RESUMO
This study provides the first example of a strategy to design a practical ligand toward lysosomal acid α-glucosidase (GAA) focusing on N-alkyl derivatives of 1,4-dideoxy-1,4-imino-d-arabinitol (DAB). The optimized N-4'-(p-trifluoromethylphenyl)butyl-DAB (5g) showed a Ki value of 0.73 µM, which was 353-fold higher affinity than N-butyl-DAB (3f) without a terminal phenyl group. Docking analysis showed that the phenyl part of 5g was accommodated in a lipophilic pocket. Furthermore, the p-trifluoromethyl group effectively suppresses the fluctuation of the phenyl group, allowing it to produce a stable bonding form with GAA. 5g increased the midpoint of the protein's protein denaturation temperature (Tm) by 6.6 °C above that in the absence of the ligand and acted as a "thermodynamic stabilizer" to improve the thermal stability of rhGAA. 5g dose-dependently increased intracellular GAA activities in Pompe patient's fibroblasts with the M519V mutation; its effect was comparable to that of DNJ, which is under clinical trials.
Assuntos
Doença de Depósito de Glicogênio Tipo II , alfa-Glucosidases , Humanos , alfa-Glucosidases/metabolismo , Doença de Depósito de Glicogênio Tipo II/tratamento farmacológico , Doença de Depósito de Glicogênio Tipo II/genética , Doença de Depósito de Glicogênio Tipo II/metabolismo , Ligantes , Lisossomos/metabolismo , FibroblastosRESUMO
This study aimed to investigate the effects of interleukin-25, which belongs to the interleukin-17 family, on short-term high-fructose diet-induced hepatic triacylglycerol accumulation. Rats were fed a high-starch (control) or high-fructose diet for 7 d, with or without intraperitoneal administration of recombinant interleukin-25 from days 3-7. Treatment with interleukin-25 significantly reduced the mRNA levels and activity of fatty acid synthesis enzymes and caused a nominal reduction in hepatic triacylglycerol levels in rats fed a high-fructose diet but not in those fed a control diet. Interleukin-25 treatment did not affect the mRNA levels of ß-oxidation enzymes in either the control or fructose-fed rats. These results suggest that treatment with interleukin-25 suppresses short-term high-fructose diet-induced fatty acid synthesis and leads to the alleviation of triacylglycerol accumulation in the liver.
Assuntos
Frutose , Interleucina-17 , Fígado , Animais , Ratos , Dieta , Ácidos Graxos/metabolismo , Frutose/farmacologia , Expressão Gênica , Interleucina-17/farmacologia , Fígado/efeitos dos fármacos , Fígado/metabolismo , Ratos Wistar , RNA Mensageiro/metabolismo , Triglicerídeos/metabolismoRESUMO
We investigated the effects of dietary supplementation with sodium butyrate (NaB) on the lipid levels, gene expression, and proteins related to lipid metabolism in nonalcoholic fatty liver disease (NAFLD) rat models fed a high-sucrose diet for 3 weeks. Supplementation with 1% and 3% NaB reduced high-sucrose-induced hepatic triacylglycerol levels and expression of genes and proteins related to fatty acid synthesis, such as fatty acid synthase and malic enzyme, in a dose-dependent manner. NaB supplementation did not affect hepatic cholesterol levels or expression of genes related to ß-oxidation. NaB may prevent high-sucrose-induced NAFLD by repressing the fatty acid synthesis pathway.
Assuntos
Hepatopatia Gordurosa não Alcoólica , Animais , Ácido Butírico/farmacologia , Dieta , Suplementos Nutricionais , Fígado/metabolismo , Hepatopatia Gordurosa não Alcoólica/tratamento farmacológico , Hepatopatia Gordurosa não Alcoólica/etiologia , Hepatopatia Gordurosa não Alcoólica/metabolismo , Ratos , Sacarose/efeitos adversos , Triglicerídeos/metabolismoRESUMO
Human promyelocytic HL60 cells can be differentiated into macrophagelike cells by treatment with 12Otetra decanoylphorbol13acetate (TPA). Certain 5' upstream regions of the zinc finger protein (ZNF)encoding genes contain duplicated GGAA motifs, which are frequently found in the TPAresponding gene promoter regions. To examine transcriptional responses to TPA, 5'flanking regions of human zinc finger CCCHtype containing, antiviral, ZNF252, ZNF343, ZNF555, ZNF782 and zinc finger nfx1type containing 1 (ZNFX1) genes were isolated by polymerase chain reaction (PCR) and ligated into a multiplecloning site of the pGL4.10[luc2] vector. Transient transfection and a luciferase assay revealed that the ZNFX1 promoter most prominently responded to the TPA treatment. Deletion and point mutation experiments indicated that the duplicated GGAA motif in the 100bp region positively responded to TPA. In addition, reverse transcriptionquantitative PCR and western blotting showed that the mRNA and protein of ZNFX1 accumulate during the differentiation of HL60 cells. These results indicated that expression of the TPAinducible ZNFX1 gene, which belongs to the group of interferonresponsive genes, is regulated by the cisaction of the duplicated GGAA motif.