Effect Of Delphinidin Research Articles

The dietary anthocyanin delphinidin prevents bone resorption by inhibiting Rankl-induced differentiation of osteoclasts in a medaka (Oryzias latipes) model of osteoporosis.

The anthocyanin delphinidin is a natural compound found as water-soluble pigment in coloured fruits and berries. Anthocyanin-rich diets have been proposed to have bone protective effects in humans and mice, but the underlying mechanisms remain unclear. In this study, we used a medaka (Oryzias latipes) osteoporosis model to test the effects of delphinidin on bone cells in vivo. In this model, inducible transgenic expression of receptor-activator of NF-kβ ligand (Rankl) leads to ectopic formation of osteoclasts and excessive bone resorption, similar to the situation in human osteoporosis patients. Using live imaging in medaka bone reporter lines, we show that delphinidin significantly reduces the number of osteoclasts after Rankl induction and protects bone integrity in a dose-dependent manner. Our in vivo findings suggest that delphinidin primarily affects the de novo differentiation of macrophages into osteoclasts rather than the recruitment of macrophages to sites of bone resorption. For already existing osteoclasts, delphinidin treatment affected their morphology, leading to fewer protrusions and a more spherical shape. Apoptosis rates were not increased by delphinidin, suggesting that osteoclast numbers were reduced primarily by impaired differentiation from macrophage progenitors and reduced maintenance of pre-existing osteoclasts. Importantly, and in contrast to previously reported cell culture experiments, no effect of delphinidin on osteoblast differentiation and distribution was observed in medaka in vivo. Our study is the first report on the effects of delphinidin on bone cells in fish embryos, which are a unique model system for compound testing that is suitable for live imaging of bone cell behaviour in vivo.

Dietary delphinidin inhibits human colorectal cancer metastasis associating with upregulation of miR-204-3p and suppression of the integrin/FAK axis

Delphinidin is a flavonoid belonging to dietary anthocyanidin family that has been reported to possess diverse anti-tumoral activities. However, the effects of delphinidin on colorectal cancer (CRC) cells and the underlying mechanisms are not fully understood. Thus, we aimed to investigate the anti-cancer activity of delphinidin in CRC cells and the underlying molecular mechanisms. The effects of delphinidin on the viability, metastatic characteristics, signaling, and microRNA (miR) profile of human CRC cell lines used were analyzed. In vivo metastasis was also evaluated using xenograft animal models. Our findings showed that delphinidin (<100 μM) inhibited the colony formation of DLD-1, SW480, and SW620 cells, but non-significantly affected cell viability. Delphinidin also suppressed the migratory ability and invasiveness of the tested CRC cell lines, downregulated integrin αV/β3 expression, inhibited focal adhesion kinase (FAK)/Src/paxillin signaling, and interfered with cytoskeletal construction. Analysis of the miR expression profile revealed a number of miRs, particularly miR-204-3p, that were significantly upregulated and downregulated by delphinidin. Abolishing the expression of one upregulated miR, miR-204-3p, with an antagomir restored delphinidin-mediated inhibition of cell migration and invasiveness in DLD-1 cells as well as the αV/β3-integrin/FAK/Src axis. Delphinidin also inhibited the lung metastasis of DLD-1 cells in the xenograft animal model. Collectively, these results indicate that the migration and invasion of CRC cells are inhibited by delphinidin, and the mechanism may involve the upregulation of miR-204-3p and consequent suppression of the αV/β3-integrin/FAK axis. These findings suggest that delphinidin exerts anti-metastatic effects in CRC cells by inhibiting integrin/FAK signaling and indicate that miR-204-3p may play an important role in CRC metastasis.

Delphinidin induces cytotoxicity and potentiates cytocidal effect in combination with arsenite in an acute promyelocytic leukemia NB4 cell line.

The effects of delphinidin were investigated by focusing on growth inhibition, cell cycle arrest and apoptosis induction in the human acute promyelocytic leukemia (APL) NB4 cell line. Delphinidin exhibited a dose- and time-dependent cytotoxic effect against NB4 cells. Almost no cell cycle arrest, but an apparent increase in the percentage of sub-G1 cells was observed in delphinidin-treated cells. The activation of caspase-8 and -9 was observed as early as 1-h post-exposure to delphinidin, followed by the activation of caspase-3 from 3-h post-exposure. A substantial decrease in the expression level of Bid was also observed as early as 1-h post-exposure. A modest decrease in the mitochondrial membrane potential (ΔΨm) was observed at 3-h post-exposure, followed by a substantial time-dependent decrease in ΔΨm in treated cells. Delphinidin exerted more potent cytotoxicity against NB4 cells than normal peripheral blood mononuclear cells (PBMNCs). In addition, delphinidin in combination with an arsenic derivative arsenite (As(III)), which has demonstrated marked efficacy in patients with APL, achieved an enhanced cytocidal effect against NB4 cells, but lesser on PBMNCs. Treatment of NB4 cells with As(III) plus delphinidin did not increase, but decreased slightly, intracellular arsenic accumulation (As[i]) as compared to that treated with As(III) alone. These results suggested that delphinidin selectively sensitized NB4 cells to As(III), resulting in the enhancement of As(III) cytotoxicity by strengthening intrinsic/extrinsic pathway-mediated apoptosis induction, rather than affecting the As[i] levels. These observations may offer a rationale for the use of delphinidin to improve the clinical efficacy of As(III).

