Enhancement of Annexin V in response to combination of epigallocatechin gallate and quercetin as a potent arrest the cell cycle of colorectal cancer / Aumento da anexina V em resposta à combinação de galato de epigalocatequina e quercetina como uma potente parada do ciclo celular do câncer colorretal

Abstract Colorectal cancer (CRC) is one of the most common cancers leading to comorbidities and mortalities globally. The rational of current study was to evaluate the combined epigallocatechin gallate and quercetin as a potent antitumor agent as commentary agent for therapeutic protocol. The present study investigated the effect of epigallocatechin Gallate (EGCG) (150mg) and quercetin (200mg) at different proportions on proliferation and induction of apoptosis in human colon cancer cells (HCT-116). Cell growth, colonogenic, Annexin V in addition cell cycle were detected in response to phytomolecules. Data obtained showed that, the colony formation was inhibited significantly in CRC starting from the lowest concentration tested of 10 µg/mL resulting in no colonies as visualized by a phase-contrast microscope. Data showed a significant elevation in the annexin V at 100 µg/mL EGCG(25.85%) and 150 µg/mL quercetin (48.35%). Moreover, cell cycle analysis showed that this combination caused cell cycle arrest at the G1 phase at concentration of 100 µg/mL (72.7%) and 150 µg/mL (75.25%). The combined effect of epigallocatechin Gallate and quercetin exert antiproliferative activity against CRC, it is promising in alternative conventional chemotherapeutic agent.

Resumo O câncer colorretal (CCR) é um dos cânceres mais comuns, levando a comorbidades e mortalidade em todo o mundo. O racional do presente estudo foi avaliar a combinação de galato de epigalocatequina e quercetina como um agente antitumoral potente como agente de comentário para protocolo terapêutico. O presente estudo investigou o efeito de galato de epigalocatequina (EGCG) (150 mg) e quercetina (200 mg) em diferentes proporções na proliferação e indução de apoptose em células de câncer de cólon humano (HCT-116). O crescimento celular, colonogênico, anexina V, além do ciclo celular foram detectados em resposta a fitomoléculas. Os dados obtidos mostraram que a formação de colônias foi inibida significativamente no CRC a partir da concentração mais baixa testada de 10 µg/mL, resultando em nenhuma colônia conforme visualizado por um microscópio de contraste de fase. Os dados mostraram uma elevação significativa na anexina V a 100 µg/mL de EGCG (25,85%) e 150 µg/mL de quercetina (48,35%). Além disso, a análise do ciclo celular mostrou que essa combinação causou parada do ciclo celular na fase G1 na concentração de 100 µg/mL (72,7%) e 150 µg/mL (75,25%). O efeito combinado da epigalocatequina galato e quercetina exerce atividade antiproliferativa contra o CCR, é promissor como agente quimioterápico alternativo convencional.

Synergistic antimicrobial potential of EGCG and fosfomycin against biofilms associated with endodontic infections

Abstract Objective This study aimed to evaluate the cytotoxicity and synergistic effect of epigallocatechin gallate (EGCG) and fosfomycin (FOSFO) on biofilms of oral bacteria associated with endodontic infections. Methodology This study determined minimum inhibitory and bactericidal concentration (MIC/MBC) and fractionated inhibitory concentration (FIC) of EGCG and FOSFO against Enterococcus faecalis, Actinomyces israelii, Streptococcus mutans, and Fusobacterium nucleatum. Monospecies and multispecies biofilms with those bacteria formed in polystyrene microplates and in radicular dentin blocks of bovine teeth were treated with the compounds and control chlorhexidine (CHX) and evaluated by bacterial counts and microscopy analysis. Toxicity effect of the compounds was determined on fibroblasts culture by methyl tetrazolium assays. Results The combination of EGCG + FOSFO demonstrated synergism against all bacterial species, with an FIC index ranging from 0.35 to 0.5. At the MIC/FIC concentrations, EGCG, FOSFO, and EGCG+FOSFO were not toxic to fibroblasts. EGCG+FOSFO significantly reduced monospecies biofilms of E. faecalis and A. israelli, whereas S. mutans and F. nucleatum biofilms were eliminated by all compounds. Scanning electron microscopy of multispecies biofilms treated with EGCG, EGCG+FOSFO, and CHX at 100x MIC showed evident biofilm disorganization and substantial reduction of extracellular matrix. Confocal microscopy observed a significant reduction of multispecies biofilms formed in dentin tubules with 84.85%, 78.49%, and 50.6% of dead cells for EGCG+FOSFO, EGCG, and CHX at 100x MIC, respectively. Conclusion EGCG and fosfomycin showed a synergistic effect against biofilms of oral pathogens related to root canal infections without causing cytotoxicity.

