Experimental Study Of Polymyxin B Combined With N-acetylcysteine In The Treatment Of Liver Injury Induced By α-amanita Toxophenin

Objective: To evaluate the efficacy of polymyxin B(POLB)combined with n-acetylcysteine(NAC)in the treatment of liver injury caused by alpha-amanotoxin(α-AMA).Objective: To evaluate the efficacy of polymyxin B(POLB)combined with n-acetylcysteine(NAC)in the treatment of liver injury caused by alpha-amanotoxin(α-AMA).Methods: Animal experimental research was carried out in two studies: short-term study(24h)and long-term study(14 days).Short-term study: Forty male mice in Kunming were randomly divided into 8 groups(n=5),which were normal saline(NS)group,POLB group,NAC group,POLB+NAC group,α-AMA group,α-AMA+POLB group,α-AMA+NAC group and α-AMA+POLB+NAC group.Intraperitoneal injection was used to treat mice with α-AMA toxin at 0.35mg/kg,and polymyxin B(2.5mg/kg)and n-acetylcysteine(200mg/kg)were given at 4h,8h and 12 h after exposure.24 h later,the eyeballs of mice were collected and blood was collected for biochemical detection(ALT,AST,BUN,Cr).After dissection,liver and kidney were collected for organ coefficient calculation and liver histopathology analysis.Long-term study: Grouping,poisoning and treatment plan were the same as short-term study.The general performance,survival,weight changes and food intake changes of mice in each group at14 days were observed and recorded.Results: Short-term experimental study: 1.Biochemical indexes(ALT,AST,Cr,BUN):ALT(99.4±10.46)U/L in α-AMA group and(99.6±22.01)U/L in NS group were similar with no statistical difference(P > 0.05).ALT(127.8±23.96)U/L inα-AMA+POLB group.α-AMA+NAC group(292.2±171.15)U/L,αama +POLB+NAC group(134.8±19.76)U/L,ALT of the three groups were higher than those of the infected group,but there was no statistical difference(P > 0.05),AST of NS group(191.8±39.97)U/L,The AST of α-AMA group was(196.6±23.02)U/L,and there was no significant difference between the two groups.The AST of α-AMA+NAC group was(373.6±89.39)U/L,and that of αama +POLB+NAC group was(285.4±34.29)U/L.The AST of α-AMA+POLB group was 298.4±80.39 U/L,and the AST of the three treatment groups was higher than that of the infected group,but there was no statistically significant difference.In terms of BUN,the BUN of NS group was(9.94±0.67)U/L,and that of α-AMA group was(7.48±0.19)U/L,and the BUN ofα-AMA+NAC group was(8.26±0.89)U/L.BUN(8.26±0.62)U/L inα-AMA+POLB+NAC group and(9.44±1.79)U/L in α-AMA+POLB group.BUN in the three treatment groups was higher than that in the infected group,but in pairs There was no statistical significance.In terms of Cr value,the Cr value of NS group was(11.8±0.66)U/L,and the Cr value of the infected group was(10.6±0.51)U/L.The Cr value of the former group was higher than that of the latter group,and the Cr value of the α-AMA+NAC group was(12±1.52)U/L.Cr(10.6±0.25)U/L in theα-AMA+POLB+NAC group and Cr(10.6±0.75)U/L in the α-AMA+POLB group.BUN levels of the three treatment groups were higher than those of the poisoned group,but there was no statistical significance between the two groups.2.Histopathological results of mouse liver and kidney: 24 h after exposure,there was no inflammatory infiltration,bleeding or necrosis in the liver of NS group,POLB group,NAC group,POLB+NAC group,α-AMA group,α-AMA+NAC group,α-AMA+POLB+NAC group.Only the liver tissue of α-AMA+POLB group showed focal aggregation of inflammatory cells.But no bleeding,necrosis;No inflammatory infiltration,bleeding or necrosis were observed in the kidney tissue of each infected group.3.Organ coefficient:Liver coefficient: The liver coefficient of NS group(0.067±0.004)was higher than that of α-AMA group(0.065±0.004),but there was no statistical difference.The liver coefficient of α-AMA group(0.065±0.001)was higher than that of α-AMA+POLB group(0.054±0.004),α-AMA+NAC group(0.059±0.006),αama +POLB+NAC group(0.058±0.005).The liver coefficient of α-AMA group(0.065±0.004)was statistically significant compared with α-AMA+POLB(0.054±0.004),but there was no statistically significant difference between α-AMA+NAC(0.059±0.006)and αama +POLB+NAC(0.058±0.005).There was no significant difference between α-AMA group andα-AMA+NAC and α-AMA+POLB+NAC group.Renal coefficient: The liver coefficient of NS group(0.009±0.0003)was lower than that of α-AMA group(0.011±0.0002),and the comparison was statistically significant.The α-AMA+POLB group(0.012±0.004)and α-AMA+NAC group(0.012±0.001)were larger than α-AMA group(0.011±0.0002),and the comparison was not statistically significant.There was no statistical significance in α-AMA+POLB+NAC group(0.011±0.001)compared with α-AMA group(0.011±0.0002).Long-term experimental studies: 1.General situation of mice:All the mice in the non-poisoned group showed no poisoning and died.The dead mice in the poisoned group showed poor appetite,isolation,hair,poor reaction,uncoordinated movement,epileptic tic tic,groan,coma and other symptoms 2 days after exposure.The surviving mice in the poisoned group also showed poor reaction,uncoordinated movement,and involuntary head rotation to varying degrees until the14 th day.2.Changes in food intake and body weight of mice: From 1 to 4 days after exposure,the average food intake of mice in NS group showed an increasing trend,while the average food intake of mice in poison and drug intervention groups showed an overall decreasing trend.From 4 to 14 days,the average food intake of mice in poison and drug intervention groups was basically the same,and the average food intake of mice in normal saline group was higher than that in poison and drug treatment groups,and the changes of body weight and food intake of mice were basically similar.4.Survival of animals: 4 mice died in α-AMA group and 3 mice died in α-AMA+POLB group;Four mice died in α-AMA+NAC group.4 mice died in α-AMA+POLB+NAC group,and 0 mice died in NS group,POLB group,NAC group and POLB+NAC group.Conclusion: Polymyxin B combined with n-acetylcysteine has no protective effect on liver injury induced by alpha-amanotoxin.