bluuebunny/crossref_metadata_2025 huggingface dataset:
Curcumin Inhibits Lymphangiogenesis in vitro and in vivo through Suppressing VEGF Receptor Signaling
Authors: Wang Weicang
Published: November 2023

Curcumin, a major curcuminoid of turmeric, has been shown to inhibit tumor growth and metastasis; while the underlying mechanisms are not well understood. Here we investigated the effects of curcumin on lymphangiogenesis (de novo formation of lymphatic vessels), which is a critical process implicated in tumor metastasis. Our results showed that curcumin inhibited lymphangiogenesis in vitro and in vivo, through suppressing vascular endothelial growth factor (VEGF) receptor signaling. Curcumin inhibited VEGF‐C‐induced infiltration of LYVE‐1‐positive lymphatic endothelial cells in a Matrigel plug assay in mice, demonstrating its anti‐lymphangiogenic effects in vivo. In human dermal microvascular lymphatic endothelial cells (HMVEC‐dLy), curcumin inhibited multiple lymphangiogenic responses, including cell proliferation, migration and capillary‐like tube formation in a dose‐dependent manner. Curcumin inhibited lymphangiogenesis, through suppressing the expressions of VEGF receptors (VEGFR2 and VEGFR3), and phosphorylation of downstream signaling proteins such as ERK and FAK. Finally, curcumin sulfate and curcumin glucuronide, two major metabolites of curcumin, had little effect on lymphangiogenic responses in HMVEC‐dLy cells. Together, our results demonstrate that curcumin inhibited lymphangiogenesis in vitro and in vivo through suppressing VEGF receptor signaling, suggesting a novel mechanism for the anti‐metastatic effects of curcumin.
Terpenoid‐enriched Loquat Leaf Extract Controls the Growth and Survival of Pancreatic Cancer Cell and Acts on Glucose Metabolsim to Alter Pentose Phosphate Pathway
Authors: Lu Qing‐Yi , Yang Jeiping , Go Vay‐Liang , Li Zhaoping , Lee Wai‐Nang
Published: November 2023

Background

Eriobotrya japonica (Thunb) Lindl. (Rosaceae), or loquat, is a fruit native to China and Japan. Loquat leaves have been used for the treatment of diabetes mellitus, chronic bronchitis, coughs and skin diseases in traditional Chinese Medicine. The biological active constituents of leaf included terpenoids, which have shown to exhibit anti‐hyperglycemia effect and to promote insulin release by stimulating pancreatic beta‐cells in animal models.

Aim

The present study investigated novel anti‐tumor activities and the alteration of cellular metabolism by a loquat leaf extract (LLE).

Method

The terpenoid‐enriched extract was prepared by the extraction of dried leaves growing in Southern California with 75% aqueous ethanol. Mia PaCa‐2 pancreatic adenocarcinoma cells were cultured for 48 hours in presence of various concentrations of LLE and its major terpene constituent ursolic acid (UA), and the effect of LLE and UA on the growth and apoptosis of tumor cells was determined. Using 50% [1, 2 13C2]‐glucose as a single precursor metabolic tracer, extracellular lactate production as well as intracellular de novo fatty acid palmitate synthesis, acetyl‐CoA and ribonucleotide ribose were measured.

Results

Our results show that LLE and UA dose‐dependently induce growth suppression and apoptosis, and inhibit phosphor‐AMPK expression. LLE (50 mg/mL) reduces the contribution of glucose to the lactate production and inhibits fatty acid, acetyl‐CoA (P < 0.01) and ribonucleotide ribose (P < 0.01) syntheses. Mass isotopomer analysis also reveals that the treatment of LLE results in marked decrease in pentose cycle flux through the oxidative relative to the non‐oxidative pathway (P < 0.01). Similar results were observed by UA (50 mM) and metformin (1 mM) treatment with the exception that metformin increases lactate production (P < 0.05).

Our study demonstrates that LLE alters glycolysis‐pentose phosphate pathway and controls glucose‐utilizing biomolecule syntheses essential to tumor cell growth and survival.

Support or Funding Information

Supported by the Hirshberg Foundation for Pancreatic Cancer Research
Apiaceous Vegetable Intake Modulates Expression of DNA Damage Signaling Pathway Genes and miRNA in Wistar Rats
Authors: Kim Jae Kyeom , Gallaher Daniel D , Trudo Sabrina P
Published: November 2023

2‐Amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine (PhIP) is the most mass abundant heterocyclic aromatic amine in foods and is a colon‐specific carcinogen. We recently showed that PhIP‐DNA adducts in rat colon were reduced by 20% when the diet was supplemented with apiaceous vegetables (i.e., celery and parsnips) at a modest dose (21%, wt/wt) equivalent to a human intake of 1 cup/day (based on a 2,000 kcal diet). The mechanism of action, however, was not through a commonly hypothesized effect, inhibition of pro‐carcinogen activation. Thus, here we examine the effects of apiaceous vegetable intake on promising alternative mechanisms of dietary chemoprevention: 1) N‐methyltransferase (NMT) protein expression, 2) gene expression of PhIP transporters, 3) expression of genes related to DNA damage signaling pathways, and 4) changes in the global microRNA (miRNA) profile in rat colon. Male Wistar rats fed either the American Institute for Nutrition (AIN)‐93G diet (n=6), or the AIN‐93G supplemented with apiaceous vegetables (API; n=7), received a single intraperitoneal injection of PhIP (10 mg PhIP/kg body weight) after 6 days of feeding. A negative control group was fed AIN‐93G and injected with DMSO (n=5). No difference in hepatic NMT protein expression was noted amongst the groups. There was a trend for increased colonic mRNA expression of p‐glycoprotein, a minor PhIP efflux transporter (1.5‐fold; P=0.075); mRNA expression of other colonic PhIP efflux transporters, BCRP and MRP2, was not changed. PCR array analysis revealed that mRNA expression of Pole and Rad18, involved in DNA repair pathways and induced in response to DNA damage, was reduced by API (1.3‐fold for both; P<0.05). Also, in the global miRNA profiling analysis, 11 out of 421 miRNAs were either elevated or suppressed in response to apiaceous vegetable supplementation. In particular, miR‐19b‐3p and miR‐29b‐3p were increased by API diet (2.8‐ and 2.2‐fold increase, respectively, P<0.05). Predictions by Ingenuity Pathway Analysis suggest that these two miRNAs target BMPR2 and CCND1, among others. Some evidence suggests BMPR2 may be oncogenic, and there are reports of a positive correlation between CCND1 overexpression and cancer onset. In conclusion, apiaceous vegetable intake had negligible effects on NMT protein and PhIP transporter gene expression, but did modify genes and miRNA with potential chemopreventive roles. Further investigations are warranted regarding the impact of apiaceous‐mediated modulation of Pole and Rad18, and the net downstream effects of miRNA modulation as pertains to colon carcinogenesis.