Enhancement of Annexin V in response to combination of epigallocatechin gallate and quercetin as a potent arrest the cell cycle of colorectal cancer / Aumento da anexina V em resposta à combinação de galato de epigalocatequina e quercetina como uma potente parada do ciclo celular do câncer colorretal

Colorectal cancer (CRC) is one of the most common cancers leading to comorbidities and mortalities globally. The rational of current study was to evaluate the combined epigallocatechin gallate and quercetin as a potent antitumor agent as commentary agent for therapeutic protocol. The present study investigated the effect of epigallocatechin Gallate (EGCG) (150mg) and quercetin (200mg) at different proportions on proliferation and induction of apoptosis in human colon cancer cells (HCT-116). Cell growth, colonogenic, Annexin V in addition cell cycle were detected in response to phytomolecules. Data obtained showed that, the colony formation was inhibited significantly in CRC starting from the lowest concentration tested of 10 µg/mL resulting in no colonies as visualized by a phase-contrast microscope. Data showed a significant elevation in the annexin V at 100 µg/mL EGCG(25.85%) and 150 µg/mL quercetin (48.35%). Moreover, cell cycle analysis showed that this combination caused cell cycle arrest at the G1 phase at concentration of 100 µg/mL (72.7%) and 150 µg/mL (75.25%). The combined effect of epigallocatechin Gallate and quercetin exert antiproliferative activity against CRC, it is promising in alternative conventional chemotherapeutic agent.

O câncer colorretal (CCR) é um dos cânceres mais comuns, levando a comorbidades e mortalidade em todo o mundo. O racional do presente estudo foi avaliar a combinação de galato de epigalocatequina e quercetina como um agente antitumoral potente como agente de comentário para protocolo terapêutico. O presente estudo investigou o efeito de galato de epigalocatequina (EGCG) (150 mg) e quercetina (200 mg) em diferentes proporções na proliferação e indução de apoptose em células de câncer de cólon humano (HCT-116). O crescimento celular, colonogênico, anexina V, além do ciclo celular foram detectados em resposta a fitomoléculas. Os dados obtidos mostraram que a formação de colônias foi inibida significativamente no CRC a partir da concentração mais baixa testada de 10 µg/mL, resultando em nenhuma colônia conforme visualizado por um microscópio de contraste de fase. Os dados mostraram uma elevação significativa na anexina V a 100 µg/mL de EGCG (25,85%) e 150 µg/mL de quercetina (48,35%). Além disso, a análise do ciclo celular mostrou que essa combinação causou parada do ciclo celular na fase G1 na concentração de 100 µg/mL (72,7%) e 150 µg/mL (75,25%). O efeito combinado da epigalocatequina galato e quercetina exerce atividade antiproliferativa contra o CCR, é promissor como agente quimioterápico alternativo convencional.

Enhancement of Annexin V in response to combination of epigallocatechin gallate and quercetin as a potent arrest the cell cycle of colorectal cancer

Abstract Colorectal cancer (CRC) is one of the most common cancers leading to comorbidities and mortalities globally. The rational of current study was to evaluate the combined epigallocatechin gallate and quercetin as a potent antitumor agent as commentary agent for therapeutic protocol. The present study investigated the effect of epigallocatechin Gallate (EGCG) (150mg) and quercetin (200mg) at different proportions on proliferation and induction of apoptosis in human colon cancer cells (HCT-116). Cell growth, colonogenic, Annexin V in addition cell cycle were detected in response to phytomolecules. Data obtained showed that, the colony formation was inhibited significantly in CRC starting from the lowest concentration tested of 10 µg/mL resulting in no colonies as visualized by a phase-contrast microscope. Data showed a significant elevation in the annexin V at 100 µg/mL EGCG(25.85%) and 150 µg/mL quercetin (48.35%). Moreover, cell cycle analysis showed that this combination caused cell cycle arrest at the G1 phase at concentration of 100 µg/mL (72.7%) and 150 µg/mL (75.25%). The combined effect of epigallocatechin Gallate and quercetin exert antiproliferative activity against CRC, it is promising in alternative conventional chemotherapeutic agent.

