Different Concentrations Of Epigallocatechin-3-gallate Research Articles

Barnacle-inspired and polyphenol-assisted modification of bacterial cellulose-based wound dressings for promoting infectious wound healing

Bacterial cellulose (BC) is an ideal candidate for wound dressings due to its natural origin, exceptional water-holding capacity, pliability, biocompatibility, and high absorption capability. However, the lack of essential antimicrobial activity limits its biomedical applications. This study reported BC-based wound dressings containing silk fibroin protein (SF), offering the potential for biomimetic properties, and (−)-epigallocatechin-3-gallate (EGCG) for polyphenol-assisted surface modification to promote infectious wound healing. Glycerol was used as the carbon source to promote the formation of an adhesive layer by facilitating the β-sheet folding of SF, and different concentrations of EGCG were employed to interact with SF through strong hydrogen bonding facilitated by the polyphenolic groups. The functionalized membrane exhibited outstanding water-holding capacity, swelling ratio, and degradation properties, along with enhanced hydrophilicity, adhesiveness, and a remarkable free radical scavenging ability. Both in vitro and in vivo experiments confirmed its potent bacteriostatic activity. The composite membrane displayed excellent biocompatibility, including cellular and hemocompatibility. Importantly, it effectively promoted wound healing in murine back infections. These findings suggest the significant feasibility of the innovative modification approach, and that functionalized membranes have great potential as wound-dressing materials for infection management in future clinical applications.

P-631 Alleviating effects of EGCG on ovarian hyperstimulation syndrome and the potential modulating mechanism through the VEGF pathway

Abstract Study question What are the underlying mechanisms of EGCG to alleviate Ovary hyperstimulation syndrome (OHSS)? Summary answer EGCG inhibited VEGF via TGF-β1 classical-SMAD pathway and 67LR-mediated CREB pathway and could reduce the ovarian inflammatory effect and attenuated OHSS progress in rat model. What is known already Ovarian hyperstimulation syndrome (OHSS) is one of the most severe complications of COH during IVF treatment. The pathophysiology of OHSS is characterized by increased capillary permeability, VEGF is an important mediator in OHSS, and serum VEGF levels have been shown to correlate with OHSS severity. (-)-epigallocatechin-3-gallate (EGCG) is the most abundant and biologically active polyphenolic catechin in green tea and has been reported to have multiple effects in humans. Many research indicated that in many pathological processes, EGCG could inhibit VEGF and its receptor expression and have an angiogenesis effect. Herein, EGCG might have a therapeutic effect on OHSS. Study design, size, duration We investigated the role of EGCG in OHSS in vitro and in vivo. The primary human granulosa-lutein(hGL) cells and human granulosa-like tumor (KGN) cell line were cultured and treated with different concentrations of EGCG for 24 hours. Animal OHSS model was established in SD rats by injection of pregnant mare serum gonadotropin (PMSG), and randomly assigned to receive vehicle only or EGCG for 3 days. All experiments were performed 3-8 times for comparisons. Participants/materials, setting, methods The effect of EGCG on KGN and hGL cells was determined by MTT assay. The body weight of rats was measured every day, and the ovary size was measured after removal, the permeability was determined by Evans-blue. The serum estrogen and VEGF level in rats were detected by ELISA. RNA and protein expressions of VEGF, VEGFR-2, TGF-β, and TβR II were detected by qPCR and Western-blotting and immunostaining in vitro and in vivo experiments. Main results and the role of chance Our study demonstrated that administration of EGCG attenuated the development of OHSS in rats, as shown by histological examination and ovarian weight and morphology. The ovary weight was significantly decreased in the EGCG treatment group compared with the OHSS group (147.9 vs 206.5 mg). Additionally, compared to OHSS rats, EGCG treated rats exhibit downregulated ovarian VEGF expression determined by IHC and RT-qPCR. VEGF and E2 protein levels significantly decrease in the serum of the EGCG treatment group. EGCG exerted inhibitory effects on cell growth only in high dose (50uM) and longtime (48h) treatment in KGN and hGL cells. In KGN cells and hGL cells, EGCG significantly reduces the expression of VEGF and TGF-β at the RNA and protein levels. Furthermore, EGCG inhibits TGF-β1-induced VEGF production and secretion in KGN cells by suppressing TGF-β expression and its traditional Smad signaling pathway. EGCG also downregulates VEGF expression through the 67-kDa laminin receptor-mediated PKA-CREB pathway. Limitations, reasons for caution EGCG has been reported to employ a wide range of biological effects. Therefore, its suppressive effects on VEGF and TGF-β signaling pathway might be part of the pharmacological mechanisms to alleviate Ovary hyperstimulation syndrome. Further studies are also needed to explore the therapeutic mechanisms of melatonin from other perspectives. Wider implications of the findings Our findings add mechanistic insight in support of using EGCG as an adjuvant therapy in the management of OHSS. EGCG shows the potential to act as a novel alternative therapeutic drug to treat OHSS. Daily intake of green tea might be beneficial for women under COH to prevent OHSS. Trial registration number Not applicable

Epigallocatechin-3-gallate protects porcine oocytes against post-ovulatory aging through inhibition of oxidative stress.

