两点采样法评估人体NAT2酶的基因型与异烟肼代谢表型、肝功能异常之间的关系  

Two-point sampling method to evaluate the relationship between human NAT2 enzyme genotype and isoniazid metabolic phenotype and hepatitis

作　　者：刘国辉[1] 王艳梅 曾坚 曹炜鹏 张红梅[1] 王平[1] LIU Guo-hui;WANG Yan-mei;ZENG Jian;CAO Wei-peng;ZHANG Hong-mei;WANG Ping(The Third People's Hospital of Shenzhen,Shenzhen,Guangdong 518029,China;Shenzhen Hospital of University of Hong Kong,Shenzhen,Guangdong 518048,China)

机构地区：[1]深圳市第三人民医院肺病一科,广东深圳518029 [2]香港大学深圳医院内科,广东深圳518048

出　　处：《中国热带医学》2022年第8期718-723,共6页China Tropical Medicine

基　　金：广东省高水平临床重点专科(深圳市配套建设经费)项目(No.SZGSP010)。

摘　　要：目的通过两点采样法评估NAT2酶的基因型与异烟肼的乙酰化率、肝损害之间的关系。方法选取口服异烟肼达到稳态的患者共57例,分别于服药后2、4 h采取末梢血样。采用超高效液相色谱质谱联用设备(UPLC-MS)测定其中的异烟肼和乙酰异烟肼的药物浓度,计算二者的比率值。任意时间点低于1的比值定义为慢代谢临床表型,其余为中间代谢表型和快代谢临床表型。同时对血样标本提取核酸,测定的N-乙酰转移酶-2(NAT2)的rs1041983点位的SNP情况。根据SNP结果,将基因型分为慢代谢组(TT)、中间代谢组(CT)和快代谢组(CC)。将基因型与代谢表型、肝功能损害发生之间分别进行比对,判断NAT2的药物代谢表型与基因突变类型之间的关系。结果慢代谢与快代谢人群中,两个时间点的药物浓度均有高低,两者之间的可信区间有重叠,无法明确区分。单点代谢产物之间的比值不能全部区分出慢代谢人群。而两个时间点的代谢产物之间的比值可以完全区分快慢代谢人群的基因型。慢代谢基因型人群发生肝功能异常的发生率约84.6%,快代谢人群发生率仅29.5%,两者差异有统计学意义(χ^(2)=12.485,P<0.001)。结论NAT2慢代谢基因型人群发生肝损害的人群比例较高。但基因型只能提示代谢速率的快慢,而不能确定是否必定会发生肝功能损害。两点采样的乙酰异烟肼与异烟肼的比值可以直接作为判断NAT2酶快慢代谢的诊断指标,更具有实际应用价值。最佳参考的比率取值为1。ObjectiveTo evaluate the relationship between NAT2(N-acetyltransferase 2)enzyme genotype and isoniazid acetylation rate and ATDILI(antituberculosis drug-induced liver injury)by multi-point limited sampling method.MethodsA total of 57 patients who took isoniazid(INH)orally and reached a steady-state were selected.Peripheral blood samples were taken 2 h and 4 h after taking the drug.The concentrations of INH and Acetyl-INH were determined by UPLC-MS(Ultraperformance liquid chromatography-mass spectrometry)and the subsequent Acetyl-INH/INH metabolic ratio was calculated.The ratio at any time point below 1.0 was defined as slow acetylators,and the others were intermediate,fast acetylators.DNA of the subjects was extracted from blood samples,and the SNP(The single nucleotide polymorphisms)situation of rs1041983 of NAT2 was sequenced.The genotypes were divided into slow metabolizers(TT),intermediate metabolizers(CT)and fast metabolizers(CC)based on SNP results.The relationship between NAT2 genotype,metabolic phenotype,and the occurrence of ATDILI was analyzed.ResultsDuring the populations with slow acetylators and fast acetylators,the Acetyl-INH/INH metabolic ratios at both time points were high and low,and the confidence intervals between them overlapped and could not be exactly distinguished.The single-time ratios cannot distinguish slow metabolizers(TT),but the ratios at two-time points can exactly distinguish the genotypes of fast and slow metabolizers.The rates of ATDILT between slow metabolizers and fast metabolizers were 84.6%and 29.5%,with a significant difference(χ^(2)=12.485,P<0.001).ConclusionsThe proportion of people with NAT2 slow metabolizers with ATDILT is higher.However,the genotype can only indicate the speed of the metabolic rate,and cannot determine whether ATDILT must occur.The Acetyl-INH/INH metabolic ratio of multi-point sampling can be directly used as a diagnostic tool to detect the genotype of NAT2,which has a more practical application value.The reference ratio takes a value of 1.

关 键 词：结核 基因型 异烟肼 N-乙酰转移酶-2 肝炎 代谢表型 

分 类 号：R521[医药卫生—内科学]