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目的探讨嘌呤类生物碱1,3,7,9-四甲基尿酸(Theacrine)对Sirtuin3(SIRT3)调节的脂肪酸代谢相关基因的影响。方法对3T3-L1前脂肪细胞进行诱导分化,并在诱导的第5天对细胞培养液中加入不同浓度的Theacrine孵育,采用实时荧光PCR(RT-PCR)方法测定细胞内SIRT3及脂肪酸代谢相关基因的表达。并对正常小鼠灌胃给予低、中、高剂量的Theacrine(10,20,40 mg·kg-1)及Caffeine(20 mg·kg-1)1h后,采用RT-PCR测定小鼠肝脏组织、棕色脂肪组织中SIRT3及脂肪酸代谢相关基因LCAD、ATP5c、UCP1、HTGL、FABP4等基因的表达,同时采用Western Blotting检测肝脏中SIRT3的蛋白表达。结果与正常对照组比较,Theacrine可以上调3T3-L1前脂肪细胞在分化过程中SIRT3、LCAD、FABP4、LPL的表达。低剂量的Theacrine可显著上调小鼠肝脏中SIRT3基因,高剂量则表现为抑制作用,与其蛋白表达结果一致。同时,低、高剂量的Theacrine分别显著提升及抑制小鼠肝脏中LCAD、FABP4、HTGL、ATP5c等基因表达。此外,低剂量的Theacrine对小鼠棕色脂肪中SIRT3、LCAD等基因表达的影响不大,但显著上调了UCP1的基因表达。结论一定剂量的Theacrine能调节相关基因影响脂肪酸代谢,该作用可能与其上调SIRT3的表达相关。
Objective To investigate the effects of purine alkaloids 1,3,7,9-tetramethyluric acid (SACT3) on fatty acid metabolism-related genes regulated by Sirtuin3 (SIRT3). Methods The 3T3-L1 preadipocytes were induced to differentiate and incubated with different concentrations of Theacrine on the 5th day after induction. Real-time fluorescence PCR (RT-PCR) was used to determine the intracellular SIRT3 and fatty acid metabolism related genes expression. The normal mice were intragastrically given low, medium and high doses of Theacrine (10,20,40 mg · kg-1) and Caffeine (20 mg · kg-1) for 1 h, and the liver tissues SIRT3, LCAD, ATP5c, UCP1, HTGL and FABP4 in brown adipose tissue were detected by Western Blotting. The protein expression of SIRT3 in liver was detected by Western Blotting. Results Compared with the normal control group, Theacrine up-regulated the expression of SIRT3, LCAD, FABP4 and LPL in the differentiation of 3T3-L1 preadipocytes. Low doses of Theacrine can significantly upregulate the SIRT3 gene in mice liver, high doses showed inhibition, consistent with its protein expression results. At the same time, low and high doses of Theacrine significantly increased and inhibited LCAD, FABP4, HTGL, ATP5c and other gene expression in mouse liver respectively. In addition, low dose of Theacrine on mouse brown fat SIRT3, LCAD and other gene expression has little effect, but significantly up-regulated UCP1 gene expression. Conclusions Theacrine dose-dependently regulates fatty acid metabolism, which may be related to its upregulation of SIRT3 expression.