0397 : Delphinidin and calcium signaling in human T lymphocytes: a basis to elucidate pathways leading to proliferation and apoptosis

Delphinidin, an anthocyanin present in red wine, has been reported to exert vasculoprotective properties due to its vasodilatory and antioxidant effects at the level of the endothelium. However, the effect of delphinidin on cells involves in inflammation in cardiovascular diseases remains unknown. We have investigated the effect of delphinidin on the proliferation and apoptosis of T lymphocytes with respect to calcium signaling. Human T cells were isolated from blood of healthy patients. Proliferation was assessed using CyQUANT® NF Cell Proliferation Assay Kit and apoptosis by flow cytometry. Calcium signaling was analyzed using Fluo-4 probe and the mechanisms were depicted using appropriate drugs. Delphinidin did not modify basal proliferation but it was able to decrease the phytohemagglutinin (PHA)-induced proliferation. Inhibitors of capacitative entry and estrogen receptor antagonist, fulvestrant, but not the NO-synthase inhibitor prevented the effect of delphinidin. In addition, delphinidin reduced actinomycin D-induced apoptosis via a mechanism sensitive to capacitative entry inhibitors. Interestingly, delphinidin decreased the ability of thapsigargin to increase both calcium release and entry. The effect of delphinidin was sensitive to capacitative entry inhibitors and partially prevented by fulvestrant but not by the NO synthase inhibitor. Immunohistochemistry analysis showed that delphinidin reduced membrane translocation of STIM-1, a principal step in capacitative entry activation through « calcium release activated channels » CRAC. Together, these data suggest that delphinidin attenuates T lymphocyte activation via multiple cellular targets on the regulation of calcium signaling via estrogen antagonist sensitive and insensitive pathways. The consequence of which results in inhibition of both proliferation and apoptosis. These effects concur to the protective effect of delphinidin on processes leading to inflammation associated with T lymphocyte activation.

Anthocyanins as substrates for mitochondrial complex I - protective effect against heart ischemic injury.

Anthocyanins, a subclass of flavonoids, are known to protect against myocardial ischemia; however, little is known about their direct, acute effects on mitochondria injured by the ischemic insult. In this study, the effects of delphinidin 3-O-glucoside (Dp3G), cyanidin 3-O-glucoside (Cy3G) and pelargonidin 3-O-glucoside (Pg3G) on the activity of complex I of the mitochondrial respiratory chain were studied in mitochondria isolated from normal rat hearts and rat hearts subjected to ischemia for 45 min. Cy3G and Dp3G increased the activity of complex I, measured in the presence or absence of coenzyme Q1 (CoQ1 ), in ischemia-damaged mitochondria, whereas in nonischemic mitochondria the effect was observed only in the absence of CoQ1 . Dp3G and Cy3G but not Pg3G increased state 3 respiration and ATP synthesis with NADH-dependent substrates in mitochondria after ischemia. The results suggest that certain anthocyanins can act as electron acceptors at complex I, and bypass ischemia-induced inhibition, resulting in increased ATP production after ischemia. This study provides new information on a possible role of certain anthocyanins in the regulation of energy metabolism in mammalian cells.

Abstract 3666: Delphinidin reduces cell proliferation and induces apoptosis of non-small cell lung cancer cells in vitro and in vivo.

