正交设计探讨一线抗结核药物致小鼠肝损伤相互作用研究

Orthogonal Design to Investigate the Interaction of First-line Anti-tuberculosis Drugs on Liver Injury in Mice

目的:抗结核药物所致肝损伤机理不明,成为临床治疗结核病的主要障碍。方法:采用四因子2水平L_(16)(2^(15))正交设计,灌胃利福平(RIF)0.3 g/kg、异烟肼(INH)0.15 g/kg、吡嗪酰胺(PZA)0.63 g/kg以及乙胺丁醇(EMB)0.38 g/kg单一成分或组合物,1次/d,连续1 w后取血清测定相关生化指标,同时计算其肝脏指数。结果:利福平0.3 g/kg在升高小鼠血清ALT、AST、TBil、DBil、TBA含量或活力、HDL/VLDL比值、肝指数和降低血清TG、TC、HDL-C、LDL-C、VLDL含量等指标发挥着主要作用;异烟肼0.15 g/kg则在降低小鼠血清ALT、AST、TBA、TG、HDL-C、LDL-C活力或含量,升高TBil、DBil含量以及肝指数等指标中发挥着重要作用;吡嗪酰胺0.63 g/kg则在调节小鼠血脂含量以及肝指数中发挥着重要作用;乙胺丁醇0.38 g/kg则仅在调节小鼠血清HDL含量中发挥重要作用。利福平0.3 g/kg和异烟肼0.15 g/kg联用时,两者在多个指标上均表现出明显的交互作用,包括肝功能、血脂水平。利福平0.3 g/kg和吡嗪酰胺0.63 g/kg联用时,主要在血清脂质表现出明显的交互作用。异烟肼0.15 g/kg和吡嗪酰胺0.63 g/kg联用时在调节小鼠血清脂质及肝脏指数等指标上发生明显的交互作用。异烟肼0.15 g/kg与乙胺丁醇0.38 g/kg联用则在调节小鼠血清DBil水平上发挥交互作用;利福平0.3 g/kg与乙胺丁醇0.38 g/kg联用,吡嗪酰胺0.63 g/kg与乙胺丁醇0.38 g/kg联用则无明显交互作用;利福平0.3 g/kg+异烟肼0.15 g/kg+吡嗪酰胺0.63 g/kg三者联用主要在调节小鼠血清脂质以及肝脏指数表现出明显的交互作用;利福平0.3 g/kg+异烟肼0.15 g/kg+乙胺丁醇0.38 g/kg三者联用主要在调节小鼠肝功能以及血脂部分指标上表现出明显的交互作用;异烟肼0.15 g/kg+吡嗪酰胺0.63 g/kg+乙胺丁醇0.38 g/kg则在调节小鼠血脂部分指标上表现出明显的交互作用,而利福平0.3 g/kg+吡嗪酰胺0.63 g/kg+乙胺丁醇0.38 g/kg联用以及四药联用均无明显的交互作用。结论:利福平0.3 g/kg、异烟肼0.15 g/kg、吡嗪酰胺0.63 g/kg以及乙胺丁醇0.38 g/kg之间组合使用,可显著改善部分药物所致肝功能及血脂异常指标,其内在机制还有待进一步阐明。在临床选择监测肝损伤指标时应注意排除干扰因素,以免漏诊。

Objective:The mechanism of liver injury induced by anti-tuberculosis drugs is unclear,which becomes the main barrier of clinical treatment on tuberculosis.Methods:Orthogonal Design of 4 factors and 2 levels L_(16)(2^(15))was used.Single component or different combinations of 0.3 g/kg of rifampicin(RIF),0.15 g/kg of isoniazid(INH),0.63 g/kg of pyrazinamide(PZA),and 0.38 g/kg of ethambutol(EMB)were administrated to mice once per day.After administration for 1 w,the serum biochemical indicators and liver indexes of mice were determined.Results:0.3 g/kg of RIF played a major role in increasing serum activities or contents of alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBil),direct bilirubin(DBil),and total bile acids(TBA),the ratio of high density lipoprotein/very low density lipoprotein(HDL/VLDL),and liver indexes,and reducing serum contents of triglycerides(TG),total cholesterol(TC),HDL-cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),and VLDL.0.15 g/kg of INH played an important role in reducing serum activities or contents of ALT,AST,TBA,TG,HDL-C and LDL-C,and increasing the contents of TBil,DBil and liver indexes.0.63 g/kg of PZA regulated blood lipid levels and liver indexes in mice.0.38 g/kg of EMB only showed effect on the serum content of HDL.When 0.3 g/kg of RIF was combined with 0.15 g/kg of INH,significant interactions were observed in multiple indicators,including liver function and blood lipid level.When 0.3 g/kg of RIF was combined with 0.63 g/kg of PZA,significant interaction was observed in serum lipid.When 0.15 g/kg of INH was combined with 0.63 g/kg of PZA,the significant interaction was observed in regulating serum lipid and liver indexes in mice.When 0.15 g/kg of INH was combined with 0.38 g/kg of EMB,the interaction was observed in regulating the serum level of DBiL in mice.When 0.3 g/kg of RIF was combined with 0.38 g/kg of EMB,or 0.63 g/kg of PZA was combined with 0.38 g/kg of EMB,neither interaction was observed.When 0.3 g/kg of RIF,0.15 g/kg of INH,and 0.63 g/kg of PZA were combined,interactions were observed in regulating serum lipid and liver indexes in mice.When 0.3 g/kg of RIF,0.15 g/kg of INH,and 0.38 g/kg of EMB were combined,interactions were observed on regulating liver function and part indexes of blood lipid in mice.When 0.15 g/kg of INH,0.63 g/kg of PZA,and 0.38 g/kg of EMB were combined,the interaction was observed on regulating parts indexes of blood lipid in mice.Nevertheless,when 0.3 g/kg of RIF,0.63 g/kg of PZA and 0.38 g/kg of EMB were combined,or four drugs were used in combination,no significant interaction was observed.Conclusion:The single component or different combinations of RIF(0.3 g/kg),NIH(0.15 g/kg),PZA(0.63 g/kg),and EMB(0.38 g/kg)significantly treated the abnormity of liver function and liver indexes induced by some anti-tuberculosis drugs.The internal mechanism remained to be further clarified.The interference factors should be eliminated to avoid missed diagnosis in clinical selection of liver injury indicators.