目的:探讨miR-376b-3p对HepG2细胞Ⅱ相代谢酶UGT2B7的调控作用。方法:构建含有UGT2B7序列的重组质粒并进行双荧光素酶基因活性检测;利用荧光定量PCR技术检测miR-376b-3p对UGT2B7 mRNA表达的影响;使用蛋白印迹法检测miR-376b-3p对UGT2B7...目的:探讨miR-376b-3p对HepG2细胞Ⅱ相代谢酶UGT2B7的调控作用。方法:构建含有UGT2B7序列的重组质粒并进行双荧光素酶基因活性检测;利用荧光定量PCR技术检测miR-376b-3p对UGT2B7 mRNA表达的影响;使用蛋白印迹法检测miR-376b-3p对UGT2B7蛋白表达的影响。结果:与miR-376b-3p阴性对照组相比,在HEK293T细胞中共转染miR-376b-3p与UGT2B7 3′-UTR野生型质粒组荧光素酶活性降低13%,差异具有统计学意义(P<0.05),共转染miR-376b-3p与UGT2B7 3′-UTR突变体质粒组的荧光素酶活性差异无统计学意义(P>0.05);与阴性对照组相比,在HepG2细胞中转染miR-376b-3p模拟物组的UGT2B7 m RNA水平下降44%,转染miR-376b-3p抑制剂组升高41%,差异均具有统计学意义(P<0.05);与阴性对照组相比,在HepG2细胞中转染miR-376b-3p模拟物组的UGT2B7蛋白表达下降34%,转染miR-376b-3p抑制剂组升高67%,差异均具有统计学意义(P<0.05)。结论:miR-376b-3p与UGT2B7 3′-UTR直接结合,通过降解UGT2B7 m RNA、抑制UGT2B7蛋白翻译而进行转录后调控。展开更多
AIM: To clarify whether Mycophenolic acid(MPA) or its metabolite AcMPAG can cause gastrointestinal disturbances, and to explore the effect of UGT2B7 SNP 211G>T on the pharmacokinetic of AcMPAG. METHODS: Twenty-four...AIM: To clarify whether Mycophenolic acid(MPA) or its metabolite AcMPAG can cause gastrointestinal disturbances, and to explore the effect of UGT2B7 SNP 211G>T on the pharmacokinetic of AcMPAG. METHODS: Twenty-four renal transplant patients were enrolled in this study. Pharmacokinetic study was performed on day 14 after transplantation and symptoms were recorded on the same day. Multiple blood samples were collected before dosing and 0.5,1,1.5,2,4,6,8,10 and 12 hours after morning dosing. Plasma concentrations of MPA, MPAG and AcMPAG were detected by HPLC. Genotype of UGT2B7 211G>T was determined using PCR-RFLP method. RESULTS: No significant difference was observed between patients with and without side effects for AUC(0-12) of MPA and MPAG. The values of AUC(0-12)/dose of MPA, MPAG and AcMPAG were (39.7±12.3), (1063.7±646.5) and (10.2±3.0) ng·h·mL-1·mg-1 in TT genotype group and (42.0±20.0),(1466.5±683.3) and (20.7±5.6) ng·h·mL-1·mg-1 in GG genotype group, respectively. The ratio of AUCAcMPAG(0-12)/AUCCMPA(0-12), which was used to describe the change of UGT2B7 enzyme activity, was 0.49±0.20,0.34±0.26 and 0.28±0.14 in recipients with 211GG, GT and TT genotype, respectively(P<0.05). CONCLUSION: There was no significant differences in pharmacokinetics of MPA and its metabolites between side effect group and non-side effect group. It seems that the polymorphism 211G>T in the UGT2B7 gene results in the reduction of enzyme acitivity.展开更多
文摘目的:探讨miR-376b-3p对HepG2细胞Ⅱ相代谢酶UGT2B7的调控作用。方法:构建含有UGT2B7序列的重组质粒并进行双荧光素酶基因活性检测;利用荧光定量PCR技术检测miR-376b-3p对UGT2B7 mRNA表达的影响;使用蛋白印迹法检测miR-376b-3p对UGT2B7蛋白表达的影响。结果:与miR-376b-3p阴性对照组相比,在HEK293T细胞中共转染miR-376b-3p与UGT2B7 3′-UTR野生型质粒组荧光素酶活性降低13%,差异具有统计学意义(P<0.05),共转染miR-376b-3p与UGT2B7 3′-UTR突变体质粒组的荧光素酶活性差异无统计学意义(P>0.05);与阴性对照组相比,在HepG2细胞中转染miR-376b-3p模拟物组的UGT2B7 m RNA水平下降44%,转染miR-376b-3p抑制剂组升高41%,差异均具有统计学意义(P<0.05);与阴性对照组相比,在HepG2细胞中转染miR-376b-3p模拟物组的UGT2B7蛋白表达下降34%,转染miR-376b-3p抑制剂组升高67%,差异均具有统计学意义(P<0.05)。结论:miR-376b-3p与UGT2B7 3′-UTR直接结合,通过降解UGT2B7 m RNA、抑制UGT2B7蛋白翻译而进行转录后调控。
文摘AIM: To clarify whether Mycophenolic acid(MPA) or its metabolite AcMPAG can cause gastrointestinal disturbances, and to explore the effect of UGT2B7 SNP 211G>T on the pharmacokinetic of AcMPAG. METHODS: Twenty-four renal transplant patients were enrolled in this study. Pharmacokinetic study was performed on day 14 after transplantation and symptoms were recorded on the same day. Multiple blood samples were collected before dosing and 0.5,1,1.5,2,4,6,8,10 and 12 hours after morning dosing. Plasma concentrations of MPA, MPAG and AcMPAG were detected by HPLC. Genotype of UGT2B7 211G>T was determined using PCR-RFLP method. RESULTS: No significant difference was observed between patients with and without side effects for AUC(0-12) of MPA and MPAG. The values of AUC(0-12)/dose of MPA, MPAG and AcMPAG were (39.7±12.3), (1063.7±646.5) and (10.2±3.0) ng·h·mL-1·mg-1 in TT genotype group and (42.0±20.0),(1466.5±683.3) and (20.7±5.6) ng·h·mL-1·mg-1 in GG genotype group, respectively. The ratio of AUCAcMPAG(0-12)/AUCCMPA(0-12), which was used to describe the change of UGT2B7 enzyme activity, was 0.49±0.20,0.34±0.26 and 0.28±0.14 in recipients with 211GG, GT and TT genotype, respectively(P<0.05). CONCLUSION: There was no significant differences in pharmacokinetics of MPA and its metabolites between side effect group and non-side effect group. It seems that the polymorphism 211G>T in the UGT2B7 gene results in the reduction of enzyme acitivity.
基金supported by grants from the Major State Basic Research Development Program of China("973"Program)(2011CB510000)the National Natural Science Foundation of China(81130021,81171198,81200870,81000542)
