and AST levels between IL-37 group and control group at time point 12 h and 24 h had no significantly statistics differences(P>0.05);(2)The impact of the levels of serum IL-1β,IL-6 and TNF-α:the levels of serum IL-1β,IL-6 and TNF-α of model group at time point 12 h and 24 h were higher than the control group(P<0.05);the levels of serum IL-1β and IL-6 of model group at time point 24 h were significantly higher than 12h(P<0.01);the levels of serum IL-1β,IL-6 and TNF-α of prevention group at time point 12 h and 24 h were higher than the control group(P<0.05);the levels of serum IL-1β,IL-6 and TNF-α of IL-37 group and control group at time point 12 h and 24 h had no statistics differences(P>0.05);the levels of serum IL-1β, IL-6 and TNF-α of IL-37 group at time point 12 h and 24 h had no statistics differences(P>0.05);the levels of serum IL-1β,IL-6 and TNF-α of prevention group at time point 12 h and 24 h were significantly lower than the model group(P<0.01);(3)The comparison of liver index:the liver index of model group at time point 12 h and 24 h were significantly lower than the control group(P<0.01);the liver index of model group at time point 12 h were higher 24h(P<0.05);the liver index of IL-37 group and control group at time point 12 h and 24 h had no statistics differences(P>0.05);the liver index of prevention group at time point 12 h and 24 h were significantly lower than the model group(P<0.01);(4)The comparison of the level of TNF-α and i NOS:the levels of TNF-α and i NOS of model group at time point 12 h and 24 h were significantly higher than the control group(P<0.01);the levels of TNF-α and i NOS of prevention group at time point 12 h and 24 h were higher than the control group(P<0.05);the levels of TNF-α and i NOS of IL-37 group and the control group at time point 12 h and 24 h had no statistics differences(P>0.05);the levels of TNF-α and i NOS of prevention group at time point 12 h and 24 h were significantly lower than the model group(P<0.01);the levels of i NOS of model group at time point 24 h were higher than 12h(P<0.05);the levels of TNF-α of model group at time point 24 h were lower than 12h(P<0.05).Conclusions:(1)Successfully established the animal model of acetaminophen induced acute liver injury on mice,in addition,proved that IL-37 can alleviate hepatic function damage on mice;(2) IL-37 can lower the levels of ALT,AST,IL-1β,IL-6 and TNF-α in the model group of acetaminophen-induced liver injury;(3) IL-37 can lower the live index, lower levels of TNF-α and i NOS in the model group of acetaminophen-induced liver injury, alleviate hepatic function damage;(4) IL-37 is harmless to liver.