Increased levels of oxidative stress are major factors that drive the process of post-ovulatory oocyte aging. Epigallocatechin-3-gallate (EGCG), which accounts for up to 50% of the catechins, possesses versatile biological functions, including preventing or treating diabetes, cancer, and heart diseases. The aim of this study was to explore whether EGCG can delay porcine oocyte aging by preventing oxidative stress. Metaphase II (MII) oocytes were cultured for 48 h with different concentrations of EGCG (0-100 μM) in vitro as a post-ovulatory aging model. An optimal concentration of 5 μM EGCG maintained oocyte morphology and developmental competence during aging. The oocytes were randomly divided into five groups: fresh, 24 h control, 24 h EGCG, 48 h control, and 48 h EGCG. The results suggest that EGCG significantly prevents aging-induced oxidative stress, glutathione (GSH) reduction, apoptosis, and autophagy. Moreover, mitochondria DNA copy number was decreased, and the number of active mitochondria and adenosine triphosphate (ATP) levels significantly increased by supplementation with EGCG. Thus, EGCG has a preventive role against aging in porcine post-ovulatory oocytes due to its ability to inhibit oxidative stress and promote mitochondrial biogenesis.

The combination of EGCG with warfarin reduces deep vein thrombosis in rabbits through modulating HIF-1α and VEGF via the PI3K/AKT and ERK1/2 signaling pathways.

Deep venous thrombosis (DVT) poses a major challenge to public health worldwide. Endothelial cell injury evokes inflammatory and oxidative responses that contribute to thrombus formation. Tea polyphenol (TP) in the form of epigallocatechin-3-gallate (EGCG) has anti-inflammatory and oxidative effect that may ameliorate DVT. However, the precise mechanism remains incompletely understood. The current study was designed to investigate the anti-DVT mechanism of EGCG in combination with warfarin (an oral anticoagulant). Rabbits were randomly divided into five groups. A DVT model of rats was established through ligation of the inferior vena cava (IVC) and left common iliac vein, and the animals were orally administered with EGCG, warfarin, or vehicle for seven days. In vitro studies included pretreatment of human umbilical vein endothelial cells (HUVECs) with different concentrations of EGCG for 2 h before exposure to hydrogen peroxide. Thrombus weight and length were examined. Histopathological changes were observed by hematoxylin-eosin staining. Blood samples were collected for detecting coagulation function, including thrombin and prothrombin times, activated partial thromboplastin time, and fibrinogen levels. Protein expression in thrombosed IVCs and HUVECs was evaluated by Western blot, immunohistochemical analysis, and/or immunofluorescence staining. RT-qPCR was used to determine the levels of AGTR-1 and VEGF mRNA in IVCs and HUVECs. The viability of HUVECs was examined by CCK-8 assay. Flow cytometry was performed to detect cell apoptosis and ROS generation was assessed by 2',7'-dichlorofluorescein diacetate reagent. In vitro and invivo studies showed that EGCG combined with warfarin significantly reduced thrombus weight and length, and apoptosis in HUVECs. Our findings indicated that the combination of EGCG and warfarin protects HUVECs from oxidative stress and prevents apoptosis. However, HIF-1α silencing weakened these effects, which indicated that HIF-1α may participate in DVT. Furthermore, HIF-1α silencing significantly up-regulated cell apoptosis and ROS generation, and enhanced VEGF expression and the activation of the PI3K/AKT and ERK1/2 signaling pathways. In conclusion, our results indicate that EGCG combined with warfarin modifies HIF-1α and VEGF to prevent DVT in rabbits through anti-inflammation via the PI3K/AKT and ERK1/2 signaling pathways.