Resumo O câncer colorretal (CCR) é um dos cânceres mais comuns, levando a comorbidades e mortalidade em todo o mundo. O racional do presente estudo foi avaliar a combinação de galato de epigalocatequina e quercetina como um agente antitumoral potente como agente de comentário para protocolo terapêutico. O presente estudo investigou o efeito de galato de epigalocatequina (EGCG) (150 mg) e quercetina (200 mg) em diferentes proporções na proliferação e indução de apoptose em células de câncer de cólon humano (HCT-116). O crescimento celular, colonogênico, anexina V, além do ciclo celular foram detectados em resposta a fitomoléculas. Os dados obtidos mostraram que a formação de colônias foi inibida significativamente no CRC a partir da concentração mais baixa testada de 10 µg/mL, resultando em nenhuma colônia conforme visualizado por um microscópio de contraste de fase. Os dados mostraram uma elevação significativa na anexina V a 100 µg/mL de EGCG (25,85%) e 150 µg/mL de quercetina (48,35%). Além disso, a análise do ciclo celular mostrou que essa combinação causou parada do ciclo celular na fase G1 na concentração de 100 µg/mL (72,7%) e 150 µg/mL (75,25%). O efeito combinado da epigalocatequina galato e quercetina exerce atividade antiproliferativa contra o CCR, é promissor como agente quimioterápico alternativo convencional.

Inhibitory effect of epigallocatechin gallate on human Tenon fibroblasts and its mechanism / 中华实验眼科杂志

Objective:To investigate the effect of epigallocatechin gallate (EGCG) on the activation of human Tenon fibroblasts (HTFs) and its mechanism.Methods:Tenon capsule tissues from nine eyes of nine advanced primary open angle glaucoma patients during trabeculectomy were obtained for primary cell culture.HTFs harvested were identified by immunofluorescence staining for vimentin and keratin.Cells at passage 4-6 were used for experiment.Viability of HTFs treated with EGCG at 0, 10, 20, 30, 40, 50, 60, 70 and 80 μmol/L was detected by cell counting kit-8 (CCK-8) assay.The cells were divided into blank control group, transforming growth factor (TGF)-β1-induced group, and EGCG-treated group, which were cultured in normal medium, medium containing 10 ng/ml TGF-β, medium containing 10 ng/ml TGF-β+ 50 μmol/L EGCG, respectively.The proliferation rate of HTFs was detected by BrdU labeling assay.Cell migration was observed by scratch wound healing assay.The expression of α-smooth muscle actin (α-SMA) was measured by immunofluorescence staining.The protein relative expression levels of Smad2/3, phosphoinositide-3-kinase (PI3K), protein kinase B (Akt) as well as the phosphorylated Smad2/3 (p-Smad2/3) and phosphorylated Akt (p-Akt) were measured by western blot.This study was approved by the Ethics Committee of Guangdong Provincial People's Hospital (NO.GDREC2019331H[R1]).Results:The HTFs harvested had spindle shape, grew regularly and were vimentin-positive.CCK-8 assay showed that there was no significant difference in the variability of HTFs treated with EGCG at 10, 20, 30, 40 and 50 μmol/L in comparison with 0 μmol/L EGCG treatment (all at P<0.05). BrdU labeling assay showed that cell proliferation in the TGF-β1-induced group was (66.37±12.65)%, which was significantly higher than (14.75±12.33)% in EGCG-treated group ( P<0.05). Three days after scratch, the relative scratch area in the TGF-β1-induced group was (47.33±12.22)%, which was significantly lower than (92.67±4.04)% in the EGCG-treated group ( P<0.05). Immunofluorescence assay showed that α-SMA fluorescence was significantly enhanced in the TGF-β1-induced group in comparison with the blank control group, which was reduced to blank control group level in EGCG-treated group.Western blot analysis showed that there were significant differences in the relative expression levels of p-Smad2/3, PI3K and p-Akt protein among the various groups ( F=58.820, 121.153, 69.289; all at P<0.001). The relative expressions of p-Smad2/3, PI3K and p-Akt in the TGF-β1-induced group were significantly higher than those in the blank control group, 10 μmol/L and 50 μmol/L EGCG-treated groups (all at P<0.05). Conclusions:EGCG can suppress TGF-β1-induced HTFs activation through Smad and PI3K/Akt signaling pathways.

