Pentagalloylglucose Research Articles

Ultrasound-microwave-assisted extraction of polyphenolic compounds from Mexican "Ataulfo" mango peels: Antioxidant potential and identification by HPLC/ESI/MS.

Mango is used in traditional medicine in many countries. However, the processing by-products are not currently used and generate large pollution problems and high handling costs. To study the effect of different parameters in the extraction of polyphenolic compounds from mango peels using modern and ecological ultrasound-microwave-assisted extraction technology. Various parameters of these processes were studied: the extract was recovered by liquid chromatography using Ambetlite XAD-16. The total polyphenol content was determined by Folin-Ciocalteu's and HCl-butanol methods. Antioxidant activity was determined by 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid (ABTS+), 1,10-diphenyl-2-20-picrylhydrazyl (DPPH), and lipid oxidation inhibition methods. The recovered compounds were identified by high-performance liquid chromatography-mass spectrometry (HPLC-MS). The best extraction conditions were solid/liquid ratio of 1/5 g/mL, ethanol percentage of 50%, and an extraction time of 10 min. Under these conditions, the total polyphenol content was 54.15 mg/g, and the antioxidant activities were greater than 90% inhibition in the three assays evaluated. According to the high-performance liquid chromatography/electrospray ionization/mass spectrometry (HPLC/ESI/MS) analysis, nine polyphenolic compounds were identified; most of them were gallotannins, such as pentagalloyl glucose. Ultrasound-microwave-assisted extraction was shown to be effective and allowed the recovery of antioxidant polyphenolic compounds. The results indicated that mango peel extracts can be used as natural antioxidant components in the pharmaceutical and functional food industries.

Pentagalloyl glucose from Schinus terebinthifolia inhibits growth of carbapenem-resistant Acinetobacter baumannii

The rise of antibiotic resistance has necessitated a search for new antimicrobials with potent activity against multidrug-resistant gram-negative pathogens, such as carbapenem-resistant Acinetobacter baumannii (CRAB). In this study, a library of botanical extracts generated from plants used to treat infections in traditional medicine was screened for growth inhibition of CRAB. A crude extract of Schinus terebinthifolia leaves exhibited 80% inhibition at 256 µg/mL and underwent bioassay-guided fractionation, leading to the isolation of pentagalloyl glucose (PGG), a bioactive gallotannin. PGG inhibited growth of both CRAB and susceptible A. baumannii (MIC 64–256 µg/mL), and also exhibited activity against Pseudomonas aeruginosa (MIC 16 µg/mL) and Staphylococcus aureus (MIC 64 µg/mL). A mammalian cytotoxicity assay with human keratinocytes (HaCaTs) yielded an IC50 for PGG of 256 µg/mL. Mechanistic experiments revealed iron chelation as a possible mode of action for PGG’s activity against CRAB. Passaging assays for resistance did not produce any resistant mutants over a period of 21 days. In conclusion, PGG exhibits antimicrobial activity against CRAB, but due to known pharmacological restrictions in delivery, translation as a therapeutic may be limited to topical applications such as wound rinses and dressings.

Intraluminal infusion of Penta-Galloyl Glucose reduces abdominal aortic aneurysm development in the elastase rat model.

An abdominal aortic aneurysm (AAA) is a progressive chronic dilatation of the abdominal aorta with terminally rupture when the aortic wall is so weakened that aortic wall stress exceeds wall strength. No effective medical treatment exists so far. We aimed to test whether intraluminal admission of Penta-Galloyl Glucose (PGG) treatment in a rodent AAA model could hold the potential to inhibit aneurysmal progression. Male Sprague Dawley rats had either intraluminal elastase infused for AAA induction or saline to serve as controls. In two independent experimental series, elastase was used to induce AAA followed by an intraluminal PGG (directly or by a drug eluting balloon) treatment. All rats were followed for 28 days and euthanized. In both series, maximal infrarenal aortic diameter was measured at baseline and at termination as a measure of AAA size. In series 2, maximal internally AAA diameter was followed by ultrasound weekly. AAA tissues were analyzed for elastin integrity by millers stain, collagen deposition by masson trichrome staining. In other AAA tissue samples the mRNA level of CD45, lysyloxidase (LOX), lysyloxidase like protein 1 (LOXL1) were determined by qPCR. Direct administration of PGG significantly reduced AAA expansion when compared to controls. PGG treatment resulted in a higher number and more preserved elastic fibers in the aneurysmal wall, while no significant difference was seen in the levels of CD45 and LOX mRNA levels. The drug eluting balloon (DEB) experiment showed no significant difference in AAA size observed neither macroscopically nor ultrasonically. Also the aneurysmal mRNA levels of CD45, LOX and LOXL1 were unchanged between groups. A significant reduced expansion of AAAs was observed in the PGG group, suggesting PGG as a drug to inhibit aneurysmal progression, while administration through a DEB did not show a promising new way of administration.