Epigallocatechin 3‐Gallate Inhibitory Effects on Inflammatory Mediators in BV‐2 Microglial Cells

Chronic neuroinflammation has been associated with many neurodegenerative diseases such as Alzheimer's and Parkinson's Disease. The chronic activation of microglia can cause neuronal damage by releasing proinflammatory cytokines and reactive oxygen mediators. Epigallocatechin‐3‐Gallate (EGCG) is a bioactive polyphenol that exhibits significant anti‐inflammatory and neuroprotective properties in green tea extract. The current study aims to examine how EGCG can prevent chronic inflammation by inhibiting proinflammatory responses and oxidative stress induction. If proven to be the case, EGCG may serve as a nutraceutical for preventing neurodegenerative disorders. In this study, we evaluated the anti‐inflammatory activity of EGCG in lipopolysaccharide (LPS) stimulated BV‐2 microglia cells. BV‐2 microglial cells were first stimulated with 1µg/mL LPS for 1hr; then incubated for 24 hrs using different concentrations of EGCG. Cell viability was assessed using resazurin (Alamar™ Blue) indicator dye. In addition, inflammatory cytokines were profiled using QIAGEN mouse inflammatory cytokines multi‐analyte ELISA array, and nitric oxide (NO) release was evaluated using the Griess Reagent Assay. Our results showed that EGCG caused a concentration‐dependent decrease in cell viability of LPS stimulated BV‐2 microglia cells for EGCG concentrations higher than 150 µM. EGCG also significantly inhibited the release of the proinflammatory cytokine IL‐6 and the inflammatory mediator NO production at 150 µM, and 175µM in LPS stimulated BV‐2 microglial cells. In conclusion, EGCG has decreased IL‐6 and NO production, but further research will be performed to elucidate the anti‐inflammatory and antioxidative attributes of EGCG on inflammation‐induced neurodegenerative diseases.

Promotion Effect of EGCG on the Raised Expression of IL-23 through the Signaling of STAT3-BATF2-c-JUN/ATF2

Tea polyphenol of epigallocatechin-3-gallate (EGCG) has been verified to possess multiple biological activities. Interleukin-23 (IL-23) is a heterodimeric cytokine consisting of two subunits of IL-23p19 and IL-12p40, with the functionality in regulating the production of cytokines under physiological or pathological conditions. By serendipity, the raised expression of IL-23 was observed after treating cells with EGCG, whereas the detailed mechanism remains poorly understood. This study was proposed to investigate the signaling related to EGCG-induced IL-23. The raised expression of IL-23 was confirmed primarily by intraperitoneally injecting with different concentrations of EGCG (0, 20, 50, 80 mg/kg) into BALB/c mice, and the raised expression was confirmed by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Results from enzyme-linked immunosorbent assay (ELISA) revealed the increase of IL-23 in serum from 116.09 to 153.90 pg/mL after treating with EGCG. The same results were also observed in RAW264.7 and peritoneal macrophages after treating with EGCG (0, 1, 5, 10, 25 μM) with the increased tendency of IL-23 in cultural medium (7.98 to 25.38 pg/mL for RAW264.7; 3.64 to 260.93 pg/mL for peritoneal macrophages). After preliminary exploration of the signaling related to the increased IL-23, the classical signaling pathways and key transcription factors, such as nuclear factor kappa-B (NF-κB), mitogen-activated protein kinase (MAPK) signaling pathways, and interferon regulatory factor 5 (IRF5), were demonstrated with no relevant contribution. A further study revealed the involvement of the key transcription factor of BATF2, which could antagonistically modulate the transcription and translation of IL-23. The signaling of STAT3-BATF2-c-JUN/ATF2-IL-23 has been further verified in RAW264.7 macrophages using the STAT3 inhibitor of AG490 and the activator of Colivelin TFA. The results indicated that EGCG inhibits the phosphorylation of STAT3 to facilitate the decreased level of BATF2, which contributed to the increased level of IL-23 by the enhancing heterodimerization of c-JUN and ATF2.

Epigallocatechin-3-Gallate Promotes Osteo-/Odontogenic Differentiation of Stem Cells from the Apical Papilla through Activating the BMP-Smad Signaling Pathway.

Stem cells from apical papilla (SCAPs) are desirable sources of dentin regeneration. Epigallocatechin-3-gallate (EGCG), a natural component of green tea, shows potential in promoting the osteogenic differentiation of bone mesenchymal stem cells. However, whether EGCG regulates the odontogenic differentiation of SCAPs and how this occurs remain unknown. SCAPs from immature human third molars (16–20 years, n = 5) were treated with a medium containing different concentrations of EGCG or bone morphogenic protein 2 (BMP2), with or without LDN193189 (an inhibitor of the canonical BMP pathway). Cell proliferation and migration were analyzed using a CCK-8 assay and wound-healing assay, respectively. Osteo-/odontogenic differentiation was evaluated via alkaline phosphatase staining, alizarin red S staining, and the expression of osteo-/odontogenic markers using qPCR and Western blotting. We found that EGCG (1 or 10 μM) promoted the proliferation of SCAPs, increased alkaline phosphatase activity and mineral deposition, and upregulated the expression of osteo-/odontogenic markers including dentin sialophosphoprotein (Dspp), dentin matrix protein-1 (Dmp-1), bone sialoprotein (Bsp), and Type I collagen (Col1), along with the elevated expression of BMP2 and phosphorylation level of Smad1/5/9 (p < 0.01). EGCG at concentrations below 10 μM had no significant influence on cell migration. Moreover, EGCG-induced osteo-/odontogenic differentiation was significantly attenuated via LDN193189 treatment (p < 0.01). Furthermore, EGCG showed the ability to promote mineralization comparable with that of recombinant BMP2. Our study demonstrated that EGCG promotes the osteo-/odontogenic differentiation of SCAPs through the BMP–Smad signaling pathway.