Support or Funding Information

University of Minnesota Masonic Cancer Center
Synergystic Interaction of Common Bean (Phaseolus vulgaris L.) Peptides and Oxaliplatin on Proliferation of HCT116 Colorectal Cancer Cells through Mitochondrial Depolarization and DNA Damage
Authors: Luna‐Vital Diego‐Armando , DE MEJIA ELVIRA GONZALEZ , Loarca‐Pina Guadalupe
Published: November 2023

The objectives of this study were to evaluate the antiproliferative effect of common bean (Phaseolus vulgaris L.) peptides on HCT116 human colorectal cancer cells and their interaction with oxaliplatin; furthermore, to determine their potential mechanisms of action. Pure peptides GLTSK, LSGNK, GEGSGA, MTEEY and MPACGS and the peptides in the non‐digestible fraction from common bean cultivars Azufrado Higuera (AH) and Bayo Madero (BM) were tested for cytotoxic effects on HCT116, and also on CCD‐33Co human normal colon cells. None of the peptides were toxic to normal cells at concentrations up to 1 mM. GLTSK (IC50=134.6 μM). On the contrary, according to MTS assay, GEGSGA (IC50=156.7 μM) exerted antiproliferative effects on HCT116 cells.. Based on the combination indexes (γ) of the isobolographic analysis, GLTSK (γ=0.64) and GEGSGA (γ=0.78) interacted synergistically (p < 0.05) with oxaliplatin inhibiting HCT116 cells. Using flow cytometry, it was determined that GLTSK caused loss of mitochondrial membrane potential (Δψm) (16% of cells), increased (p < 0.05) in intracellular reactive oxygen species (ROS) (12.1‐fold compared with untreated cells), cell cycle arrest in G2 (54% of cells) and induced apoptosis (33% of cells). GEGSGA had a limited effect on Δψm and intracellular ROS, but caused arrest in G1 phase (64% of cell population) and promoted increase (p < 0.05) in p53 (290%) and cleavage of PARP (184%) compared to untreated cells, promoting DNA damage. The different mechanisms of action were further established by confocal microscopy. An increase (p < 0.05) in the expression and activation of nuclear p53 (145% of fluorescence intensity/μm2) was observed in response to GEGSGA treatment compared to untreated cells in a time‐frame of 3 to 8 h. Moreover, GLTSK depolarized the cells (38% decrease of fluorescence intensity/μm2) and increased (p < 0.05) intracellular ROS (180% of fluorescence intensity/μm2). In addition, peptide fractions from bean cultivar Azufrado Higuera interacted synergistically with oxaliplatin inducing apoptosis (49% of cells), loss of Δψm (21.7% of cells) and increase of intracellular ROS (13.1‐fold compared to untreated cells). This is the first report demonstrating that dietary peptides from common bean interact synergistically with oxaliplatin inhibiting HCT116 human cancer cells. These results support the hypothesis that physicochemical properties of peptides would determine their mechanism of action.

Support or Funding Information

Basic Science, National Council for Science and Technology, Mexico CONACyT‐grant number 220924
1α,25‐Dihydroxyvitamin D3 Inhibits De Novo Fatty Acid Synthesis and Metastatic Capability of Breast Cancer Cells
Authors: Wilmanski Tomasz , Barnard Alle , Donkin Shawn , Parikh Mukti , Buhman Kimberly , Kirshner Julia , Burgess John , Teegarden Dorothy
Published: November 2023

Breast cancer continues to be the second most commonly diagnosed cancer in American women. Although treatment of localized breast cancer has been effective with five year survival reaching 99%, survival rates drastically decrease to 24% once the tumor has metastasized to distant secondary sites. The most common site of breast cancer metastasis is bone and both higher intracellular lipid accumulation and upregulated de novo fatty acid synthesis correlate with greater breast cell metastatic capability. Therefore, strategies targeting breast cancer cell lipid metabolism may be successful in inhibiting breast to bone metastasis. Here, we demonstrate that the active vitamin D metabolite, 1α,25‐dihydroxyvitamin D (1,25(OH)2D), inhibits the metastatic capability of MCF10CA1a (30% of vehicle) and MDA‐MB‐231 breast epithelial cells (80% of vehicle) in an in vitro 3D (rMET) model of breast to bone metastasis. In addition, we show that 1,25(OH)2D regulates breast cancer cell lipid metabolism by inhibiting de novo fatty acid synthesis from 13C[U]‐Glucose (48%±5.5 relative to vehicle) and decreasing neutral lipid accumulation (BODIPY staining) (56%±7.4 relative to vehicle) at 5 days of treatment. In our studies investigating the mechanisms of 1,25(OH)2D mediated regulation of lipid metabolism, we identified the mitochondrial enzyme pyruvate carboxylase (PC) as a primary target of 1,25(OH)2D. Treatment with 1,25(OH)2D decreased both PC mRNA (58%±7 relative to vehicle) and protein (54%±12 relative to vehicle) expression at 5 days. Consistent with these results, de novo fatty acid synthesis from acetate, the immediate substrate for fatty acid synthesis downstream of glucose, was not changed with 1,25(OH)2D, supporting a role of PC in 1,25(OH)2D mediated inhibition of fatty acid synthesis upstream of lipogenic enzymes. Thus, our studies demonstrate that 1,25(OH)2D decreases fatty acid accumulation and metastatic capability, potentially through downregulation of PC.