Pentagalloyl glucose (PGG) partially prevents arterial mechanical changes due to elastin degradation.

Elastic fibers are composed primarily of the protein elastin and they provide reversible elasticity to the large arteries. Degradation of elastic fibers is a common histopathology in aortic aneurysms. Pentagalloyl glucose (PGG) has been shown to bind elastin and stabilize elastic fibers in some in vitro studies and in vivo models of abdominal aortic aneurysms, however its effects on native arteries are not well described. Perform detailed studies of the biomechanical effects of PGG on native arteries and the preventative capabilities of PGG for elastin degraded arteries. We treated mouse carotid arteries with PGG, elastase (ELA), and PGG+ELA and compared the wall structure, solid mechanics, and fluid transport properties to untreated (UNT) arteries. We found that PGG alone decreased compliance compared to UNT arteries, but did not affect any other structural or biomechanical measures. Mild (30 sec) ELA treatment caused collapse and fragmentation of the elastic lamellae, plastic deformation, decreased compliance, increased modulus, and increased hydraulic conductance of the arterial wall compared to UNT. PGG+ELA treatment partially protected from all of these changes, in particular the plastic deformation. PGG mechanical protection varied considerably across PGG+ELA samples and appeared to correlate with the structural changes. Our results provide important considerations for the effects of PGG on native arteries and a baseline for further biomechanical studies on preventative elastic fiber stabilization.

Structural characteristics of pharmacodynamical components in genuine Moutan Cortex

Guided by the theory of "component structure", we analyzed the structural characteristics of pharmacodynamical components in genuine Moutan Cortex. The compositions of organic small molecules were determined by high performance liquid chromatography(HPLC) for 20 batches of genuine Moutan Cortex and 12 batches of non-genuine Moutan Cortex. By means of similarity analysis, clustering analysis(CA), principal component analysis(PCA) and orthogonal partial least-squares discriminant analysis(OPLS-DA), the elements in structural characteristics of the pharmacodynamical components were extracted as follows: terpene glycosides components(oxidized paeoniflorin, paeoniflorin,galloyl paeoniflorin, benzoyloxy paeoniflorinand benzoyl paeoniflorin), tannin components(1,3,6-tri-O-galloside acyl glucose, pentagalloyl glucose), and phenolic acid components(methyl gallate, paeonol). The contents and quantity ratios of terpene glycoside component, tannin component and phenolic acid components in genuine Moutan Cortex were determined as 14.1, 12.5, 21.7 mg·g~(-1), 1.00∶0.89∶1.54. The contents and quantity ratios of the oxidized paeoniflorin, paeoniflorin and benzoylpaeoniflorin in the terpene glycoside components were characteristic and determined as 2.05, 7.05, 3.30 mg·g~(-1), 1.00∶3.44∶1.61. The unique structural characteristics of genuine Moutan Cortex provide scientific basis for the formulation of quality standards.

Characterization and Anti-stroke Activity of Stem Extract of Colebrookea oppositifolia Smith

This study was aimed to investigate the cerebroprotective potential of methanolic stem extract of Colebrookea oppositifolia (MsCO). The MsCO was evaluated against global ischemia and reperfusion (I/R) induced brain injury in Wistar rats. In the outcomes, the I/R insult caused significant deterioration of neurological (p<0.01), cognitive (p<0.01), and motor functions (p<0.01), along with significant alterations in the antioxidant parameters (decreased SOD and catalase enzyme activities, along with derease in GSH and increase in LPO; p<0.01 for all parameters), morphology (cerebral edema and infarct area) (p<0.01) and histology of the brain tissue in the I/R control group compared to normal control. Interestingly, the MsCO (200 and 400 mg/kg) pretreatment for seven days has attenuated all the I/R-induced alterations significantly and dose-dependently compared to I/R control group. Moreover, the LC-MS/MS analysis of MsCO revealed the presence of Acteoside, Pentagalloyl glucose, and Quercetin as major phytoconstituents. Based on these outcomes we conclude that MsCO possesses potent cerebroprotective activity against ischemia-reperfusion-induced brain injury in Wistar rats.