Epigallocatechin-3-Gallate Promotes the in vitro Maturation and Embryo Development Following IVF of Porcine Oocytes.

PurposeEpigallocatechin-3-gallate (EGCG) is a major ingredient of catechin polyphenols and exerts protective effects because of its strong antioxidant properties. As far as we know, there is still a lack of systematic research on the effects of EGCG on the in vitro maturation (IVM) and in vitro fertilization (IVF) of porcine oocytes. The present study aimed to determine the effects of EGCG on the IVM and IVF of porcine oocytes.MethodsPorcine oocytes were treated with different concentrations of EGCG (5, 10 and 20 µM), and the cumulus cell expansion, oocyte maturation rate, reactive oxygen species (ROS), glutathione (GSH) and malondialdehyde (MDA) levels, total antioxidant capacity were determined. The mRNA expression levels of oxidative stress- and apoptosis-associated genes were determined by quantitative real-time PCR. The cleavage rate and blastocyst rate of oocytes after 10 μM EGCG treatment during IVM and IVF were also evaluated.ResultsEGCG at 5, 10 and 20 μM significantly promoted cumulus cell expansion, and EGCG at 10 μM increased the oocyte maturation rate. EGCG (10 μM) treatment reduced the ROS and MDA levels, while increased the antioxidant capacity and GSH concentrations in the mature oocytes. The qRT-PCR results showed that EGCG treatment up-regulated the mRNA expression of catalase, glutathione peroxidase and superoxide dismutase in the mature oocytes. In addition, EGCG treatment also decreased the mRNA expression levels of Bax and caspase-3 and increased the Bcl-2 mRNA expression level in the mature oocytes. In addition, the cleavage rate and blastocyst rate of oocytes treated with 10 μM EGCG during IVM and IVF were significantly higher than those of the control group.ConclusionOur results suggest that EGCG promotes the in vitro maturation and embryo development following IVF of porcine oocytes. The protective effects of EGCG on the oocytes may be associated with its antioxidant and anti-apoptosis properties.

Effect of epigallocatechin-3-gallate on tumor necrosis factor-alpha production by human gingival fibroblasts stimulated with bacterial lipopolysaccharide: An in vitro study.

Background:Evidence shows that epigallocatechin-3-gallate (EGCG) in green tea has anti-inflammatory effects.Aim:This study assessed the effect of EGCG on the production of tumor necrosis factor-alpha (TNF-α) as an inflammatory cytokine in periodontitis, which produced by human gingival fibroblasts (HGFs) stimulated with lipopolysaccharide (LPS) of Porphyromonas gingivalis.Materials and Methods:In this study, HGFs were cultured and subjected to LPS and EGCG. Cell viability of different concentrations of EGCG (10, 25, 50, 75, and 100 μM) and LPS (1, 10, 20, and 50 μg/mL) was assessed using methyl-thiazole-tetrazolium (MTT) assay. Then, the best concentrations of EGCG and P. gingivalis LPS were used simultaneously and separately to assess the production of TNF-α by HGFs using the enzyme-linked immunosorbent assay (ELISA). Assessments were done at 1, 3, and 5 days. Data were read using the ELISA reader and analyzed by the SPSS through two-way ANOVA.Results:LPS at 1, 10, and 20 and EGCG at 10.25 and 50 μM showed the least cytotoxicity in MTT assay. ELISA showed EGCG alone decreased the production of TNF-α in all days, except 10 μM on day 1. 1, 10, and 20 μg/mL LPS increased the output of TNF-α on days 1 and 3 while reducing it on day 5. The combination of EGCG and LPS showed a decrease of TNF-α in all days except on day 5 that revealed an increase in the production of TNF-α at 25 and 50 μM EGCG.Conclusion:In the combination use of EGCG and LPS, EGCG shows anti-inflammatory effects by decreasing the production of TNF-α by HGFs stimulated with P. gingivalis.

