RESUMEN
Herbal raw materials with antidiabetic activity can be a valuable support to therapy. An optimized extraction process allows for the best possible health-promoting effect. Box-Behnken design was employed to optimize the content of methanol used in the extraction mixture, its time, and temperature. The aim of this study was to enhance the efficiency of the pomegranate flowers extraction process in order to obtain extracts with the highest enzyme inhibition power (α-amylase and α-glucosidase), which is important for the antidiabetic effect and the highest antioxidant activity (DPPH assay). In the Box-Behnken design model, the content of pelargonidin-3,5-glucoside-anthocyanin compound that is associated with antidiabetic activity was also optimized as a variable associated with the action profile of pomegranate flower extracts. The process optimization carried out in this study provides a basis for further research using the pomegranate flower extract with the most potent desired properties, essential for supporting diabetes treatment based on pomegranate flowers.
Asunto(s)
Antocianinas , Granada (Fruta) , Antocianinas/análisis , Antocianinas/farmacología , Antioxidantes/química , Flores/química , Glucósidos/análisis , Hipoglucemiantes/análisis , Hipoglucemiantes/farmacología , Hipoglucemiantes/uso terapéutico , Metanol/análisis , Extractos Vegetales/química , alfa-Amilasas , alfa-GlucosidasasRESUMEN
This study was designed to optimize the extraction rate of total polyphenols and ellagic acid from pomegranate flowers. Single factors were investigated for liquid-to-material ratio (5-25), ethanol concentration (20%-60%), sonication time (5-60 min), and sonication power (150-500 W). The level range of the Box-Bokhen design was determined with respect to the single-factor results. The components of each index were normalized using the entropy weighting method for obtaining the comprehensive evaluation value. Under the actual conditions, the final optimization results were 17 for liquid-to-material ratio, 43% for ethanol concentration, 10 min for ultrasonic time, and 300 W for ultrasonic power. The extracts obtained under optimal conditions were tested for the inhibition of Streptococcus mutans and its biofilm, and results showed that pomegranate flowers exerted some inhibitory effects on the bacterium. Phosphomolybdenum and FRAP assays were used, and DPPH, ABTS, and O2- radical scavenging tests were conducted, indicating that pomegranate flower extracts have good antioxidant capacity.
Asunto(s)
Granada (Fruta) , Antioxidantes/farmacología , Ácido Elágico , Etanol , Flores , Extractos Vegetales , PolifenolesRESUMEN
OBJECTIVES: Pomegranate flower is a kind of uygur medicine with anti - type 2 diabetes, anti - lipid, anti - inflammation, anti - oxidation. We investigated the effect of pomegranate flower extract (PFE) on the proliferation, differentiation and apoptosis of 3T3-L1 preadipocytes, as well as the effects of five compounds in PF on cell differentiation. METHODS: 3T3-L1 preadipocytes were treated with PFE (0.5, 1, 2, 5, 10, 20, 50, 100 µg/mL), quercetin, luteolin, ursolic acid, apigenin and kaempferol (5, 10, 20, 40, 80 µM), and cell viability was measured at 24, 48 and 72 h by Cell Counting Kit-8. The modified cocktail induction method induced the differentiation of 3T3-L1 preadipocytes, and treated them with PFE and the compounds. The lipid accumulation was determined by oil red O staining, and the intracellular triglyceride content was determined by commercial kit. The expressions of PPARγ, C/EBP, LPL, DGAT and aP2 mRNA in mature adipocyte were determined by q-PCR, and the expressions of PPARγ, Akt, p-akt and PI3K protein were determined by western blot. 3T3-L1 preadipocytes were treated with PFE (5, 10, 20 µg/mL) while induced apoptosis by palmitate (300 µM), Hoechst staining to observe apoptosis morphology, Annexin â ¤- FITC/PI staining with flow cytometry instrument to detect the number of early and late apoptosis cells, the q-PCR and western blot for determining the Bcl-2, Bax, caspase 3 mRNA and protein expression. RESULTS: PFE (5, 10, 20 µg/mL) promoted or did not affect the proliferation and differentiation of 3T3-L1 preadipocytes, and reduced the number of early and late apoptotic cells, increased the expression of Bcl-2 mRNA and protein, and inhibited the expression of Bax and caspase-3 mRNA and protein. Furthermore, PFE (40, 60 µg/mL), quercetin (10, 20, 40 µM), luteolin (5, 10, 20 µM), apigenin(20,40 µM), kaempferol (20, 40 µM) significantly restrain the 3T3-L1 different extent proliferation and differentiation of preadipocyte, reduce the accumulation of lipids in adipocyte, reduce expression of adipogenesis factor, PFE(40, 60 µg/mL) inhibited the activation of the PI3K-Akt pathway by inhibiting the expression of PI3K and p-Akt proteins, and inhibited preadipocyte differentiation by reduce the expression of PPARγ protein. CONCLUSION: PFE has a concentration-related bidirectional effect on the proliferation, differentiation and apoptosis of 3T3-L1 preadipocytes, which depends on the regulation of PI3K-Akt pathway, which is of guiding role for PFE in the treatment of type 2 diabetes, hyperlipidemia, obesity and other diseases.