Studies on interactions of pentagalloyl glucose, ellagic acid and gallic acid with bovine serum albumin: A spectroscopic analysis

Fluorescence techniques and circular dichroism (CD) were utilized to investigate the interactions of a typical gallotannin 1,2,3,4,6-penta-O-galloyl-β-D-glucopyranose (PGG) and two simple phenolic compounds ellagic acid (EA) and gallic acid (GA) with bovine serum albumin (BSA). Fluorescence experiments showed that PGG and EA could strongly interact with BSA. The binding constants showed a pH-dependent binding of phenolic acids by BSA. The CD spectra revealed that GA, EA and PGG has slight effect on the secondary structure of BSA at the low concentration (50 μM). However, obvious alterations of the secondary structure of BSA were observed at high concentrations (from 100 μM to 500 μM). The interaction of GA, EA and PGG at high concentrations with BSA caused an unfolding of the secondary structure of BSA. PGG has a greater impact on the secondary structure of BSA when compared with small molecule compounds GA and EA.

The role of NFκB in pentagalloyl glucose inhibition of TNF‐α‐induced CXCL1 expression and induction of apoptosis‐related genes in racially different triple negative breast cancer cells.

Drug resistance is the leading cause of breast cancer‐related mortality in women, and triple negative breast cancer (TNBC) is the most aggressive subtype. CXCL1/GRO‐α has been identified as a pro‐inflammatory chemokine overexpressed in breast cancer, promoting angiogenesis and having an increased potential to metastasize. The current study investigated the effect of the naturally occurring polyphenolic compound pentagalloyl glucose (PGG) in TNF‐α‐induced CXCL1 expression in MDA‐MB‐231 (Caucasian) and MDA‐MB‐468 (African American) TNBC cells. In addition, PGG apoptotic effect was evaluated through the study of apoptosis‐related genes in both cell lines. Results showed that PGG inhibited TNF‐α‐stimulated CXCL1 expression in the transcription and protein level in Caucasian and African American TNBC cell lines. Through mRNA quantification analysis, data showed that PGG downregulated IKBKE (expressed in 60.4% of the breast cancer tumors) and MAPK1/ERK2 (expressed in tumor tissues with 5 to 10‐fold increase) expression in both cell lines. Moreover, PGG was more effective in inhibiting IKBKE expression in MDA‐MB‐231 compared to MDA‐MB‐468 (9 and 2‐fold inhibition, respectively). Also, PGG reduced MAPK1 gene expression in 3‐fold in Caucasian and 6‐fold in the African American cells. Gene transcription studies showed that PGG induced expression of the pro‐apoptotic genes BNIP3, BNIP3L, GADD45A, TP53BP2, and RIPK2, which are critical initiators of the intrinsic and extrinsic apoptotic pathways. Although PGG increased the expression of apoptosis‐related genes in both cell lines, there was a significantly higher expression in MDA‐MB‐231 cells (5 to 14.7‐fold) compared to MDA‐MB‐468 cells (2 to 5.8‐fold). Furthermore, BAK1 gene expression was only induced in African American cells showing a 35.7‐fold increase after PGG treatment. The obtained data show that PGG inhibited the expression of CXCL1 pro‐inflammatory chemokine and upregulated the expression of specific pro‐apoptotic genes. It was concluded that PGG might have great potential in target therapy for TNBC.Support or Funding InformationThis work was supported by a grant from the NIH/NIMHD grant U54 007582

Pentagalloyl glucose (PGG) Inhibits IL‐1β‐induced pro‐inflammatory cytokine production and matrix metalloproteinase activation in human osteoarthritis synovial fibroblasts