Post Oral Administration of Epigallocatechin Gallate from Camelia sinensis Extract Enhances Vascular Endothelial Growth Factor and Fibroblast Growth Factor Expression during Orthodontic Tooth Movement in Wistar Rats.

Background:East Java green tea leaf (Camelia sinensis)possesed active compound such as EpigallocatechinGallate (EGCG) is well known for enhancing the boneremodelling through enhancement of VascularEndothelial Growth Factor (VEGF) and FibroblastGrowth Factors (FGF-2). Remodelling of alveolar boneis very important to obtain optimal Orthodontic ToothMovement (OTM) to align the tooth. Aim: Toinvestigate the expression of VEGF and FGF-2expression during OTM in Wistar rat afteradministration of EGCG from C. sinensis Extract(EGCG-CSE) Wistar rats. Material and Methods: Thisstudy was true experimental study with post-test onlycontrol group design. Twenty eight Wistar rats wererandomly selected and divided into four groupsaccordingly; K- group which did not get both EGCGCSE administration and OTM; K+ group with OTM for14 days, but no EGCG-CSE administration; 1 (T1) with4 days of OTM and 7 days of EGCG-CSEadministration; treatment group 2 (T2) with both 14days OTM and EGCG-CSE administration. Ten g2 force/mm of NiTi close coil spring was installedbetween the upper left molars and cental insicive tomove the molar mesially that induce OTM. All OTMthanimal model were terminated in the 14 days.Maxillary was isolated for immunohistochemistryinvestigation. Tukey Honest Significant Difference(HSD) was done after Analysis of Variance (ANOVA)test to investigate the significant difference betweengroups (p<0.05). Results: The highest positive VEGFexpression was found in the T2 in both area.Meanwhile, the highest positive FGF-2 expression wasfound in the K-group in both area. There weresignificant different of VEGF and FGF-2 expression inboth area between groups except T1 and T2.Conclusion: Post administration of EGCG-CSE canstimulate the VEGF and FGF-2 expression during OTMin Wistar rats.

The use of epigallocatechin gallate prevents white spot lesions during orthodontic treatment with fixed appliances / 中国组织工程研究

BACKGROUND: Plenty of studies have already proved the effective usage of epigallocatechin gallate (EGCG) in clinical treatment. However, no current research has focused on the application of EGCG in preventing white spot lesions (WSLs) during orthodontics treatment with fixed appliances. OBJECTIVE: To study the value of EGCG in the prevention of WSLs during orthodontic treatment with fixed appliances. METHODS: In total 50 patients undergoing orthodontic treatment with fixed appliances were carefully screened and enrolled. Split-mouth design was adopted: the right side of teeth received experimental adhesive (1 g/L EGCG + Adper™ Single Bond 2); the left side of teeth acted as control. All the other clinical procedures and materials used were same. The enamel demineralization index (EDI) and the WSLs prevalence of targeted teeth (16, 11, 46, 26, 31, and 36) were detected at 3, 6, and 12 months during the treatment, and the percentage of bracket bonding failure was calculated for each group. The study protocol was implemented in line with the relevant ethical requirements of Liuzhou People’s Hospital. Patients and their guardians were fully informed of the whole trial procedures. RESULTS AND CONCLUSION: In this trial, the percentage of bracket bonding failure was significantly different between the EGCG group and control group (P > 0.05). After 3 months of treatment, the values of WSLs and EDI had no significant difference between the EGCG group and control group (P > 0.05). However, after 6 months and 12 months treatment, the EGCG group manifested significantly lower WSL and EDI values than the control group (P < 0.05). Therefore, addition of the adhesive containing 1 g/L EGCG has a considerable effect in preventing enamel demineralization and the occurrence of WSLs without influencing the enamel bonding strength, and it has a long-time effect which deserves the clinical expansion.