Support or Funding Information

This work was supported by a Project Development Team within the ICTSI NIH/NCRR Grant Number UL1TR001108, as well as the National Institutes of Health, National Cancer Institute R25CA128770 (D. Teegarden) Cancer Prevention Internship Program administered by the Oncological Sciences Center and the Discovery Learning Research Center at Purdue University.
Pyrogallol, a microbial metabolites from mango tannins (Mangifera Indica L.) suppresses breast cancer ductal carcinoma in situ proliferation in both in vitro and in vivo
Authors: Mertens‐Talcott Susanne U , Kim Hyemee , Barnes Ryan , Talcott Stephen T , Nemec Matthew
Published: November 2023

It is estimated that 1 in 10 newly diagnosed cancer cases will be female breast cancer, and therefore there is an ongoing need to research novel treatment options. Polyphenolics are secondary plant metabolites that have been shown to have anti‐cancer effects in multiple cancer models without the deleterious side effects of conventional small molecules. This research, focused on the anti‐proliferative activities of pyrogallol, an intestinal microbial metabolite of mango polyphenols. Mice were xenographed with MCF10DCIS.COM cells in order to determine the anti‐proliferative of pyrogallol on ductal carcinoma in situ breast cancer (DCIS). Treatment animals received either 0.2mg/day or 0.8mg/day pyrogallol, and both treatment groups had significantly decreased tumor volumes compared to the control animals after 4 weeks of treatment. Plasma and tumor tissue was assessed in order to determine in pyrogallol or its metabolites were present in circulation or at the site of action. Additionally, biomarkers along the AMPK‐mTOR signaling axis were affected at mRNA, protein and phosphorylated protein levels. These results suggest that anti‐proliferative activities of mango polyphenols may be mediated by pyrogallol as a central bioactive metabolite that is systemically absorbed. Findings indicate that a diet rich in gallotannins that are microbially metabolized to pyrogallol may prevent the advancement of DCIS breast cancer.

Support or Funding Information

Mango National Board
Comparing Gene Expression Between Resveratrol Sensitive and Insensitive Cells Using RNA‐Sequencing Methods
Authors: Cranney Caleb Webster , Kenealey Jason
Published: November 2023

Cancer treatments, while steadily improving in quality and effectiveness, continue to depend primarily on compounds toxic to non‐cancerous cells. These treatments, while demonstrating effectiveness against specific strains of cancer, often lead to debilitating and degenerate human disease. Resveratrol, a naturally occurring polyphenol found in the skin and seeds of red grapes, has been demonstrated to have anti‐cancerous properties. While much research has been done the general chemotherapeutic effect of resveratrol, little has been determined about the precise mechanism through which resveratrol specifically targets cancerous cells. By studying these mechanisms, one could perhaps learn to mimic or enhance it's nontoxic chemotherapeutic qualities in cancer treatments. We analyzed gene expression data between cancer cells with and without resveratrol treatment (focusing on both coding and noncoding RNA) to pinpoint with greater accuracy the biochemical pathways involved in this unique chemotherapeutic. To examine this, RNA sequencing methods were performed on MDA‐MB‐231 breast cancer cells on samples with and without resveratrol treatment. We applied RNA sequencing methods in a similar manner to a strain of the same cell line that developed an insensitivity to resveratrol. This cell line was developed through incrementally increasing resveratrol concentrations in the media over 8 months. The data was run through a Gene Set Omic Analysis (GSOA) program developed by bioinformatics professors at BYU. We have identified pathways that are modulated in response to resveratrol that help elucidate the chemotherapeutic mechanism of resveratrol. Studying impacted biochemical pathways identified by the analysis can reveal nontoxic methods of cancer treatment.

Support or Funding Information

Simmons Center for Cancer Research Undergraduate Research Fellowship, BYU Life Sciences Start‐up Grant
Carotenoid profile and antioxidant activity of tortillas elaborated from yellow creole maize (Zea mays L.) by conventional nixtamalization process
Authors: Mora‐Rochin Saraid , Bañuelos Ana Belinda Corrales , Serrano Diego Misael Angulo , Rodriguez Edith Oliva Cuevas , Moreno Cuauhtemoc Reyes , Milan‐Carrillo Jorge
Published: November 2023