Osteoarthritis (OA) is a chronic degenerative joint disease in which the pro‐inflammatory cytokine, interleukin‐1β (IL‐1β), plays a central role in propagating inflammation and cartilage degradation by activating inflammatory mediators and cartilage degrading matrix metalloproteinases (MMPs). In the present study, we evaluated the efficacy of pentagalloyl glucose (PGG), a natural antioxidant, in suppressing IL‐1β‐induced mediators of inflammation and cartilage degradation in human osteoarthritis synovial fibroblasts (OASFs). PGG treatment (0.5–50 μM) for 24 hours had no significant effect on human OASF cell viability (n=3; p&lt;0.01). Analysis of the conditioned media using ELISA showed that PGG (0.5–5 uM/ml) pretreatment significantly inhibited IL‐1β‐induced production of IL‐6 (45–90%), IL‐8 (40–95%), MCP‐1/CCL2 up to 80%, and RANTES/CCL5 by (45–90%) (n=3; p&lt;0.05). In addition, we observed a significant dose‐dependent inhibition in MMP‐1 and MMP‐3 production by PGG in human OASFs (n=3; p&lt;0.05). Furthermore, in‐gel zymography results using conditioned media showed that the enzymatic activity of IL‐1β‐induced gelatinases MMP‐2 and MMP‐9 were markedly reduced upon PGG treatment (n=3; p&lt;0.05). PGG (5 μg/mL) significantly inhibited the degradation of type‐I collagen compared to IL‐1β stimulation alone in human OASFs (n=3; p&lt;0.05). Examination of the mitogen‐activated protein kinase (MAPK) signaling pathway showed that PGG interfered with IL‐1β‐induced ERK1/2 and JNK/SAPK, but not p38 MAPK, to inhibit downstream phosphorylation and nuclear translocation of transcription factor c‐Jun. Immunofluorescence study confirmed that PGG pretreatment inhibits phospho‐c‐Jun and NF‐κB p65 nuclear translocation. Overall, our findings suggest that PGG inhibits IL‐1β‐induced mediators of inflammation and tissue destruction in human OASFs by reducing the activation of MAPK signaling pathway.Support or Funding InformationThis study was supported by the NIH/NIAMS grant R01AR072615 (S.A.), and the Washington State University College of Pharmacy (S.A.), the Summer Undergraduate Research Fellowship from ASPET (M.T.), and Special thanks to Erwin &amp; Jennie Foisy Scholarship Fund (M.H.).

Nanoparticle-based targeted delivery of pentagalloyl glucose reverses elastase-induced abdominal aortic aneurysm and restores aorta to the healthy state in mice.

AimAbdominal aortic aneurysms (AAA) is a life-threatening weakening and expansion of the abdominal aorta due to inflammatory cell infiltration and gradual degeneration of extracellular matrix (ECM). There are no pharmacological therapies to treat AAA. We tested the hypothesis that nanoparticle (NP) therapy that targets degraded elastin and delivers anti-inflammatory, anti-oxidative, and ECM stabilizing agent, pentagalloyl glucose (PGG) will reverse advance stage aneurysm in an elastase-induced mouse model of AAA.Method and resultsPorcine pancreatic elastase (PPE) was applied periadventitially to the infrarenal aorta in mice and AAA was allowed to develop for 14 days. Nanoparticles loaded with PGG (EL-PGG-NPs) were then delivered via IV route at 14-day and 21-day (10 mg/kg of body weight). A control group of mice received no therapy. The targeting of NPs to the AAA site was confirmed with fluorescent dye marked NPs and gold NPs. Animals were sacrificed at 28-d. We found that targeted PGG therapy reversed the AAA by decreasing matrix metalloproteinases MMP-9 and MMP-2, and the infiltration of macrophages in the medial layer. The increase in diameter of the aorta was reversed to healthy controls. Moreover, PGG treatment restored degraded elastic lamina and increased the circumferential strain of aneurysmal aorta to the healthy levels.ConclusionOur results support that site-specific delivery of PGG with targeted nanoparticles can be used to treat already developed AAA. Such therapy can reverse inflammatory markers and restore arterial homeostasis.

Enhancement of Biomimetic Enzymatic Mineralization of Gellan Gum Polysaccharide Hydrogels by Plant-Derived Gallotannins.

Mineralization of hydrogel biomaterials with calcium phosphate (CaP) is considered advantageous for bone regeneration. Mineralization can be both induced by the enzyme alkaline phosphatase (ALP) and promoted by calcium-binding biomolecules, such as plant-derived polyphenols. In this study, ALP-loaded gellan gum (GG) hydrogels were enriched with gallotannins, a subclass of polyphenols. Five preparations were compared, namely three tannic acids of differing molecular weight (MW), pentagalloyl glucose (PGG), and a gallotannin-rich extract from mango kernel (Mangifera indica L.). Certain gallotannin preparations promoted mineralization to a greater degree than others. The various gallotannin preparations bound differently to ALP and influenced the size of aggregates of ALP, which may be related to ability to promote mineralization. Human osteoblast-like Saos-2 cells grew in eluate from mineralized hydrogels. Gallotannin incorporation impeded cell growth on hydrogels and did not impart antibacterial activity. In conclusion, gallotannin incorporation aided mineralization but reduced cytocompatibility.

