Abstract
RAW 264.7 cells and HUVECs were compared to evaluate the effects of dialyzed coffee extract (DCE) and artificial coffee (AC). Immunoprecipitation high performance liquid chromatography (IP-HPLC) showed DCE-2.5- (equivalent to 2.5 cups of coffee a day) and DCE-5-induced protein expression that was beneficial to human health, i.e., they led to significant increases in proliferation-, immunity-, cellular protection-, antioxidant signaling-, and osteogenesis-related proteins but decreases in inflammation-, NFkB signaling-, cellular apoptosis-, and oncogenic signaling-related proteins in RAW 264.7 cells, and slight decreases in angiogenesis-related proteins in HUVECs. These protein expression changes were less frequently observed for DCE-10 treatment, while AC treatment induced very different changes in protein expression. We suggest that the favorable cellular effects of DCE were derived from minor coffee elements that were absent in AC, and that the reduced effects of DCE-10 compared with those of DCE-2.5 or DCE-5 might have been caused by greater adverse reactions to caffeine and chlorogenic acid in DCE-10 than DCE-2.5 or DCE-5. IP-HPLC results suggested that minor coffee elements in DCE might play beneficial roles in the global protein expression of proliferation-, immunity-, anti-inflammation-, cell protection-, antioxidant-, anti-apoptosis-, anti-oncogenesis-, and osteogenesis-related proteins in RAW 264.7 cells and enhance anti-angiogenic signaling in HUVECs.
Highlights
Coffee is a favorite drink worldwide, and many authors have investigated the effects of caffeine and chlorogenic acids in clinical and cell-based experiments
The expression of general growth factors, that is, transforming growth factor- β1 (TGF-β1), IGFIIR, and ERβ, changed the minimum to less than ±5% to the control housekeeping proteins (Fig. 3C2). These results suggested that the increased expression of GH, GHRH, and ERβ by dialyzed coffee extract (DCE)-2.5 or DCE-5 might be associated with enhanced RAS signaling and positively impact human health and that the marked reductions observed in the expression of HER1, HER2, and IGFIIR after DCE-2.5 or DCE-5 treatment might be useful for the treatment of different human diseases, such as breast cancer and diabetes
The expression of pAKT (88.7%), ERK-1 (92.2%), and phosphatidylinositol-3-kinases (PTEN, 89.4%) was slightly decreased after artificial coffee (AC) treatments (Fig. 3D2). These results suggested that in RAW 264.7 cells treated with DCE, nuclear factor kappa-light-chainenhancer of activated B cells (NFkB) signaling was not affected and the cells were in an unstressful condition to the non-treated control, whereas treatment with AC increased NFkB signaling, increased the expression of tumor necrosis factor-α (TNFα) and MDR, and decreased the expression of pAKT, ERK-1, and PTEN, indicating increased cellular stress
Summary
Coffee is a favorite drink worldwide, and many authors have investigated the effects of caffeine and chlorogenic acids (major components of coffee) in clinical and cell-based experiments. In protein expression in macrophages, which can engulf coffee elements in vitro. Preliminary studies on the bio-compatibility of DCE and RAW 264.7 cells showed no cytotoxic effect and little antigenic stimulation within a range of DCE doses[19,20]. Multiple trials have shown that IP-HPLC can detect protein expression changes accurately and reproducibly (±5% standard deviation). IP-HPLC was used to assess the expression of different functional proteins (n = 189) in RAW 264.7 cells treated with DCE or AC (1 mM chlorogenic acid and 2 mM caffeine) in vitro
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