Maize (Zea mays L.) is a widely consumed cereal in Mexico and Central America, mainly in the form of tortillas. Creole maize grains have multiple colors provided by two large families of pigments: 1) phenolics and 2) carotenoids, which are the most abundant in maize. Carotenoids are subject to many chemical changes induced by different processing conditions used in the food industry. The main process at which maize is subject to prepare tortillas is nixtamalization. This process may generate lossess of bioactive compounds due to the thermal alkaline cooking, however, it's been shown that the remaining compounds have a better bioavailability. The objectives of the present study were to evaluate the effect of traditional nixtamalization process and the profile levels of carotenoids, total phenolic content, hydrophilic (ORACH, ABTS and DPPH) and lipophilic (ORACL) antioxidant activity on four different yellow (Y) creole maize (264, 317, 320 and 502), and processed into tortillas. The results showed a 50% decreased in the total phenolic content, and hydrophilic antioxidant activity (ORAC, ABTS and DPPH) when nixtamalized tortilla (NT) was analyzed. This could be related with the concentrations found in tortillas made from creole maizes, which are higher than those reported by other authors. Among the pigmented genotypes, NT made from Y‐502 maize showed the highest total phenolic content (147.7 mg GAE/100 g DM), and antioxidant activity using the ORACH (6,467 μmol TE/100 g DM), DPPH (1,385 μmol TE/100 g DM) and ABTS (4,250 μmol TE/100 g DM) methods. Additionally, the same genotype showed the highest concentration of total carotenoids and lipophilic antioxidant capacity (531.9 μg Lutein/100 g DM and 13,492 μmol TE/100 g DM). On the other hand, carotenoid profiles analysis showed 64% increased in Lutein when NT made from Y‐317 maize were used. However, when Lutein and Zeaxanthin were analyzed in raw grain and NT, higher concentrations of carotenoids were found. The present study demonstrated that nixtamalization of yellow creole maize from Sinaloa, not only retain bioactive compounds and has antioxidant capacity, but also increases the concentration of bioactive compounds.

Support or Funding Information

PROFAPI‐UAS‐2014‐137 AND CONACYT CIENCIA BASICA (168279)
Efficacy of Chewing Gum as a Delivery System for Water and Fat Soluble Vitamins
Authors: Khoo Weslie Yu Heng , Lambert Joshua D
Published: November 2023

Chewing gum is a popular product among consumers and chewing gums containing functional ingredients including botanical extracts and vitamins are commercially‐available. In the present study, we examined the release kinetics and plasma bioavailability of a panel of water‐soluble and fat‐soluble vitamins from two commercially available vitamin supplemented gums. We have recruited 15 healthy subjects (age: 22–52 yrs, 60% women) and employed a single‐blind, placebo‐controlled cross‐over study design. Time‐dependent changes in saliva and plasma levels of vitamins were determined by high‐performance liquid chromatography using both ultraviolet light and electrochemical detection methods. In general, the component water‐ and fat‐soluble vitamins in the gums were released from the matrix into the saliva, and the levels in the plasma were elevated compared to levels following the placebo. We found that the release of water‐soluble ascorbic acid, niacinamide, and pyroxidine from the gum into the saliva during chewing was 52.1–62.1%, 76.8–80.6% and 40.0–70.4% of its available content, respectively. The release of fat‐soluble dl‐alpha‐tocopheryl acetate into the saliva during chewing was 30.4–44.0% of the available content. Furthermore, there was a near‐complete release of fat‐soluble retinyl palmitate content from the gum into the saliva. Plasma ascorbic acid and niacinamide levels were increased by 78.8–82.8% and 215.1–273.4%, respectively, in vitamin‐supplemented chewing gums compared to baseline. Analysis of plasma retinol and dl‐alpha‐tocopherol are still ongoing. To the best of our knowledge, this is the first report of the efficacy of chewing gum to deliver vitamins to human subjects. Further studies in deficient subjects are needed to determine whether chronic use of vitamin‐supplemented chewing gum is an effective approach to correct deficiency.

Support or Funding Information

Vitaball Inc.
Enhanced Solution Concentration of Quercetin in Amorphous Solid Dispersions
Authors: Gilley Andrew , Arca Hale Cigdem , Nichols Brittany , Edgar Kevin , Neilson Andrew Peter
Published: November 2023

The flavonol quercetin is present in foods such as apples and onions and has potential beneficial bioactivities against cardiovascular disease, obesity, and oxidative stress. However, the benefits of quercetin are limited by poor bioavailability, primarily attributed to its crystallinity (resulting in low aqueous solubility) and extensive Phase‐II/III metabolism. Increasing solution concentrations of quercetin in the intestinal lumen has the potential to increase its diffusion across the unstirred water layer, thus improving absorption. Improving the bioavailability of quercetin is critical for enhancing its efficacy in vivo. Various strategies have been employed to improve quercetin dissolution in the intestinal lumen. One promising approach is to incorporate quercetin into amorphous solid dispersions (ASD). Cellulosic polymers with carboxylic acid functionality disperse quercetin, eliminating crystallinity, and selectively release quercetin at intestinal pH due to electrostatic repulsion of deprotonated acid groups. We assessed the utility of commercially available and novel cellulose derivatives for improving in vitro dissolution properties of quercetin, measured by the area under the concentration/time curve (AUC). Quercetin was dispersed via spray drying with novel cellulose derivatives [cellulose acetate suberate (CASub) and cellulose acetate adipate propionate (CAAdP)], and commercially available derivatives [hydroxypropylmethyl cellulose (HPMCAS) and carboxycellulose acetate butyrate (CCAB)]. Amorphous structure was confirmed by X‐ray diffraction. In vitro dissolution was performed at gastric and small intestinal pH (1.2 and 6.8, respectively). Crystalline quercetin showed poor solubility at both gastric and intestinal pH. At pH 1.2, amorphous quercetin in all cellulose derivatives showed poor dissolution, as expected. At pH 6.8, 10% (w/w) amorphous quercetin in CCAB provided significantly enhanced quercetin solution concentrations compared to crystalline quercetin and 25% and 50% amorphous quercetin; thus, 10% quercetin was employed for the other polymers. Amorphous quercetin (10%) in HPMCAS provided enhanced dissolution profile for quercetin at intestinal pH, substantially increasing its AUC compared to quercetin dispersed in CCAB. These results provide promising, novel opportunities for increasing the bioavailability of quercetin in vivo, and support the use of HPMCAS to achieve this. Furthermore, in vivo bioactivity studies are warranted to assess the ability of ASD to improve upon the bioactivity of crystalline quercetin.

