Table 1.
Primer sequence.
Fig 1.
BBR treatment improves OGTT in diabetic mice.
A. Plasma glucose concentrations were determined at 0, 30, 60 and 120 min after oral glucose administration of glucose as described in Materials and Methods. B. The area under the OGTT curve. Data are shown as the mean ± S.E.M (n = 12). **p<0.01 vs Control; #p<0.05 vs DM; $p<0.05 vs LB.
Table 2.
Biochemical parameters of diabetic mice treated with BBR.
Fig 2.
BBR treatment improves liver morphology, liver index and reduces TC and TG in diabetic mice.
Liver morphology (A), TC and TG (B) and liver index (C) was determined in Control, DM, LB and HB animals as described in Materials and Methods. Data represent the mean ± SEM (n = 12). *p<0.05 vs Control, #p<0.05 vs DM, $p<0.05 vs LB.
Fig 3.
BBR treatment attenuates the alteration in gluconeogenesis and lipid metabolism and expression of HNF-4α and miR122 in liver of DM animals.
Expression of G6Pase, PEPCK, SREBP-1, FAS-1, CPT-1, ACCα, pACCα, HNF-4α, β-actin and miR122 were determined in Control, DM, LB and HB animals as described in Materials and Methods. A. A representative blot showing G6Pase, PEPCK, SREBP-1, FAS-1, CPT-1, ACCα, pACCα, HNF-4α and β-actin. B. Relative protein expression of G6Pase and PEPCK. C. Relative protein expression of SREBP-1, FAS-1 and CPT-1. D. Relative protein expression of ACCα and pACCα. E. Relative protein expression of HNF-4α. F. Relative protein expression of serum and liver miR122. Data represent the mean ± SEM (n = 12). *p<0.05, **p<0.01 vs Control; # p<0.05 vs DM; $p<0.05 vs LB.
Fig 4.
BBR treatment attenuates the alteration in gluconeogenesis and lipid metabolism and expression of HNF-4α and miR122 in PA-treated HepG2 cells.
HepG2 cells were incubated in the absence or presence of 0.3 mM PA or 0.3 mM PA with 10 μM BBR for 24 h and expression of G6Pase, PEPCK, SREBP-1, FAS-1, CPT-1, ACCα, pACCα, HNF-4α, β-actin and miR122 were determined as described in Materials and Methods. A. A representative blot showing G6Pase, PEPCK, SREBP-1, FAS-1, CPT-1, ACCα, pACCα, HNF-4α and β-actin. B. mRNA expression of G6Pase and PEPCK. C. Relative protein expression of G6Pase and PEPCK. D. mRNA expression of SREBP-1, FAS-1 and CPT-1. E. Relative protein expression of SREBP-1, FAS-1 and CPT-1. F. Relative protein expression of ACCα and ACCα/pACCα. G. mRNA expression of ACCα. H. mRNA expression of HNF-4α. I. Protein expression of HNF-4α. J. miR-122 expression in HepG2 cells. Data represent the mean ± SEM (n = 12). *p<0.05, **p<0.01 vs Control; #p<0.05 vs PA-treated HepG2 cells.
Fig 5.
BBR regulates expression of hepatic gluconeogenesis and lipogenic enzymes through HNF4α.
HepG2 cells were incubated in the absence or presence of HNF-4α plasmid (HNF-4α(+)) or HNF-4α plasmid plus 10 μM BBR (BBR 10 μM) or HNF-4α plasmid plus miR122 inhibitor (miR122(-)) and the expression of gluconeogenic and lipid metabolism enzymes examined. A. A representative blot showing G6Pase, PEPCK, SREBP-1, FAS-1, CPT-1, ACCα, pACCα and β-actin. B. Protein expression of G6Pase and PEPCK. C. mRNA expression of G6Pase and PEPCK. D. protein expression of SREBP-1, FAS-1 and CPT-1. E. mRNA expression of SREBP-1, FAS-1 and CPT-1. F. protein expression of ACCα and ACCα/pACCα. G. mRNA expression of ACCα. Data represent the mean ± SEM (n = 5). *p<0.05 vs Control; #p<0.05 vs HNF4α(+).
Fig 6.
The BBR mediated of expression of hepatic gluconeogenesis and lipogenic genes is not regulated through miR122.
HepG2 cells were incubated in the absence or presence of HNF-4α siRNA (HNF-4α(-)) or HNF-4α siRNA plus 10 μM BBR (BBR 10 μM) or HNF-4α siRNA plus miR122 plasmid (miR122(+)) and the expression of gluconeogenic and lipid metabolism enzymes examined. A. A representative blot showing G6Pase, PEPCK, SREBP-1, FAS-1, CPT-1, ACCα, pACCα and β-actin. B. Protein expression of G6Pase and PEPCK. C. mRNA expression of G6Pase and PEPCK. D. protein expression of SREBP-1, FAS-1 and CPT-1. E. mRNA expression of SREBP-1, FAS-1 and CPT-1. F. protein expression of ACCα and ACCα/pACCα. G. mRNA expression of ACCα. Data represent the mean ± SEM (n = 5). *p<0.05 vs Control; #p<0.05 vs HNF4α(-).