Oda Hiroaki, Okuda Yuji, Yoshida Yukiko, Kimura Noriko, Kakinuma Atsushi
Laboratory of Nutritional Biochemistry, Department of Applied Molecular Biosciences, Nagoya University, Nagoya 464-8601, Japan.
Laboratory of Nutritional Biochemistry, Department of Applied Molecular Biosciences, Nagoya University, Nagoya 464-8601, Japan.
Biochem Biophys Res Commun. 2015 Oct 23;466(3):306-11. doi: 10.1016/j.bbrc.2015.09.010. Epub 2015 Sep 5.
The regulatory mechanism of phosphoenolpyruvate carboykinase (GTP) (EC 4.1.1.32) (PEPCK) gene expression and gluconeogenesis by phenobarbital (PB), which is known to induce drug-metabolizing enzymes, was investigated. Higher level of PEPCK mRNA was observed in spherical rat primary hepatocytes on EHS-gel than monolayer hepatocytes on TIC (type I collagen). We found that PB directly suppressed PEPCK gene expression in spherical hepatocytes on EHS-gel, but not in those on TIC. PB strongly suppressed cAMP-dependent induction of PEPCK gene expression. Tyrosine aminotransferase (TAT), another gluconeogenic enzyme, was induced by cAMP, but not suppressed by PB. Chronic administration of PB reduced hepatic PEPCK mRNA in streptozotocin-induced diabetic and nondiabetic rats, and PB reduced blood glucose level in diabetic rats. Increased TAT mRNA in diabetic rats was not suppressed by PB. These results indicated that PB-dependent reduction is specific to PEPCK. From pyrvate challenge test, PB suppressed the increased gluconeogenesis in diabetic rats. PEPCK gene promoter activity was suppressed by PB in HepG2 cells. In conclusion, we found that spherical hepatocytes cultured on EHS-gel are capable to respond to PB to suppress PEPCK gene expression. Moreover, our results indicate that hypoglycemic action of PB result from transcriptional repression of PEPCK gene and subsequent suppression of gluconeogenesis.
已知苯巴比妥(PB)可诱导药物代谢酶,本研究探讨了其对磷酸烯醇丙酮酸羧激酶(GTP)(EC 4.1.1.32)(PEPCK)基因表达及糖异生的调节机制。在EHS凝胶上培养的球形大鼠原代肝细胞中观察到的PEPCK mRNA水平高于在TIC(I型胶原)上培养的单层肝细胞。我们发现PB直接抑制EHS凝胶上球形肝细胞中PEPCK基因的表达,但对TIC上的肝细胞无此作用。PB强烈抑制cAMP依赖性的PEPCK基因表达诱导。另一种糖异生酶酪氨酸转氨酶(TAT)可被cAMP诱导,但不受PB抑制。长期给予PB可降低链脲佐菌素诱导的糖尿病大鼠和非糖尿病大鼠肝脏中的PEPCK mRNA水平,且PB可降低糖尿病大鼠的血糖水平。糖尿病大鼠中升高的TAT mRNA不受PB抑制。这些结果表明,PB依赖性降低对PEPCK具有特异性。通过丙酮酸激发试验,PB抑制了糖尿病大鼠中增加的糖异生。在HepG2细胞中,PB抑制了PEPCK基因启动子活性。总之,我们发现EHS凝胶上培养的球形肝细胞能够对PB作出反应以抑制PEPCK基因表达。此外,我们的结果表明,PB的降血糖作用源于PEPCK基因的转录抑制及随后糖异生的抑制。