Support or Funding Information

Funding provided by the Institute for Critical Technologies and Applied Science at Virginia Tech.
Mango polyphenols (Mangifera Indica L.) and their microbial metabolites suppress adipogenesis and fat accumulation by mediating AMPK signaling pathways in 3T3L‐1 adipocytes
Authors: Fang Chuo , Kim Hyemee , Talcott Stephen T , Mertens‐Talcott Susanne U
Published: November 2023

The prevalence of obesity and obesity‐related diseases has increased dramatically in the United States over the last decades. Obesity is characterized by an excessive accumulation of triglycerides in adipose tissue and other metabolic tissues associated with abnormal levels of adipocytokines and inflammatory cytokines. Previously, polyphenols were shown to have anti‐lipogenic and anti‐inflammatory properties. The AMPK‐signaling pathway is involved in the regulation of adipogenesis and inflammation and has been considered a therapeutic target in the prevention and treatment of obesity. In this study, mango and its microbial metabolites, were tested to determine their roles in regulating lipid metabolism and adipogenesis via the activation of AMPK in differentiated 3T3‐L1 adipocytes. Treatments inhibited cell proliferation in a dose‐dependent manner. The activation of AMPK pathways by mango polyphenols suppressed cell differentiation via modulation of the mitotic clonal expansion phase, and reduced the mRNA levels of PPARγ, C/EBPα and SRBEP‐1c. It also exerted an inhibitory effect on the adipogenic gene expression of FAS, ACC, as well as a stimulative effect on lipolytic gene expression of HSL, ATGL, and fatty acid transporters. The levels of NF‐κB and pro‐inflammatory cytokines such as IL‐1β, IL‐6 and TNF‐α were reduced by our mango and its metabolites. These results suggest that a diet abundant in mango might be helpful in the prevention of obesity and obesity‐related diseases.
Inhibition of high glucose‐induced diabetic retinal angiogenesis by dietary compound chrysin
Authors: Kang MinGyeong , Kang Young‐Hee
Published: November 2023

Diabetic retinopathy (DR) has been implicated in patients suffering from chronic diabetes mellitus as pathological complications, characterized by diabetic retinal edema and abnormal retinal blood vessel outgrowth leading to diabetic vision loss. Chrysin is a natural flavonoid found in herb and honeycomb and possesses various bioactive properties. This study investigated to determine protective effects of chrysin on retinal angiogenesis caused by diabetic episode. Human retinal microvascular endothelial cells were incubated in media containing 27.5 mM mannitol (+5.5 mM glucose) or 33 mM glucose for 2 d in the presence of non‐toxic chrysin at 1–20 μM. High glucose upregulated the induction of the angiogenic markers of hypoxia inducible factor (HIF)‐1α and vascular endothelial growth factor (VEGF). In contrast, chrysin attenuated the induction of these angiogenic marker proteins. In the in vivo animal model of db/db mice, oral administration of 10 mg/kg chrysin decreased the retinal induction of HIF‐1α and VEGF and blocked retinal edema in db/db mice. These result demonstrated that chrysin inhibited retinal angiogenesis and edema retinal by attenuating the induction of angiogenic factors under diabetic circumstances. Therefore, chrysin may be a therapeutic agent in antagonizing diabetic retinopathy.

Support or Funding Information

This study was supported by National Research Foundation of Korea (2015R1A2A2A01006666) and Brain Korea 21 plus program by Ministry of Education of Korea (22A20130012421).
Gossypetin suppresses acidification‐related protein expression and lysosomal enzymes secretion in RANKL‐mediated osteoclasts
Authors: Antika Lucia Dwi , Lee Eun‐Jung , Kang Young‐Hee
Published: November 2023

The imbalance between osteoblastic bone formation and osteoclastic bone resorption in the bone marrow microenvironments leads to osteolytic bone diseases including osteoporosis. Osteoclast‐mediated bone resorption is accomplished by delivering proton (H+) and secreting lysosomal formation of cathepsin K into acidic resorption lacunae. Gossypetin, a flavonoid isolated from Hibiscus species has been shown to possess anti‐microbial and anti‐atherosclerotic activities. The current study investigated the anti‐osteoporotic effects of 1–20 μM gossypetin on suppressing bone resorption in 50 ng/ml receptor activator of nuclear factor κB‐ligand (RANKL)‐differentiated Raw 264.7 murine macrophages for 5 days. When gossypetin was supplemented to RANKL‐induced osteoclasts, the number of multinucleated osteoclasts was reduced and the area of the resorption pits was diminished in a dose‐dependent manner, evidenced by tartrate‐resistance acid phosphatase activity and bone resorption kit assay. Western blot and PCR analysis showed that 20 μM gossypetin abrogated the induction of carbonic anhydrase II, vacuolar‐type H+‐ATPase and chloride channel‐7, all responsible for the bone acidification. In addition, gossypetin suppressed organic matrix degradation of mature osteoclasts through down‐regulating lysosomal enzymes secretion containing cathepsin K and matrix metalloproteinase‐9, indicating the decreased bone resorption activity. These results demonstrated that gossypetin might be a potential agent targeting against osteoclastogenic resorption.