Epigallocatechin-3-gallate (EGCG) addition as an antioxidant in a cryo-diluent media improves microscopic parameters, and fertility potential, and alleviates oxidative stress parameters of buffalo spermatozoa

The disparity between the endogenous antioxidants concentration and free radicals in spermatozoa results in reactive oxygen species (ROS) generation. In this prospect, epigallocatechin-3-gallate (EGCG) preserves vigorous antioxidant features. Current study explored the influence of EGCG in a cryo-diluent media on microscopic parameters, oxidative stress parameters, and fertility potential of buffalo spermatozoa during cryopreservation. Concisely, collected semen from three donor bulls for four times were then evaluated for volume, motility, concentrations and then dilution in a cryo-diluent media with different concentrations of EGCG (EGCG-0 = control; EGCG-50 = 50 μM, EGCG-100 = 100 μM, EGCG-200 = 200 μM, and EGCG-300 = 300 μM) at 37 °C, cooled to 4 °C in 2 h, equilibrated for 4 h at 4 °C, and cryopreserved. At post-thawing, Computer-Assisted Sperm motion Analysis motilities (total and progressive, %) and rapid velocity (%), plasma membrane functionality, supravital plasma membrane integrity, and mitochondrial potential (%) were found higher (P < 0.05) in EGCG-200, and EGCG-300 than control, whereas average-path, straight-line, and curved-linear velocities (μm/sec), and acrosome integrity (%) were recorded higher in EGCG-300 than control. Further, comet length (μm), and tail length (μm), LPO (lipid peroxidation, μM/mL), and apoptosis-like changes (%) in spermatozoa were significantly decreased in EGCG-300 than control. Seminal plasma antioxidant enzymes activities (glutathione peroxidase, U/mL, and superoxide dismutase, U/mL) were increased with EGCG-300 than control. Moreover, EGCG-300 addition in a cryo-diluent media improves the fertility potential (%) of buffalo spermatozoa. In a nutshell, the inclusion of EGCG-300 in a cryo-diluent media enhances post-thaw microscopic parameters, and fertility potential, whereas decreases oxidative stress parameters in buffalo spermatozoa.

Epigallocatechin-3-gallate (EGCG) reduces apoptosis of preantral follicles through the phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) signaling pathway after in vitro culture of sheep ovarian tissue

The aims of this study were to analyze the effects of different concentrations of epigallocatechin-3-gallate (EGCG) on the primordial follicle survival and development after in vitro culture of ovarian tissue, and to verify the possible involvement of the phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) pathway in the EGCG actions in the sheep ovary. Ovarian fragments were fixed for histological analysis (fresh control) or cultured in α-minimum essential medium alone (α-MEM+: control medium) or with different concentrations of EGCG (0.01; 0.1; 1; 10 or 100 μg/mL) for 7 days. Inhibition of PI3K activity was performed in fragments cultured with 1 μg/mL EGCG plus LY294002. Thereafter, immunohistochemistry was performed to evaluate the expression of cleaved caspase-3 and AKT phosphorylation (p-AKT). The results showed that 1 μg/mL EGCG maintained the follicular survival similar (P > 0.05) to that of the fresh control and higher (P < 0.05) than that of the α-MEM+ and other EGCG treatments. No difference (P > 0.05) in the follicular activation was observed. However, both follicle and oocyte diameters increased after in vitro culture with 1 μg/mL EGCG compared to other treatments (P < 0.05), except for 10 μg/mL EGCG (P > 0.05). After PI3K inhibition, there was an increase (P < 0.05) of the follicular apoptosis and a reduction of p-AKT immunolocalization. In conclusion, EGCG at 1 μg/mL reduces apoptosis of preantral follicles through the PI3K/AKT pathway after in vitro culture of sheep ovarian tissue.

(-)-Epigallocatechin-3-gallate induces interferon-λ2 expression to anti-influenza A virus in human bronchial epithelial cells (BEAS-2B) through p38 MAPK signaling pathway.

