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目的:探讨微小RNA-373(microRNA-373,miR-373)通过调控SULT1A3的表达对芹菜素磺酸化代谢反应速率的影响。方法:通过在人结肠腺癌细胞Caco-2 TC7中转染miR-373 mimic构建miR-373高表达细胞模型;应用qRT-PCR方法检测miR-373高表达后细胞中SULT1A3 mRNA的表达水平;建立细胞裂解液与芹菜素反应体系,应用HPLC法测定各转染组中不同浓度(2.5,10和30μmol·L-1)芹菜素的代谢速率,并结合LC-MS/MS鉴定芹菜素及其代谢产物结构。结果:在miR-373高表达的Caco-2 TC7细胞中,SULT1A3的mRNA表达受到抑制;与阴性转染组(mimic negative control组,mimic NC组)相比,miR-373转染组(miR-373 mimic组)中不同浓度芹菜素的代谢速率下降(P<0.05);LC-MS/MS显示芹菜素代谢产物为单磺酸化结合产物。结论:miR-373通过靶向抑制SULT1A3,降低芹菜素的磺酸化代谢反应速率。
AIM: To investigate the effect of microRNA-373 (miR-373) on the rate of apigenin sulfonation by regulating the expression of SULT1A3. Methods: miR-373 mimic cells were transfected into human colon adenocarcinoma cell line Caco-2 TC7 to construct miR-373 overexpression cell model. The expression of SULT1A3 mRNA was detected by qRT-PCR after miR-373 overexpression was established. Cell lysate and apigenin reaction system, the metabolic rate of apigenin at different concentrations (2.5,10 and 30μmol·L-1) in each transfection group was determined by HPLC, and the combination of apigenin and its metabolites by LC-MS / MS Product structure. Results: The mRNA expression of SULT1A3 in Caco-2 TC7 cells overexpressing miR-373 was inhibited. Compared with the mimic negative control group (mimic NC group), miR-373 transfection group (miR- 373 mimic group) (P <0.05). The metabolites of apigenin were monosulfonated by LC-MS / MS. CONCLUSIONS: miR-373 decreases the rate of sulfation metabolism of apigenin by targeting SULT1A3.