RESUMEN
The present study was designed to determine the effects of Puer tea and green tea on blood glucose level. Male BALB/c mice were administered green tea extract (GTE) or Puer tea extract (PTE), either intragastrically or in their drinking water. The major components of these teas are epigallocatechin gallate (EGCG) and caffeine, respectively. Blood glucose measurement results showed that mice fed intragastrically or mice that drank GTE, PTE or caffeine showed significantly lower blood glucose levels compared to the control group. However, EGCG exhibited no influence on the blood glucose levels. When caffeine was eliminated from the GTE and PTE, the effect on the blood glucose levels was abolished, but the effect was recovered when caffeine was re-introduced into the extracts. Evaluation of hematological and biochemical indices at the time of the greatest caffeine-induced decrease in blood glucose levels showed that the effect of caffeine was specific. Microarray analyses were performed in 3T3-L1 preadipocytes and mature adipocytes treated with 0.1 mg · mL(-1) caffeine to identify factors that might be involved in the mechanisms underlying these effects. The results showed that few genes were changed after caffeine treatment in adipocytes, and of them only phospholipid transfer protein (PLTP) may be ralated to blood glucose. In conclusion, this study indicates that caffeine may be the key constituent of tea that decreases blood glucose levels, and it may be used to treat type 2 diabetes.
Asunto(s)
Animales , Masculino , Ratones , Células 3T3-L1 , Adipocitos , Metabolismo , Glucemia , Metabolismo , Cafeína , Farmacología , Camellia sinensis , Química , Hipoglucemiantes , Farmacología , Ratones Endogámicos BALB C , Proteínas de Transferencia de Fosfolípidos , Metabolismo , Extractos Vegetales , Farmacología , TéRESUMEN
AIM@#(-)-Epigallocatechin-3-gallate (EGCG), a major compound of tea polyphenols, exhibited antitumor activity in previous studies. In these studies, EGCG usually inhibits EGFR, and impairs the ERK1/2 phosphorylation in tumor cells. The aim was to clarify the mechanism of ERK1/2 activation induced by EGCG.@*METHOD@#Jurkat and 293T cells were treated with EGCG in different culture conditions. Western Blotting (WB) was employed to analyze ERK1/2 and MEK phosphorylation. Cetuximab and FR180204 were used to inhibit cell signaling. The stability of EGCG was assessed by HPLC. The concentration of hydrogen peroxide generated by the auto-oxidation of EGCG was determined by photocolorimetric analysis.@*RESULTS@#Activation of ERK1/2 was observed to be both time-and dose-dependent. Stimulation of cell signaling was dependent on MEK activity, but independent of EGFR activity. Unexpectedly, EGCG was depleted within one hour of incubation under traditional culture conditions. Auto-oxidation of EGCG generated a high level of hydrogen peroxide in the medium. Addition of catalase and SOD to the acidic medium inhibited the oxidation of EGCG. However, this particular condition also prevented the phosphorylation of ERK1/2. The generation of ROS by hydrogen peroxide may also induce ERK1/2 activation in Jurkat cells.@*CONCLUSION@#ERK1/2 phosphorylation was caused by auto-oxidation of EGCG. Traditional culture conditions were determined to be inappropriate for EGCG research.
Asunto(s)
Humanos , Camellia sinensis , Química , Catalasa , Metabolismo , Catequina , Farmacología , Peróxido de Hidrógeno , Metabolismo , Células Jurkat , Sistema de Señalización de MAP Quinasas , Proteína Quinasa 3 Activada por Mitógenos , Metabolismo , Proteína Quinasa 6 Activada por Mitógenos , Metabolismo , Oxidación-Reducción , Fosforilación , Extractos Vegetales , Farmacología , Polifenoles , Farmacología , Superóxido Dismutasa , MetabolismoRESUMEN
Human cytomegalovirus is an important pathogen which widely infects human. In this study, an eukaryotic expression vector containing human cytomegalovirus gB680 and pp65m genes was injected into BALB/c mice to explore the immune response. Firstly, the recombinant plasmid pVAX1/gB680+pp65m and plasmid pVAX1 were transfected into COS-7 cells respectively and the transiently expressed product was detected by RT-PCR and Western blot. The maxiprepared plasmid pVAX1/gB680 + pp65m and pVAX1 by alkaline lysis with SDS and purified by Sepharose 4FF column chromatography were then used to immunize BALB/c mice and the humoral and cellular immune responses were determined. Recombinant plasmid pVAX1/gB680+pp65m could be expressed in COS-7 cells, and could induce antibodies and cellular immune responses in BALB/c mice.