Abstract Lung cancer is a major health problem worldwide and accounts for approximately 28% of all cancer-related deaths in the USA. Like most cancers, lung cancer is a conglomeration of diseases of diverse etiology, broadly divided into small-cell lung cancer (SCLC, comprising 20% of lung cancers), and non-small-cell lung cancer (NSCLC, comprising 80% of lung cancers). Lung cancer has proven difficult to control with conventional therapeutic and surgical approaches, and the prognosis is poor with an overall 5-year survival rate of only 15% for NSCLC in the USA. The epidermal growth factor receptor (EGFR) and vascular endothelial growth factor receptor-2 (VEGFR-2) have emerged as two clinically validated targets for NSCLC. The development of dietary agents which exhibit combined EGFR/VEGFR-2 inhibitory activities may be a step forward to improve antitumor efficacy and to broaden application possibilities. In our pursuit for the identification of non-toxic dietary ingredients for prevention and treatment of cancer, we found that delphinidin, an anthocyanidin, present in pigmented fruits and vegetables, is a potent inhibitor of both EGFR and VEGFR-2 in NSCLC cells. In the present study, we determined the effects of delphinidin on cell growth and apoptosis in vitro and on tumor growth and angiogenesis in vivo in NSCLC cells that overexpress EGFR/VEGFR-2. Delphinidin (5-60 μM) treatment of NSCLC cells inhibited the (i) activation of PI3K, and (ii) phosphorylation of AKT and MAPKs. We next determined whether inhibition of EGFR/VEGFR-2 signaling pathways by delphinidin exhibits a growth inhibitory effect. We observed that delphinidin treatment of NCI-H441, SK-MES-1 and A549 cells resulted in a dose-dependent inhibition of cell growth without having any substantial effects on normal human bronchial epithelial cells as assessed by MTT assay. Treatment of NCI-H441 and SK-MES-1 cells with delphindin (5-60 μM) resulted in (i) cleavage of PARP protein, (ii) activation of caspase-3 and -9, (iii) downregulation of anti-apoptotic proteins (Bcl-2, Bcl-xL and Mcl-1), (iv) upregulation of pro-apoptotic proteins (Bax and Bak), and (v) decreased expression of PCNA and cyclin D1. In athymic nude mice implanted with human NSCLC cells (NCI-H441 or SK-MES-1), delphinidin (1-2 mg, i.p. thrice-weekly) treatment caused a significant inhibition of tumor growth. There was also a decrease in the expression of markers for cell proliferation (Ki-67 and PCNA) and angiogenesis (CD31 and VEGF) as well as an induction of apoptosis in tumor tissues treated with delphinidin, when compared with tumor tissues of control mice. Based on these observations, we suggest that delphinidin, alone or as an adjuvant to current therapies, could be useful for the management of NSCLCs, especially those that overexpress EGFR and VEGFR-2. Citation Format: Harish C. Pal, Samriti Sharma, Craig A. Elmets, Farrukh Afaq. Delphinidin reduces cell proliferation and induces apoptosis of non-small cell lung cancer cells in vitro and in vivo. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3666. doi:10.1158/1538-7445.AM2013-3666

Abstract 4981: NADPH oxidase is a novel target of delphinidin to inhibits UVB-induced MMP-1 expression in human dermal fibroblasts

Abstract Ultraviolet (UV) induced ROS production is considered a major cause of photoaging via abnormal expression of matrix metalloproteinase (MMP)-1 and NADPH oxidase (NOX) is a key enzyme of endogenous ROS production. Here, we investigated the anti-photoaging effect of delphidin, a major anthocyanin found in berry, beans, and red wine, and its molecular target in UVB-induced MMP-1 expression using primary cultured human dermal fibroblast. Delphinidin inhibited UVB-induced MMP-1 expression in human dermal fibroblasts. UVB-induced MKK4-JNK1/2, MKK3/6-p38, and MEK-ERK1/2 phosphorylation were suppressed by delphinidin. In addition, quantification of ROS stained with the 2′,7′-dichlorofluorescein diacetate (DCF-DA) dye showed that delphinidin significantly inhibited UVB-induced ROS production. Delphinidin significantly inhibited UVB-induced NOX activity. Interestingly, the inhibitory effect of delphinidin on UVB-induced NOX activity was stronger than apocynin, a pharmaceutical NOX inhibitor. Fractioned cell analysis results using a western blot assay showed that delphinidin strongly suppressed UVB-induced P47phox, a NOX subunit, translocation from the cytosol to the membrane. Delphinidin produced similar effects with apocynin on UVB-induced MMP-1 expression. Pull down assays showed that delphinidin directly bound to P47phox. Collectively, our results suggest that NOX is a potential target of delphinidin in the suppression of UVB-induced MMP-1 expression in human dermal fibroblasts. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4981. doi:1538-7445.AM2012-4981

Abstract 2565: Delphinidin: A dual tyrosine kinase inhibitor of EGFR and VEGFR-2 reduces cell proliferation and induces apoptosis of non-small-cell lung cancer cells