Epigallocatechin gallate induces gene demethylation to promote acute myeloid leukemia cell apoptosis by regulating p19-p53-p21 signaling pathway / 南方医科大学学报

OBJECTIVE@#To investigate the mechanism by which epigallocatechin gallate (EGCG) induces gene demethylation and promotes the apoptosis of acute myeloid leukemia KG-1 and THP-1 cell lines.@*METHODS@#KG-1 and THP-1 cells treated with 25, 50, 75, 100 or 150 μg/mL EGCG for 48 h were examined for gene methylation using MSP and for cell proliferation using MTT assay. The changes in cell cycle and apoptosis of the two cell lines after treatment with EGCG for 48 h were detected using flow cytometry. The mRNA and protein expressions of DNMT1, CHD5, p19, p53 and p21 in the cells were detected using RT-quantitative PCR and Western blot.@*RESULTS@#EGCG dose-dependently reversed hypermethylation of gene and reduced the cell viability in both KG-1 and THP-1 cells ( < 0.05). EGCG treatment caused obvious cell cycle arrest in G1 phase, significantly increased cell apoptosis, downregulated the expression of DNMT1 and upregulated the expressions of CHD5, p19, p53 and p21 in KG-1 and THP-1 cells ( < 0.05).@*CONCLUSIONS@#EGCG reduces hypermethylation of gene in KG-1 and THP-1 cells by downregulating DNMT1 to restore its expression, which results in upregulated expressions of p19, p53 and p21 and induces cell apoptosis.

Research progress on epigallocatechin-3-gallate in the prevention and treatment of oral cancer / 口腔疾病防治

@#Epigallocatechin-3-gallate (EGCG) has antibacterial, anti-inflammatory, antitumor, and other functions. EGCG and its anticancer mechanism are hot research topics in the prevention and treatment of oral cancer. In this paper, the prevention and treatment effects of EGCG on oral cancer and its anticancer mechanism are reviewed. The results show that EGCG can regulate multiple cell metabolic signaling pathways, such as the G protein coupled receptor signaling pathway, mitogen-activated protein kinase (MAPK), and the Wnt signaling pathway, and it can regulate DNA methylation and act on RNA of oral cancer cells directly or indirectly through the oral cancer cell signal transduction network to inhibit the development of oral precancerous lesions and oral cancer. EGCG combined with 5-fluorouracil can enhance the curative effect and reduce adverse effects and is expected to be a new drug for the prevention and treatment of oral cancer.

Influence of Media Supplementation with Alpha Tocopherol and/or Epigallocatechin Gallate on in vitro Maturation and Subsequent Fertilization of Bovine Oocytes

The present study was planned to determine the effect of Epigallocatechin Gallate (EGCG), Alpha tocopherol and their combination as an antioxidant in TCM-199 media for in vitro maturation (IVM) and in vitro fertilization (IVF) in bovine oocytes. Cumulus-oocyte complexes (COCs) were aspirated from the ovaries derived from slaughter house and in vitro cultured in three groups using TCM-199 supplemented with EGCG @10 μM, Alpha tocopherol @100 μM, and Combination (EGCG @10 μM plus Alpha tocopherol @100 μM). Oocytes of a control group were matured in TCM-199 medium without any treatment. After IVM, cumulus-free oocytes were co-incubated with frozen-thawed spermatozoa for 15–18 h. Compared to no addition, the presence of EGCG @10 μM in medium during IVM significantly (p<0.05) increased the proportion of maturation and fertilization rate. Combination produced significantly higher percentage of in vitro matured bovine oocytes compared to the alpha tocopherol @100 μM alone. The results suggest that EGCG @10 μM in IVM medium had better effect than Alpha tocopherol alone and Combination on in vitro maturation and subsequent fertilization of bovine oocytes.

