UGT1A10基因SNP rs10187694对吗替麦考酚酯代谢的影响

发布时间：2018-06-02 17:44

本文选题：尿苷二磷酸葡萄糖醛酸转移酶 + 霉酚酸酯　；参考：《中国现代医学杂志》2017年12期

【摘要】：目的体外研究尿苷二磷酸葡萄糖醛酸转移酶(UGT1A10)基因SNP rs10187694多态位点G/A变异对吗替麦考酚酯(MMF)代谢的影响,明确该位点变异是否与MMF个体代谢差异有关。方法采用基因重组、定点突变技术,构建UGT1A10基因SNP rs10187694位点含有不同等位基因的重组过表达载体,POLO 3000转染法将重组过表达载体转染入HEK293细胞,采用液相色谱-质谱联用仪(LC/MS/MS)系统,检测霉酚酸(MPA)代谢产物7-O-葡醛酸苷(MPAG)的生成量,以评估转染不同等位基因HEK293细胞中酶的活性。结果成功构建pIRES2-EGFP-prom(G)和pIRES2-EGFP-prom(A)重组过表达载体,并转染HEK293细胞。LC/MS/MS系统检测结果显示,携带A等位基因的突变型UGT1A10代谢MMF产生MPAG的能力降低,其24 h MPAG的生成量为(226.00±14.57)nmol/L,而野生型的生成量为(269.00±14.07)nmol/L,UGT1A10突变型MPAG的生成量是野生型的84.00%,两者比较,差异有统计学意义(P0.05)。结论 UGT1A10基因SNP rs10187694位点G/A突变可影响MMF代谢产物MPAG的生成,可能是造成不同个体MMF代谢差异的重要原因之一。
[Abstract]:Objective to study the effect of SNP rs10187694 polymorphism of uridine diphosphate glucuronyltransferase (UGT1A10) gene on the metabolism of mycophenolate mofetil (MMF) in vitro, and to determine whether this mutation is related to the individual metabolic difference of MMF. Methods using gene recombination and site-directed mutagenesis technique, a recombinant overexpression vector containing different alleles in SNP rs10187694 locus of UGT1A10 gene was constructed. The recombinant overexpression vector was transfected into HEK293 cells with different alleles. The recombinant overexpression vector was transfected into HEK293 cells by liquid chromatography-mass spectrometry (LC-MS) system. The production of 7-Ouronic acid glucuronide (7-Ouronoside) metabolite was determined to evaluate the enzyme activity in HEK293 cells transfected with different alleles. Results Recombinant expression vectors pIRES2-EGFP-prompromG and pIRES2-EGFP-prompromA) were successfully constructed and transfected into HEK293 cells. The results of MS / MS system analysis showed that the ability of mutant UGT1A10 carrying A allele to metabolize MMF to produce MPAG was decreased. The production of 24 h MPAG was 226.00 卤14.57 nmol / L, while that of wild type was 269.00 卤14.07 nmol / L, the production of UGT1A10 mutant MPAG was 84.00% of that of wild type, the difference was statistically significant (P 0.05). Conclusion the G / A mutation at SNP rs10187694 site of UGT1A10 gene may affect the production of MMF metabolite MPAG, which may be one of the important reasons for the difference of MMF metabolism in different individuals.
【作者单位】：上海交通大学附属第一人民医院普外科;
【基金】：国家自然科学基金(No:81270557,81110108010)
【分类号】：R96

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