Anti-influenza virus phytochemicals from Radix Paeoniae Alba and characterization of their neuraminidase inhibitory activities

Ethnopharmacological relevanceBai Shao (Radix Paeoniae Alba, BS), the root of Paeonia lactiflora Pall., in ancient China was used for Wen Bing (Warm Disease) treatment. Wen Bing has the symptoms of influenza. Ethanol extract of the root has recently been shown to possess anti-influenza activity. However, the active compounds have not yet been identified. AimWe showed that BS aqueous extract was potent in inhibiting influenza A virus in infected cells. We aimed to isolate the bioactive compounds and characterize the anti-influenza mechanism. Materials and methodsPlaque reduction assay was performed for fractions isolated from BS. Hemagglutination inhibition assay and neuraminidase inhibition assay were performed to find the target protein. Molecular docking and reverse genetics were used to confirm the action site of gallic acid on the neuraminidase protein. ResultsWe identified three tannin compounds gallic acid (GA), methyl gallate (MG) and pentagalloylglucose (PGG) in BS aqueous extract that could inhibit the replication of influenza A virus in MDCK cells. While only PGG was found to inhibit the influenza virus-induced hemagglutination of chicken erythrocytes, all three compounds significantly reduced the activity of the neuraminidase. The results from molecular docking and reverse genetics showed that GA interacted with Arg152 of neuraminidase protein. ConclusionThree compounds GA, MG and PGG isolated from BS were found to inhibit influenza A virus in MDCK cells. GA interacts with amino acid Arg152 of the viral neuraminidase. Our study identified anti-influenza compounds of BS and demonstrated their antiviral mechanism, thus providing scientific evidence for using this herb for clinical treatment.

Pentagalloyl Glucose- and Ethyl Gallate-Rich Extract from Maprang Seeds Induce Apoptosis in MCF-7 Breast Cancer Cells through Mitochondria-Mediated Pathway.

Bouea macrophylla Griffith, locally known as maprang, has important economic value as a Thai fruit tree. The maprang seed extract (MPSE) has been shown to exhibit antibacterial and anticancer activities. However, the bioactive constituents in MPSE and the molecular mechanisms underlying these anticancer activities remain poorly understood. This study aims to identify the active compounds in MPSE and to investigate the mechanisms involved in MPSE-induced apoptosis in MCF-7 treated cancer cells. The cytotoxic effect was determined by MTT assay. The apoptosis induction of MPSE was evaluated in terms of ROS production, mitochondrial membrane potential depolarization, and apoptosis-related gene expression. The compounds identified by HPLC and LC/MS analysis were pentagalloyl glucose, ethyl gallate, and gallic acid. MPSE treatment decreased cell proliferation in MCF-7 cells, and MPSE was postulated to induce G2/M phase cell cycle arrest. MPSE was found to promote intracellular ROS production in MCF-7 treated cells and to also influence the depolarization of mitochondrial membrane potential. In addition, MPSE treatment can lead to increase in the Bax/Bcl-2 gene expression ratio, suggesting that MPSE-induced apoptosis is mitochondria-dependent pathway. Our results suggest that natural products obtained from maprang seeds have the potential to target the apoptosis pathway in breast cancer treatments.

Mechanism of Pentagalloyl Glucose in Alleviating Fat Accumulation in Caenorhabditis elegans.