Screening of anti-oxidant components of Gongcheng Tea by HPLC-UV-FLD and micro-injection systems / 中草药

Objective: A high performance liquid chromatography-ultraviolet-fluorescence (HPLC-UV-FLD) derivative system was established and the anti-oxidant activity of the extract of Gongcheng Tea was preliminarily evaluated by using a micro-injector imaging system combined with a fluorescent probe. Methods: Hydrogen peroxide assay and hydroxyl radical scavenging ability experiments were carried out on the anti-oxidant activity of Gongcheng Tea. The anti-oxidant components of Gongcheng Tea were screened by HPLC-UV-FLD post column derivation system and analyzed by an ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF MS). At the same time, the hydrogen peroxide scavenging activity of tea and active components were tested by a micro-injector imaging system combined with a hydrogen peroxide fluorescent probe (NBCD) in living Drosophila melanogaster. Results: The method was simple and rapid, and three main anti-oxidant compounds, epigallocatechin, epigallocatechin gallate, and epicatechin gallate were found in Gongcheng Tea. And it was found that they could significantly reduce the level of hydrogen peroxide in D. melanogaster. Conclusion: It laid a material basis for the further study of the anti-oxidant activity of Gongcheng Tea, and provided a reference for the screening of anti-oxidants in natural products.

Effect of epigallocatechin gallate on Th1/Th2 immune balance in patients with plaque-type psoriasis / 中华皮肤科杂志

Objective@#To evaluate the effect of epigallocatechin gallate (EGCG) on T helper cell 1 (Th1) and Th2 in psoriasis patients.@*Methods@#A total of 33 patients with plaque-type psoriasis vulgaris were enrolled, and peripheral blood mononuclear cells (PBMC) were isolated and cultured. The appropriate concentration of EGCG was determined by methyl thiazol tetrazolium (MTT) assay. PBMC at exponential growth phase were divided into 2 groups to be treated with EGCG (EGCG group) or not (control group) for 24 hours. Flow cytometry was performed to determine proportions of Th1 and Th2 cells, enzyme-linked immunosorbent assay (ELISA) to detect levels of Th1 (interleukin[IL]-2, interferon[IFN]-γ) and Th2 cytokines (IL-4, IL-10) in the cell culture supernatant, and real-time quantitative RCR (qRT-PCR) to determine the mRNA expression of T-bet (a Th1 transcription factor) and GATA3 (a Th2 transcription factor) . Statistical analysis was carried out by using t test.@*Results@#According to the MTT assay results, EGCG at a non-toxic concentration of 60 μmol/L was chosen for subsequent experiments. Compared with the control group, the EGCG group showed significantly decreased number of Th1 cells (t = 3.43, P = 0.026) , increased number of Th2 cells (t = 6.68, P = 0.026) , and decreased Th1/Th2 ratio (P < 0.05) . The levels of IL-2 and IFN-γ in the culture supernatant of PBMC were both significantly lower in the EGCG group (824.45 ± 101.21 ng/L, 1 623.62 ± 185.56 ng/L respectively) than in the control group (1 568.32 ± 196.45 ng/L, 3 287.63 ± 235.54 ng/L respectively) , while the levels of IL-4 and IL-10 were significantly higher in the EGCG group (389.48 ± 46.63 ng/L, 285.95 ± 53.28 ng/L respectively) than in the control group (225.38 ± 26.92 ng/L, 165.46 ± 32.25 ng/L respectively) . Compared with the control group, the EGCG group showed significantly decreased T-bet mRNA expression (t = 11.99, P < 0.001) , but increased GATA3 mRNA expression (t = 18.62, P < 0.001) .@*Conclusion@#EGCG can reduce the number of Th1 cells, inhibit the production of Th1 cytokines and transcription factors, and increase the number of Th2 cells and the production of Th2 cytokines and transcription factors, followed by the modulation of Th1/Th2 immune imbalance.