Support or Funding Information

This study was supported by Brain Korea 21 plus program by Ministry of Education of Korea (22A20130012421).
Astragalin attenuates allergic airway inflammation by modulating macrophage infiltration in asthmatic mice
Authors: Kim Yun‐Ho , Choi Yean Jung , Kang Young‐Hee
Published: November 2023

Pulmonary fibrosis and destruction are likely to be the main contributors to pathophysiological lung dysfunction, which may be attributed to any subsequent development of emphysema. We recently reported that astragalin allayed oxidative stress‐associated pulmonary fibrosis through disturbing airway epithelial mesenchymal transition and epithelial autophagic stress. This study attempted to identify how airway inflammation and remodeling led to the failure of lung function and how astragalin affected airway and pulmonary remodeling in the progression of inflammatory lung diseases including asthma, emphysema and pulmonary fibrosis. We found that astragalin attenuated pro‐inflammatory responses through diminishing the MCP‐1 secretion and the ICAM‐1 expression in 20 μM H2O2‐exposed airway epithelial BEAS‐2B cells. Oral administration of astragalin suppressed the induction of MCP‐1, CD11b(+) and F4/80(+) in OVA‐sensitized mouse airways and lungs. Additionally, mast cells were infiltrated in the airway epithelium in OVA‐challenged mice, and such infiltration was blocked by supplementing 20 mg/kg astragalin in sensitized‐mice. Our results demonstrated that astragalin may ameliorate pulmonary inflammation and remodeling by suppressing the production of several inflammatory mediators.

Support or Funding Information

This study was supported by Brain Korea 21 plus program by Ministry of Education of Korea (22A20130012421).
Lycopene Induces Apoptosis in Pancreatic Cancer Cells
Authors: Jeong Yoonseon , Lim Joo Weon , Kim Hyeyoung
Published: November 2023

Pancreatic cancer is one of the most fatal human cancers, which has the lowest 5‐year survival rate among all cancer types. Lycopene, a major carotenoid component in tomatoes, has a potential anticancer activity in many types of cancers. The inhibitors of apoptosis (IAPs) including survivin, cellular inhibitor of apoptosis 1(cIAP‐1) and cellular inhibitor of apoptosis 2(cIAP‐2) are a group of anti‐apoptotic factors in the apoptotic pathway that cause cancer cells become insensitive to apoptotic stimulation. The purpose of this study is to investigate the anti‐cancer mechanism of lycopene by determining the expression levels of inhibitors of apoptosis in human pancreatic cancer cells, PANC‐1. The cells were treated with various concentrations of lycopene or caspase‐3 inhibitor Z‐VAD_FMK. Cell viability was examined by MTT assay. Expressions of survivin, cIAP‐1 and cIAP‐2 were determined by real‐time PCR. We found that the cell viability was decreased by lycopene. A caspase inhibitor Z‐VAD_FMK suppressed lycopene‐induced cell death. In addition, lycopene decreased mRNA expression of survivin, cIAP1 and cIAP2 in pancreatic cancer cells. These results indicate that lycopene induces the apoptosis of pancreatic cancer cells by suppressing the expression of survivin, cIAP1 and cIAP2. Lycopene may be a promising therapeutic agent for human pancreatic cancer.
Activation of AMP Kinase by Zyflamend, a Well‐Defined Blend of Herbal Extracts
Authors: MacDonald Amber Frances , Han Anna Frances , Donohoe Dallas , Whelan Jay
Published: November 2023

Zyflamend, a select blend of 10 herbal extracts, has been shown to inhibit prostate cancer (PCa) in vitro and in vivo at human equivalent doses. Zyflamend's tumor suppressive effects are partly due to the phosphorylation and activation of 5′ adenosine monophosphate‐activated protein kinase (AMPK), a master energy sensor of the cell that restores homeostasis in response to metabolic stress. AMPK activation by Zyflamend inhibits lipogenesis and enhances mitochondrial fatty acid oxidation, depriving proliferating tumor cells of necessary fatty acids for the generation of new membranes (Zhao et al, 2015). However, the mechanism of action whereby Zyflamend phosphorylates AMPK (pAMPK) is unknown. Currently, the literature has identified four kinases that phosphorylate the catalytic subunit of AMPK at Thr172, thereby increasing activity. Two of these kinases that most likely activate AMPK in PCa are liver kinase B1 (LKB1) and calcium/calmodulin‐dependent protein kinase kinase β (CaMKKβ). First, inactivation of LKB1 is associated with a variety of cancers, including reduced expression in PCa tissue. In addition, knockdown of LKB1 promotes the proliferation of PCa. Second, CaMKKβ has been negatively associated with castrate‐resistant PCa, where decreased expression increases proliferation and over expression reduces tumor growth in vivo. Therefore, the objective of this study was to determine the upstream kinase of AMPK activation by Zyflamend in castrate‐resistant PCa cells. CWR22Rv1, a castrate‐resistant PCa cell line, expresses LKB1 and CaMKKβ. When cells were treated with chemical inhibitors of LKB1 (radicicol) or CaMKKβ (STO‐609), pAMPK was reduced. Treatment with Zyflamend in the presence of radicicol failed to completely restore pAMPK at Thr172, while pAMPK was restored to the Zyflamend‐stimulated levels in the presence of STO‐609. These data suggest that Zyflamend‐stimulated pAMPK is partially dependent upon LKB1, but independent of CaMKKβ activity.

Support or Funding Information

Supported by the Tennessee Agricultural Experiment Station Hatch grant #TEN00441
Genetic Background Determines Anthocyanin Effects on Glutathione Redox Balance
Authors: Norris Katie M , Pazdro Robert
Published: November 2023

Background

Anthocyanins are secondary plant metabolites that provide the blue, red, and purple color of many plants. In recent years, these compounds have generated considerable interest among biomedical researchers due to their reported health benefits. It has been proposed that commonly consumed anthocyanins, such as cyandin‐3‐O‐β‐glucoside (C3G), confer cellular protection by increasing biosynthesis of glutathione (GSH), an endogenous antioxidant. However, it is unclear whether anthocyanin‐induced health effects are subject to genetic control. In this project, we tested the hypothesis that genetic background regulates anthocyanin‐induced alterations in GSH levels and redox balance (GSH/GSSG).