Pentagalloyl glucose (PGG) has been studied for its valuable biological activities. However, the functional role of PGG in lipid metabolism in vivo is unclear. Here, we investigated the effects of PGG on lipid metabolism and its underlying mechanism in Caenorhabditis elegans. PGG decreased the accumulation of reactive oxygen species at 800 μM and remarkably increased the activities of antioxidant enzymes. PGG decreased significantly fat accumulation in wild-type worms (39.7 ± 5.7% in the normal group and 19.9 ± 4.5% in the high-fat group by Oil red O; 21.2 ± 2.7% in the high-fat group by Nile red; p < 0.001), but fat reduction by PGG was eliminated in the skn-1 mutant. The amount and size of lipid droplets in the ZXW618 mutant were decreased by PGG. The proportions of unsaturated fatty acids in both conditions were increased by PGG. In addition, the expression levels of fat metabolism genes were significantly changed in both conditions by PGG, which include mdt-15, pod-2, elo-2, fat-6, and fat-7 genes modulated fat synthesis; aak-2 and nhr-49 genes participated in fat consumption; and tub-1 gene regulated fat storage. However, fat-5 and acs-2 were downregulated in high-fat worms only, and vit-2 and lipl-4 were downregulated in normal worms only. Our study provided new insights into the role of PGG in alleviating fat accumulation and its underlying mechanism of action in C. elegans.

Pentagalloylglucose Inhibits the Replication of Rabies Virus via Mediation of the miR-455/SOCS3/STAT3/IL-6 Pathway.

Our previous study showed that pentagalloylglucose (PGG), a naturally occurring hydrolyzable phenolic tannin, possesses significant anti-rabies virus (RABV) activity. In BHK-21 cells, RABV induced the overactivation of signal transducer and activator of transcription 3 (STAT3) by suppressing the expression of suppressor of cytokine signaling 3 (SOCS3). Inhibition of STAT3 by niclosamide, small interfering RNA, or exogenous expression of SOCS3 all significantly suppressed the replication of RABV. Additionally, RABV-induced upregulation of microRNA 455-5p (miR-455-5p) downregulated SOCS3 by directly binding to the 3' untranslated region (UTR) of SOCS3. Importantly, PGG effectively reversed the expression of miR-455-5p and its following SOCS3/STAT3 signaling pathway. Finally, activated STAT3 elicited the expression of interleukin-6 (IL-6), thereby contributing to RABV-associated encephalomyelitis; however, PGG restored the level of IL-6 in vitro and in vivo in a SOCS3/STAT3-dependent manner. Altogether, these data identify a new miR-455-5p/SOCS3/STAT3 signaling pathway that contributes to viral replication and IL-6 production in RABV-infected cells, with PGG exerting its antiviral effect by inhibiting the production of miR-455-5p and the activation of STAT3.IMPORTANCE Rabies virus causes lethal encephalitis in mammals and poses a serious public health threat in many parts of the world. Numerous strategies have been explored to combat rabies; however, their efficacy has always been unsatisfactory. We previously reported a new drug, PGG, which possesses a potent inhibitory activity on RABV replication. Herein, we describe the underlying mechanisms by which PGG exerts its anti-RABV activity. Our results show that RABV induces overactivation of STAT3 in BHK-21 cells, which facilitates viral replication. Importantly, PGG effectively inhibits the activity of STAT3 by disrupting the expression of miR-455-5p and increases the level of SOCS3 by directly targeting the 3' UTR of SOCS3. Furthermore, the downregulated STAT3 inhibits the production of IL-6, thereby contributing to a reduction in the inflammatory response in vivo Our study indicates that PGG effectively inhibits the replication of RABV by the miR-455-5p/SOCS3/STAT3/IL-6-dependent pathway.

Biomechanical Restoration Potential of Pentagalloyl Glucose after Arterial Extracellular Matrix Degeneration.

The objective of this study was to quantify pentagalloyl glucose (PGG) mediated biomechanical restoration of degenerated extracellular matrix (ECM). Planar biaxial tensile testing was performed for native (N), enzyme-treated (collagenase and elastase) (E), and PGG (P) treated porcine abdominal aorta specimens (n = 6 per group). An Ogden material model was fitted to the stress–strain data and finite element computational analyses of simulated native aorta and aneurysmal abdominal aorta were performed. The maximum tensile stress of the N group was higher than that in both E and P groups for both circumferential (43.78 ± 14.18 kPa vs. 10.03 ± 2.68 kPa vs. 13.85 ± 3.02 kPa; p = 0.0226) and longitudinal directions (33.89 ± 8.98 kPa vs. 9.04 ± 2.68 kPa vs. 14.69 ± 5.88 kPa; p = 0.0441). Tensile moduli in the circumferential direction was found to be in descending order as N > P > E (195.6 ± 58.72 kPa > 81.8 ± 22.76 kPa > 46.51 ± 15.04 kPa; p = 0.0314), whereas no significant differences were found in the longitudinal direction (p = 0.1607). PGG binds to the hydrophobic core of arterial tissues and the crosslinking of ECM fibers is one of the possible explanations for the recovery of biomechanical properties observed in this study. PGG is a beneficial polyphenol that can be potentially translated to clinical practice for preventing rupture of the aneurysmal arterial wall.