Background(-)-Epigallocatechin-3-gallate (EGCG), a major component of green tea, has been found to inhibit the influenza virus. However, the mechanism of EGCG anti-influenza virus effect needs to be further explored.MethodsBEAS-2B cells were treated with different concentrations of EGCG or were treated with EGCG for different times. CCK8 assay was used to detect the cell viability, and quantitative real time polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay were employed to measure the interferon (IFN)-λ2 mRNA and protein expression levels. The phospho-p38 mitogen-activated protein kinase (P-p38 MAPK), phospho-extracellular signal-regulated kinase (P-ERK), and phospho-c-Jun N-terminal kinase (P-JNK) expression were tested by western blot. Then, p38 MAPK, ERK, and JNK inhibitor were used to study the effect of p38 MAPK, ERK, and JNK signaling pathways on IFN-λ2 expression. The BEAS-2B cells were treated with EGCG, EGCG and IFN λ2 neutralizing antibody or control antibody for 12 h, and were infected with influenza A virus (IAV) (H1N1) for 1 h. After 12 h, nucleoprotein (NP) mRNA and protein expression levels of H1N1 were assessed by qRT-PCR and western blot.ResultsThe IFN-λ2 mRNA and protein expression levels in BEAS-2B cells were up-regulated after EGCG (treatment in time- and dose-dependent manners the concentration range from 0 to 50 µg/mL had no cytotoxicity). Meanwhile, the P-p38 MAPK, P-ERK, and P-JNK expression levels were up-regulated. IFN-λ2 mRNA and protein expression was inhibited after p38 MAPK inhibitor pre-treatment, but not by ERK and JNK inhibitors. Furthermore, the expression of H1N1 NP gene and protein decreased after EGCG pre-treatment, while IFN-λ2 neutralizing antibody attenuated the effect of EGCG inhibiting the expression of H1N1 NP gene and protein.ConclusionsEGCG inhibited IAV H1N1 by inducing the expression of IFN-λ2 in BEAS-2B cells through the p38 MAPK signaling pathway.

Epigallocatechin gallate inhibits cell growth and regulates miRNA expression in cervical carcinoma cell lines infected with different high-risk human papillomavirus subtypes.

The aim of the present study was to investigate the inhibitory effects of the polyphenol epigallocatechin-3-gallate (EGCG) on the growth of cervical carcinoma cell lines infected with different high-risk human papillomavirus (HPV) subtypes, as well as the associated regulation of microRNA (miR) expression. Cell proliferation was measured using an MTT assay. The effects of 7 different concentrations of EGCG (100, 80, 60, 40, 20, 10 and 0 µg/ml) on HeLa cell proliferation were assessed. HeLa cell growth was significantly inhibited by EGCG in a dose- and time-dependent manner (P<0.05), and the IC50 was 90.74 and 72.74 µg/ml at 24 and 48 h, respectively. The expression of miR-210, miR-29a, miR-203 and miR-125b in HeLa (HPV16/18+), SiHa (HPV16+), CaSki (HPV16+) and C33A (HPV-) cell lines was measured using quantitative polymerase chain reaction analysis. In CA33 cells, miR-203 (all P<0.001) and miR-125b (P<0.01 and <0.0001) were significantly downregulated by EGCG, and miR-210 was significantly upregulated with 40 and 60 µg/ml EGCG (P<0.0001). miR-125b was significantly downregulated (P<0.001 and <0.0001), and miR-210 and miR-29 were significantly upregulated by ≤80 µg/ml EGCG in HeLa cells (all P<0.0001). In CaSki cells, miR-210, miR-29a (all P<0.001) and miR-125b (P<0.01–0.0001) were significantly upregulated by EGCG. In SiHa cells, miR-125b (both P<0.001) and miR-203 (P<0.01 and <0.0001) were significantly upregulated by EGCG. In conclusion, the results of the present study suggest that EGCG suppresses cervical carcinoma cell growth, possibly via regulating the expression of miRs, suggesting their potential as therapeutic targets for the control and prevention of cervical cancer. Additionally, EGCG may be considered a novel anti-cervical cancer drug in the future.

Encapsulation of (─)-epigallocatechin-3-gallate (EGCG) in solid lipid nanoparticles for food applications

A nanoparticlulate delivery system was prepared for developing a food grade carrier for the major bioactive constituent in green tea; (─)-epigallocatechingallate (EGCG), in order to protect it against degradation during storage and digestion under simulated gastrointestinal pH conditions. EGCG-loaded solid lipid nanoparticles (EGCG-SLNs) were produced by the hot homogenization method. The lipid matrix used in the production consisted of pure cocoa butter. A combination of sodium stearoyl-2-lactylate (SSL) and mono- and diglycerides (MDG) was applied as a surfactant blend. The nanoparticles loaded with different concentrations of EGCG had an average particle size in the range of 108–122 nm. A maximal encapsulation efficiency of 68.5% was obtained. The produced food grade SLNs successfully protected the encapsulated EGCG along the storage period as well as under the adverse conditions at neutral pH values. The developed system offers good potential for enriching food products with EGCG.