Methods

Female mice representing five genetically diverse inbred strains (A/J, 129S1/SvImJ, CAST/EiJ, C57BL/6J, and NOD/ShiLtJ) were assigned to a control (n=5/strain) or 100 mg/kg C3G diet (n=5/strain) for six weeks. Liver, kidney, heart, pancreas, and brain were collected, and levels of GSH and GSSG were quantified by high‐performance liquid chromatography (HPLC). Total glutathione was calculated as GSH + 2GSSG, and redox balance was assessed by the GSH/GSSG ratio.

Results

C3G treatment increased GSH/GSSG in the livers (p = 0.041) and hearts (p = 0.028) of CAST/EiJ mice. C3G increased renal GSH levels (p = 0.01), increased total renal glutathione (p = 0.014), and decreased pancreatic GSSG levels (p = 0.021) in this strain as well. C3G‐treated C57BL/6J mice exhibited increased GSH/GSSG ratios in the pancreas (p = 0.042) only. NOD/ShiLtJ mice exhibited no differences between control and C3G treatment in any of the tissues, and no significant differences were found with C3G treatment in the brain in any of the five strains. Surprisingly, C3G negatively affected GSH levels in two strains. A/J mice fed a high C3G diet exhibited lower hepatic GSH concentrations (p = 0.044) and GSH/GSSG ratios (p = 0.017) compared to controls. 129S1/SvImJ mice also exhibited lower hepatic GSH/GSSG ratios (p = 0.007) with C3G treatment. In this strain, C3G also increased GSSG levels in the liver (p = 0.02) as well as decreased total glutathione in the heart (p = 0.034) compared to controls.

Conclusions

To our knowledge, this study is the first to show that the biological effects of anthocyanins are determined by genotype. This study also demonstrates that these effects occur in a tissue‐specific manner. Our data will inform future efforts to clarify genetic mechanisms that regulate differential responses to anthocyanin supplementation.
Proteomic analysis of human serum of osteopenic women after dried plum treatment
Authors: Lee Fanny , Ghassemian Majid , Schenk Simon , Kern Mark , Hooshmand Shirin
Published: November 2023

Dried plum has the ability to prevent and reverse bone loss as evidenced by an increase in bone mineral density as well as changes in bone turnover biomarkers such as insulin growth factor‐1, bone‐specific alkaline phosphatase, tartrate‐resistant acid phosphatase‐5b and sclerostin. However, it is not known whether dried plum affects other proteins in human serum that may influence bone turnover. The objective of this study was to explore if dried plum affects proteins in human serum other than the known bone turnover biomarkers related in the reversal of bone loss in osteopenic postmenopausal women using isobaric tags for relative and absolute quantification (iTRAQ). Serum samples were obtained from forty‐eight osteopenic women (65–79 years old) who were randomly assigned to one of three treatment groups for six months: 1) control 2) 100g dried plum or 3) 50g dried plum. Samples were collected at baseline and after three and six months of intervention. The iTRAQ quantitative proteomics approach was conducted to explore the proteome affected by dried plum treatment. iTRAQ results identified 51 unique proteins that were upregulated with at least a 2‐fold difference in change between control and treatment groups (p<0.05). Twenty‐four proteins were significantly upregulated only at 3‐months, five proteins at both 3‐ and 6‐months and eighteen at 6‐months only. Of the relevant proteins, ten proteins relate to inflammation, at least 5 relate to immunological function in general, five relate to lipid metabolism and one has a known direct association with bone metabolism. Dried plum treatment resulted in an upregulation of 51 unique proteins in human serum, which may or may not be linked to bone turnover. Future studies must be conducted to determine if these are in fact related to bone turnover processes.

Support or Funding Information

Pilot Award from UCSD CTRI (UL1 TR000100)
Resveratrol and pterostilbene inhibit TNF‐α stimulated activation of NF‐kB and its upstream TAK1 in macrophages via modulation of sphingolipids
Authors: Yang Chao , Jiang Qing
Published: November 2023

Nuclear factor‐kB (NF‐kB) plays a key role in inflammatory responses and cancer development. Inhibition of NF‐kB has been shown to attenuate inflammation and cancer progression. Resveratrol (Res), a polyphenol rich in grapes and red wine, has been reported to inhibit NF‐kB signaling in macrophages. However, the underlying mechanism is not completely understood. Pterostilbene (Pte), a natural dimethylated analog of Res found in blueberries, is believed to have higher bioavailability than Res. While there are few studies on the effect of Pte on NF‐kB signaling, here we compared the effect of Res and Pte on modulating NF‐kB signaling and investigated potential underlying mechanisms in Raw 264.7 macrophages. Both Res and Pte inhibited TNF‐α induced phosphorylation of IkBα in a time‐ and dose‐ dependent manner. Res showed stronger inhibitory effect than Pte in this activity. Consistently, Res was stronger than Pte in blocking TNF‐α induced degradation of IkBα and inhibiting p65 phosphorylation in the nucleus. Furthermore, we found that both Res and Pte potently inhibited TNF‐α‐induced phosphorylation of TAK1, a key upstream signaling for NF‐kB activation. In search for the underlying mechanisms, we observed that Res treatment induced intracellular elevation of ceramides, sphingosine and dihydrosphingosine. Intriguingly, co‐treatment of C17 sphingosine and C8 ceramide attenuated TNF‐α‐induced phosphorylation of IkBα, mimicking the effect of Res. Our study demonstrates that Res inhibits NF‐kB and its upstream signaling TAK1 possibly via modulation of sphingolipid metabolism. Studies are undertaken to further investigate modulation effect of Pte on sphingolipid metabolism.
Naringenin, a citrus flavanone, enhances isoproterenol‐stimulated thermogenic activation of 3T3‐L1 adipocytes
Authors: Zhao Ling , Bae Jiyoung , Chen Jiangang
Published: November 2023