Abstract Lung cancer has proven difficult to control with conventional therapeutic and surgical approaches, and the prognosis is poor with an overall 5-year survival rate of only 15% for non-small-cell lung cancer (NSCLC) in the USA. Over the last decade, the notions of targeting tyrosine kinase receptors for prevention and treatment of cancers have gained considerable attention. The epidermal growth factor receptor (EGFR) and vascular endothelial growth factor receptor-2 (VEGFR-2) have emerged as two clinically validated targets for NSCLC. Dual specific tyrosine kinase receptor inhibitors, which target two different classes of growth factor receptors, represent an elegant way to avoid multiple treatment modalities when used alone and in combination with various anti-signaling agents. The development of dietary agents with combined EGFR/VEGFR-2 inhibitory activities in the same molecule may be a step forward to improve antitumor efficacy and to broaden application possibilities. In our pursuit for identification of non-toxic dietary ingredients for prevention and treatment of cancer, we found that delphinidin, an anthocyanidin, present in pigmented fruits (such as pomegranate, berries, and dark grapes) and vegetables (such as egg plant, tomato, carrot and red onion), is a potent inhibitor of EGFR and VEGFR-2 in NSCLC cells. Employing EGFR and VEGFR-2 over expressing NSCLC cells (NCI-H441 and SK-MES-1); we evaluated the effect of delphinidin on EGFR/VEGFR-2 downstream signaling pathways. Delphinidin (5-60 μM) treatment of both NCI-H441 and SK-MES-1 cells inhibited the (i) activation of PI3K, and (ii) phosphorylation of AKT and mitogen activated protein kinases (such as ERK1/2, JNK1/2 and p38). We next determined whether inhibition of EGFR/VEGFR-2 signaling pathways by delphinidin exhibits growth inhibitory effect. We observed that delphinidin treatment of NCI-H441, SK-MES-1 and A-549 cells resulted in a dose-dependent inhibition of cell growth without having any substantial effect on normal human bronchial epithelial cells as assessed by an MTT assay. Treatment of NCI-H441 and SK-MES-1 cells with delphindin (5-60 μM) resulted in (i) cleavage of PARP protein, (ii) activation of caspase-3 and-9, (iii) downregulation of anti-apoptotic proteins (Bcl-2, Bcl-xL and Mcl-1), (iv) upregulation of pro-apoptotic proteins (Bax, Bid and Bak), and (v) decrease expression of PCNA and cyclin D1. In addition, we found that delphinidin was effective in reducing the invasive potential of NSCLC cells. This study identifies an abundant fruits and vegetables based anthocyanidin delphinidin as an effective blocker of EGFR and VEGFR-2 in lung cancer cells. Based on these observations, we suggest that delphinidin, alone or as an adjuvant to current therapies, could be useful for the management of NSCLC. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2565. doi:1538-7445.AM2012-2565

Abstract LB-426: Delphinidin suppresses ultraviolet B-induced cyclooxygenases-2 expression through inhibition of MAPKK4 and PI-3 kinase

Abstract Cyclooxygenase-2 (COX-2), a key mediator of inflammation, and its product, prostaglandin E2 (PGE2), enhance carcinogenesis, particularly in skin. Ultraviolet (UV) B is the most carcinogenic component of solar irradiation, and a crucial role of COX-2 in UVB-mediated skin carcinogenesis has been reported. Here, we investigated the effects of delphinidin, an abundant dietary anthocyanin, on UVB-induced COX-2 upregulation and the underlying molecular mechanism. We found that delphinidin suppressed UVB-induced COX-2 expression in JB6 P+ mouse epidermal cells. COX-2 promoter activity and PGE2 production were also suppressed by delphinidin treatment within non-cytotoxic concentrations. Activator protein-1 and nuclear factor- B, crucial transcription factors involved in COX-2 expression, were activated by UVB and delphinidin abolished this activation. UVB-induced phosphorylation of c-Jun N-terminal kinase, p38 kinase and Akt was inhibited by delphinidin. The activities of mitogenactivated protein kinase kinase (MAPKK) 4 and phosphatidylinositol-3 kinase (PI-3K) were inhibited markedly by delphinidin. A pull-down assay using delphinidin-Sepharose beads revealed that delphinidin binds directly with MAPKK4 or PI-3K in a manner that was competitive with adenosine triphosphate. Moreover, in vivo investigations using mouse skin revealed that the upregulation of COX-2 expression, MAPKK4 activity and PI-3K activity induced by UVB was abolished with delphinidin treatment. Collectively, our results demonstrated that delphinidin targets MAPKK4 and PI-3K directly to suppress COX-2 overexpression, suggesting a potential protective role for delphinidin against UVB-mediated skin carcinogenesis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-426.