Proteomic analysis of the effect of the polyphenol pentagalloyl glucose on proteins involved in neurodegenerative diseases in activated BV‑2 microglial cells.

Neuroinflammation and microglial activation are two important hallmarks of neurodegenerative diseases. Continuous microglial activation may cause the release of several cytotoxic molecules, including many cytokines that are involved in the inflammatory process. Therefore, attenuating inflammation caused by activated microglia may be an approach for the therapeutic management of neurodegenerative diseases. In addition, many studies have reported that polyphenol pentagalloyl glucose (1,2,3,4,6-penta-O-galloyl-β-D-glucose; PGG) is a molecule with potent anti-inflammatory effects, such as inhibiting the release of proinflammatory cytokines. Our previous studies revealed that PGG attenuated the expression of two inflammatory cytokines (murine monocyte chemoattractant protein-5 and pro-metalloproteinase-9) in lipopolysaccharide/interferon γ-activated BV-٢ microglial cells. Additionally, PGG modulated the NF-κB and MAPK signaling pathways by altering genes and proteins, which may affect the MAPK cascade and NF-κB activation. The aim of the present study was to investigate the ability of PGG to modulate the expression of proteins released in activated BV-2 microglial cells, which may be involved in the pathological process of inflammation and neurodegeneration. Proteomic analysis of activated BV-2 cells identified 17 proteins whose expression levels were significantly downregulated by PGG, including septin-7, ataxin-2, and adenylosuccinate synthetase isozyme 2 (ADSS). These proteins were previously described as being highly expressed in neurodegenerative diseases and/or involved in the signaling pathways associated with the formation and growth of neuronal connections and the control of Alzheimer's disease pathogenesis. The inhibitory effect of PGG on ataxin-2, septin-7 and ADSS was further confirmed at the protein and transcriptional levels. Therefore, the obtained results suggest that PGG, with its potent inhibitory effects on ataxin-2, septin-7 and ADSS, may have potential use in the therapeutic management of neurodegenerative diseases.

Identifying Mitochondrial Targets for Pancreatic Cancer Employing Unconventional Combinations of Middle Eastern Phytoextracts with Raloxifene

PCa has a high mortality, substantial chemoresistance, and a poor 5‐year survival rate of 6 to 8%. It is the fourth leading cause of deaths due to cancer in the United States, killing over 53,000 Americans each year, and 250,000 people worldwide. Little is known about the regulation of mitophagy in PCa by drugs and dietary components. There is also a gap in knowledge concerning molecular interactions between drugs and food and the K‐ras pathway in PCa. This is important because mutations in the human homologue of the Kirsten Rat Sarcoma (KRAS) viral oncogene occur in 90% of PCa patients and drive the early development of PCa. Our previous work showed that enhanced mitophagy, which prevents the accumulation of damaged mitochondria in PCa cells was alleviated by repurposing the FDA‐approved selective estrogen receptor modulator raloxifene to treat PCa cells. Here, we used a combination of Middle Eastern dietary components (MEDC) and raloxifene to mitigate mitochondrial renewal and identify novel mitochondrial targets for PCa. We screened crude aqueous extracts from six spices and plants used in Middle Eastern and Indian cuisines, including artichoke (Cynara scolymus), curry leaf (Murraya koenigii), dill (Anethum graveolens), noni (Morinda citrifolia), olive (Olea europaea), rose (Rosa damascena), and sumac (Rhus coriaria) for cytotoxicity in the PCa cell lines, MIA PaCa‐2 (similar KRAS gene mutation as majority of patients) and PANC‐1 (less common KRAS gene mutation). Specifically, we treated PCa cells in vitro with exponentially increasing doses of aqueous extracts at 300 ng/mL, 3 μg/mL, 30 μg/mL, and 300 μg/mL for 48 hours followed by an MTT assays. Aqueous extracts of sumac and artichoke (AESA) induced maximum cytotoxicity, which was exacerbated by raloxifene (AESA‐R). To address whether cytotoxicity was limited to PCa cells, we used NIH‐3T3 cells as a non‐cancer control, showing a sparing effect. We also tested 1:1 combinations of AESA and found evidence of activity. To identify specific compounds, using a fractional approach based on preparative HPLC, 100 mg of the residue was chromatographed onto a silica gel column and gradient eluted using dichloromethane/methanol (gradient, 0, 100%, v/v). Fractions and eluates were monitored by thin layer chromatography using precoated silica gel 60 F254, 0.25 mm aluminum backed plates. Cytotoxic fractions were reconstituted in deuterated solvents and analyzed by NMR spectroscopy to identify pentagalloyl glucose as the bioactive component in sumac. To elucidate the mechanism for the cytotoxicity, we showed that AESA and AESA‐R activated caspase‐3 and 7, induced mitochondrial ROS, and reduced mitochondrial membrane potential in PCa cells, starting within 20 minutes and sustained for 48 hours following treatment. Western blot showed changes in the expression of proapoptotic (bim) and the translocation of antiapoptotic proteins (Bcl2), sustained over 48 hours. Our data demonstrate for the first time that specific food‐drug combinations alter mitochondrial biology and response including the regulation of mitophagy in PCa cells.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Pentagalloyl glucose effects on murine abdominal aortic aneurysms