Epigallocatechin-3-gallate affects the proliferation, apoptosis, migration and invasion of tongue squamous cell carcinoma through the hippo-TAZ signaling pathway

The purpose of the present study was to investigate the mechanism by which epigallocatechin-3-gallate (EGCG) inhibits the biological behaviors of the tongue squamous cell carcinoma (TSCC) through the Hippo-tafazzin (TAZ) signaling pathway and to provide insights into molecular targeted therapy in TSCC. CAL27 and SCC15 cells were treated with different concentrations of EGCG for 24 h. Cell proliferation was determined using Cell-Counting Kit-8 and EdU assays. Cell apoptosis was evaluated by flow cytometry. Cell migration and invasion were measured using scratch and Transwell assays, respectively. Furthermore, protein levels of associated target genes were detected using a western blot assay. It was demonstrated that EGCG affected biological behaviors of CAL27 and SCC15 cells in concentration- and time-dependent manners. In addition, EGCG decreased the protein levels of TAZ, LATS1, MOB1 and JNK. Overexpression of TAZ alleviated the effect of EGCG on CAL27 cells. Furthermore, the combination of EGCG and simvastatin inhibited the proliferation, migration and invasion, and promoted apoptosis significantly compared with single treatment in CAL27 cells. The results of the present study suggested that EGCG affects proliferation, apoptosis, migration and invasion of TSCC through the Hippo-TAZ signaling pathway.

Epigallocatechin‐3‐gallate Suppressed Methylglyoxal‐Induced Apoptosis in SH‐SY5Y Cells

Glycation is an endogenous process that generates a class of compounds known as advanced glycation end products (AGEs). AGEs are found in age‐related intracellular protein deposits of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease. Green tea polyphenols have a role in the reversal of age‐related loss of neuronal plasticity and recovery after neuronal lesions. There are several polyphenolic catechins in green tea, where epigallocatechin‐3‐gallate (EGCG) is both the most abundant and most biologically active form. EGCG could exhibit protective effects against AGEs‐induced injury in neuronal cells through its antioxidative properties, suggesting a beneficial role for this tea catechin against neurodegenerative diseases. In the current study, efforts were made to investigate the effects of EGCG on MG‐induced apoptosis. SH‐SY5Y cell were treated with different concentrations of EGCG followed by treatment with MG. Our data exhibited that both the cleaved PARP and cleaved‐caspase‐3 were downregulated by EGCG, and the MG‐induced reduction of Bcl‐2 expression were reversed by EGCG treatment. Moreover, EGCG could protect cells from death via reverse MG‐induced promoting of ERK/JNK pathway, and associated with reduced p53 accumulation. Our results implicate that EGCG could suppress MG‐induced apoptosis.Support or Funding InformationThis study was supported by NSC 100‐2313‐B‐309‐003 to A‐C ChengThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

The Effect of (−)‐Epigallocatechin‐3‐gallate (EGCG) on Cell Viability and Protein Expression in Human Pancreatic Cancer Cells (Panc‐1) and Rat Osteosarcoma Cell Cultures (UMR 106‐01 BSP)

Traditional treatment of cancer includes surgery, radiation, and chemotherapy. There are many studies regarding how cancer cells respond to various treatments. However, not many studies have been done on how cancer cells respond to natural compounds such as (−)‐Epigallocatechin‐3‐gallate (EGCG), a potent ingredient of green tea. In this study, it was hypothesized that EGCG inhibits cell viability of both human pancreatic (Panc‐1) and rat osteosarcoma, UMR 106‐01 BSP (UMR) cell cultures, inhibits the expression of Panc‐1 CXCR‐4 and VEGF‐A proteins in Panc‐1 cell cultures, and the expression of the oncoproteins MYC and PD‐L1 in UMR cell cultures. Briefly, Panc‐1 and UMR cell cultures were routinely passaged, treated with different concentrations of EGCG (0, 20, 40, 60, 80 and 100 μM) in 6 well plates, then incubated for 48 hours at 37 degrees Celsius under humidification and 5% carbon dioxide infusion. At the end of the experiment, all cells were counted using the LUNA Automated Cell Counter (Logos Biosystems). Upon electrophoresis of experimental samples, Western blotting technique was employed to identify the expression of the Panc‐1 proteins (CXCR‐4, VEGFA) and the UMR oncoproteins (MYC and PD‐L1). F test ANOVA was used to compare variances and all values in the results were expressed as means ±SD. Cell viability results from the Panc‐1 and UMR cell lines showed a decrease at the higher concentrations (80 and 100 μM EGCG). The results of the Western blotting technique showed that the expression of Panc‐1 proteins and UMR oncoproteins were inhibited when treated with EGCG. The cell viability results suggest that EGCG may provide a therapeutic effect on both cancer cells lines at higher concentrations. Results of the Western blotting suggest that EGCG may exert its inhibitory effects on Panc‐1 and UMR cell viability through the regulation of Panc‐1 proteins and UMR oncoproteins.Support or Funding InformationThis work was made possible by support from the Ryckman Endowment Student Research Fund of the Biology Department of La Sierra University.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Effects of epigallocatechin-3-gallate on bovine oocytes matured in vitro.