Promoting thermogenesis through increasing the activities of functional brown adipocytes has promised new hope for obesity treatment and prevention. Recent evidence has suggested that naringenin, a citrus flavanone, is beneficial for preventing obesity and obesity associated chronic diseases and naringenin is capable of activating nuclear receptors, including PPARγ; however, it has not been reported whether naringenin promotes activities of functional brown adipocytes, similar to what has been reported for rosiglitazone, a synthetic ligand for PPARγ. Here, we investigated the effects of naringenin on browning and isoproterenol (ISO)‐stimulated thermogenic activation of 3T3‐L1 adipocytes. 3T3‐L1 cells were induced to differentiate in the presence or absence of naringenin, followed by stimulation with ISO. We report that naringenin did not increase UCP‐1 expression at basal state; however, it significantly increased the ISO‐stimulated upregulation of UCP‐1 and PGC‐1α and enhanced ISO‐stimulated thermogenesis. Moreover, naringenin enhanced PKA activation and phosphorylation of p38 MAPK downstream of ISO stimulation; and inhibition of either PKA or p38 by their respective pharmacological inhibitors blocked naringenin‐induced upregulation of UCP‐1 mRNA. Our results demonstrate that naringenin enhances ISO‐induced thermogenic activation of 3T3‐L1 adipocytes via PKA/p38 pathways.

Support or Funding Information

The work was supported in part by UT professional development award to LZ.
Diosmetin: An Anti‐Leukemic Flavonoid Targeting the Estrogen Receptor β
Authors: Rota Sarah G. , Roma Alessia , Spagnuolo Paul A.
Published: November 2023

Acute myeloid leukemia (AML) is an aggressive hematological malignancy resulting from the accumulation of immature myeloid cells in the peripheral blood and bone marrow. It is associated with poor prognosis, especially in older patients where the 2‐year survival rate is approximately 10%. Leukemia stem cells (LSCs) are the cells responsible for AML development. To prevent relapse and improve patient prognosis, new therapies targeting LSCs are needed. Thus, to identify novel anti‐AML agents, we created a unique library consisting of food‐derived bioactive compounds (i.e., nutraceuticals). We screened this library (n= 288) using the MTS assay against TEX cells, a surrogate LSC line. Among the 288 compounds, diosmetin, a flavonoid derived from various citrus fruits, was found to be the most potent (EC50: 6.1 ± 0.5 μM, p<0.001). Most importantly, diosmetin did not affect the non‐LSC K562 cells, suggesting that diosmetin may selectively target leukemia stem cells. This was further demonstrated by clonogenic growth assays which showed that diosmetin (10 μM) reduced clonogenic growth in primary AML patient samples (n=6) but not in CD34+ normal hematopoietic cells (n=3). Finally, AML mouse xenografts showed that, compared to vehicle control, diosmetin treated mice had slower tumor growth and reduced tumor weights up on sacrifice (p<0.01). Collectively, this highlights diosmetin as a novel and selective anti‐AML agent.

Diosmetin induced caspase 3 mediated apoptosis, as determined by caspase activation, Annexin V/Propidium iodide and DNA fragmentation assays. To identify diosmetin's molecular target we utilized online bioinformatics tools (e.g., protein data base, PoSSuM and DAVID tool), which identified the estrogen receptor (ER) as diosmetin's potential molecular target. To assess the cell and molecular role of ERs, we measured ERα and ERβ levels in diosmetin sensitive and insensitive cell lines. Interestingly, diosmetin sensitive cell lines (TEX, LP1) display significantly elevated ERβ protein and mRNA levels (4 fold, as determined by Western blotting and qtPCR, respectively) compared to diosmetin insensitive cell lines (K562, DU145). This pattern was not observed for ERα. Furthermore, this ER expression pattern was also observed in patient‐derived AML cells, as cells sensitive to diosmetin displayed elevated ERβ, but not ERα, mRNA. Genetic knockdown using shRNA confirmed that ERβ is diosmetin's target, as cells lacking ERβ were resistant to diosmetin. Finally, ER reporter assays demonstrated that diosmetin binds and acts as an agonist in ERβ but not ERα reporter cells. Together, these results show that diosmetin binds to ERβ and that ERβ is functionally important to diosmetin's activity.

In summary, these studies highlight diosmetin binding to ERβ as a potential novel strategy for the treatment of AML.

Support or Funding Information

Stem Cell Network, University of Waterloo
TURMERIC POWDER GAVAGE IMPROVES LIPID PROFILE OF NORMAL ALBINO RATS
Authors: Iwueke Adaku Vivien , Iheanacho Kizito , Nwodu Judith
Published: November 2023

Dyslipidemia is one of the key risk factors for Cardiovascular disease, which is a leading cause of global morbidity and mortality. This study was designed to investigate the effect of powdered Curcuma longa (turmeric) tubers on lipid profile of normolipidemic albino rats. Twenty four male albino rats were randomly divided into four groups (A–D), receiving once daily administration of distilled water, 50, 100 and 200 mg/kg of the powder respectively. The body weights of the animals were measured weekly, while lipid profile was determined after three weeks. The result showed that daily oral administration of graded doses of the C. longa powder for three weeks resulted in a non‐significant increase in the body weights of the animals. Daily oral turmeric powder (200 mg/kg) treatment significantly (p ≤ 0.05) lowered the serum total cholesterol and triglyceride concentrations, while those that received 100 mg/kg significantly increased HDL and reduced LDL‐Cholesterol.

Turmeric powder has positive effect on lipid profile by lowering total cholesterol, triglyceride and LDL cholesterol and increasing HDL cholesterol in albino rats at the indicated doses.

Support or Funding Information

NIL