An abdominal aortic aneurysm (AAA) is a dilation of the abdominal segment of the largest artery in the body and can be accompanied by significant risk of rupture and mortality. The current treatment, surgical repair, carries risks and complications. As such, there is need for less invasive therapies capable of curbing aneurysm growth. Here we use an AAA mouse model to evaluate the potential of pentagalloyl glucose (PGG) to suppress aneurysm growth. To induce aneurysms, 5.0 mg/mL pancreatic porcine elastase (PPE) was topically applied to the infrarenal aorta and 0.2% beta aminoproprionitrile was continuously administered via drinking water. Four of the eight animals also received topical treatment of 0.06% PGG via gauze before PPE treatment. High frequency ultrasound imaging (Vevo2100 system, VisualSonics) with a MS550D transducer (40 MHz center frequency) was performed prior to surgery, and every week thereafter for 4 weeks. PGG-treated animals had a small but nonsignificant decrease (p = 0.09) in effective maximum aortic diameter on days 7 and 14 that was not present on day 28. Current work is being performed to quantify PGG binding via histological characterization. Ultimately, we aim to investigate the parameters required for PGG to be an effective treatment for mechanically-stabilizing small AAAs.

Methanolic leaf extract of Punica granatum attenuates ischemia-reperfusion brain injury in Wistar rats: Potential antioxidant and anti-inflammatory mechanisms.

Objective(s):This study was conducted to evaluate the cerebroprotective effect of methanolic leaf extract of Punica granatum (MePG) in Wistar rats.Materials and Methods:The MePG was initially assessed for in vitro antioxidant activity, and later evaluated on LPS-induced RAW 264.7 cell line assay. Finally, the MePG was evaluated against ischemia-reperfusion (I/R) induced brain injury in Wistar rats.Results:In DPPH, FRAP and ORAC assays, the MePG has exhibited potent antioxidant activity. Further, the MePG has significantly inhibited the generation of nitrite, ROS and TNF-α in LPS-induced RAW 264.7 cell lines. Besides, global ischemia followed by reperfusion caused significant changes in the neurological and behavioral functions in I/R control animals compared to sham control. Additionally, in the I/R control group there was a substantial decrease in the catalase and superoxide dismutase activities; Likewise, reduced glutathione levels reduced and lipid peroxidation levels enhanced significantly. Also, pro-inflammatory cytokines such as TNF-α, IL-6, and ICAM-I were increased and the levels of IL-10 was decreased significantly. Furthermore, the I/R insult caused increase in brain volume and cerebral infarct formation. Similarly, histopathology of the brain tissue revealed hallmarks like necrosis, leukocyte infiltration, cerebral edema and vascular congestion in I/R control. Notably, MePG (200 and 400 mg/kg) pretreatment for 7 days, has attenuated all the I/R-persuaded pathological changes compared to I/R control. In addition, the LC-MS/MS analysis showed presence of acteoside, apigenin, gallic acid, gossypin, pentagalloyl glucose, quercetin, and rutin as major ingredients in the MePG.Conclusion:These findings suggest that the MePG possesses significant cerebroprotective activity.