ObjectiveEpigallocatechin-3-gallate (EGCG) is a major ingredient of catechin polyphenols and is considered one of the most promising bioactive compounds in green tea because of its strong antioxidant properties. However, the protective role of EGCG in bovine oocyte in vitro maturation (IVM) has not been investigated. Therefore, we aimed to study the effects of EGCG on IVM of bovine oocytes.MethodsBovine oocytes were treated with different concentrations of EGCG (0, 25, 50, 100, and 200 μM), and the nuclear and cytoplasmic maturation, cumulus cell expansion, intracellular reactive oxygen species (ROS) levels, total antioxidant capacity, the early apoptosis and the developmental competence of in vitro fertilized embryos were measured. The mRNA abundances of antioxidant genes (nuclear factor erythriod-2 related factor 2 [NRF2], superoxide dismutase 1 [SOD1], catalase [CAT], and glutathione peroxidase 4 [GPX4]) in matured bovine oocytes were also quantified.ResultsNuclear maturation which is characterized by first polar body extrusion, and cytoplasmic maturation characterized by peripheral and cortical distribution of cortical granules and homogeneous mitochondrial distribution were significantly improved in the 50 μM EGCG-treated group compared with the control group. Adding 50 μM EGCG to the maturation medium significantly increased the cumulus cell expansion index and upregulated the mRNA levels of cumulus cell expansion-related genes (hyaluronan synthase 2, tumor necrosis factor alpha induced protein 6, pentraxin 3, and prostaglandin 2). Both the intracellular ROS level and the early apoptotic rate of matured oocytes were significantly decreased in the 50 μM EGCG group, and the total antioxidant ability was markedly enhanced. Additionally, both the cleavage and blastocyst rates were significantly higher in the 50 μM EGCG-treated oocytes after in vitro fertilization than in the control oocytes. The mRNA abundance of NRF2, SOD1, CAT, and GPX4 were significantly increased in the 50 μM EGCG-treated oocytes.ConclusionIn conclusion, 50 μM EGCG can improve the bovine oocyte maturation, and the protective role of EGCG may be correlated with its antioxidative property.

Epigallocatechin-3-gallate

PurposeThe purpose of the study was to explore the possibility of utilization of epigallocatechin-3-gallate (EGCG) as a novel preservative in cheese. EGCG has strong antioxidant and antimicrobial properties and may be commercially exploited as a natural antioxidant and preservative in cheese like products which are highly susceptible to lipid oxidation.Design/methodology/approachThe study was conducted to evaluate the possibility of using EGCG as a natural antioxidant and preservative in cheese. Kalari, a hard and dry cheese, was used as a model and treated with different concentrations of EGCG (0, 0.05 and 0.10 per cent) and aerobically packaged within low-density polyethylene pouches and assessed for oxidative stability and storage quality under refrigerated (4 ± 1°C) conditions.FindingsEGCG showed a significant (p < 0.05) effect on the lipid oxidative stability as the treated products exhibited significantly (p < 0.05) lower thiobarbituric acid reactive substances (milligram malonaldehyde/kg) values. A significant (p < 0.05) effect was also observed on the microbiological characteristics of the products, as treated products showed significantly (p < 0.05) lower values for total plate count (log cfu/g), psychrophilic count (log cfu/g), yeast and mould count (log cfu/g) and free fatty acid (% oleic acid) values. Coliforms (log cfu/g) were not detected throughout the storage period. Significantly (p < 0.05) higher scores were observed for various sensory parameters of the treated products. EGCG successfully improved the oxidative stability and storage quality of Kalari.Originality/valueThe results suggest that EGCG might be useful to the dairy industry as an efficient alternative to synthetic antioxidants and preservatives.

Evaluation of Epigallocatechin-3-gallate Modified Collagen Membrane and Concerns on Schwann Cells.

Collagen is an essential component of the extracellular matrix (ECM) and is a suitable material for nerve repair during tissue remodeling for fracture repair. Epigallocatechin-3-gallate (EGCG), an extract of green tea, shows various biological activities that are beneficial to nerve repair. Here, we developed modified collagen containing different concentrations of EGCG (0.0064%, 0.064%, and 0.64%, resp.) to induce Schwann cell proliferation and differentiation. Cell Counting Kit-8 test, live/dead assay, and SEM showed that collagen cross-linked by EGCG induced Schwann cell proliferation. Real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blotting revealed that EGCG-modified collagen induced Schwann cell differentiation and downregulated reactive oxygen species (ROS) levels by downregulating the MAPK P38 signaling pathway. Our results indicate that collagen cross-linked with an appropriate concentration of EGCG induces the proliferation and differentiation of Schwann cells. The EGCG-modified collagen membrane may be applicable for nerve repair and guided tissue regeneration applications.