Study on Mechanism of Epigallocatechin Gallate Alleviating Myocardial Ischemia-reperfusion Injury by Inhibiting Cardiomyo- cyte Apoptosis / 中国药房

OBJECTIVE： To investigate the protective effect and potential mechanism of epigallocatechin gallate （EGCG） on myocardial ischemia-reperfusion injury. METHODS： H9C2 cardiomyocytes were treated with tert-butyl hydroperoxide （TBHP） to establish ischemia-reperfusion cell model. The cell viability was measured by MTS after pretreated with different doses of EGCG （3.125， 6.25， 12.5， 25， 50， 100， 200 μmol/L）， and the survival rate was calculated. The expression of apoptotic proteins （Bcl-2， Bax） in cardiomyocytes pretreated with different doses of EGCG （100， 200 μmol/L） were detected by Western blotting. Male C57BL/6 mice were randomly divided into sham operation group， model group and EGCG group （5 mg/g）， with 15 mice in each group. Sham operation group and model group were given constant volume of normal saline intragastrically， while EGCG group was given relevant medicine intragastrically， once a day， for consecutive 7 d. Twelve hours after last medication， myocardial ischemia-reperfusion injury model was established by anterior descending coronary artery ligation. The area of myocardial infarction was observed by double staining of Evan’s blue and TTC； the percentage of infarction area to cross-sectional area was calculated；SOD activity and MDA content in serum were determined by WST-1 assay； the expression of apoptotic proteins （Bcl-2， Bax） in myocardial tissue were detected by Western blotting， while the phosphorylation levels of signaling pathway related proteins （PI3K， p-PI3K， Akt， p-Akt） were also detected. RESULTS： Cell test results showed that， compared with control group， survival rate and relative expression of Bcl-2 were decreased significantly in model group， while relative expression of Bax was increased significantly （P＜0.05）. Compared with model group， survival rate of cardiomyocyte in 25， 50， 100， 200 μmol/L EGCG groups as well as relative expression of Bcl-2 in 100， 200  μmol/L EGCG groups were increased significantly， while relative expression of Bax in 100， 200 μmol/L EGCG groups were decreased significantly （P＜0.05）. Animal experiments showed that no ischemia of myocardial tissue and enlargement of cardiac cavity were observed in sham operation group. Myocardial infarction was observed in model group. Compared with sham operation group， percentage of infarction area to cross-sectional area， the serum content of MDA， the relative expression of Bax in myocardial tissue and p-PI3K/PI3K， p-Akt/Akt were increased significantly in model group， while SOD activity and relative expression of Bcl-2 were decreased significantly （P＜0.05）. Compared with model group， myocardial infarction area of mice in EGCG group was reduced， the percentage of infarction area to cross-sectional area， the serum content of MDA， the relative expression of Bax in myocardial tissue and p-PI3K/PI3K， p-Akt/Akt were significantly decreased， the activity of SOD activity and the relative expression of Bcl-2 were increased significantly （P＜0.05）. CONCLUSIONS： EGCG can protect against myocardial ischemia-reperfusion injury， the mechanism of which may be associated with inhibiting the apoptosis of myocardial cells， improving oxidation stress， regulating the expression of apoptotic protein， reducing the phosphorylation level of PI3K/Akt signaling pathway-related proteins.

Protective Effect of Epigallocatechin Gallate on Diabetic Rat Hearts via TGF-β1/Smad3 Signaling Pathway / 中国医科大学学报

Objective To investigate the effects of Epigallocatechin gallate (EGCG) on cardiac protection in diabetic rats and the expression of TGF-1/Smad3 signaling pathway. Methods The influence of clean level 40 SD rats, weight 200-220 g, divided into four random groups:control (Sham) group, diabetic cardiomyopathy model (DC) group, EGCG group, and metformin positive control group (Met).Post 8 weeks of high-fat-diet administration, the rats were injected intraperitoneally with STZ to establish the diabetes cardiomyopathy model. Upon successful model establishment, the EGCG group was intraperitoneally injected with EGCG and the cardiac function of the rats was measured after 28 days of drug administration. Then, the pathological results of the myocardial tissue were analyzed. Triglyceride (TG), total cholesterol (TC), glycosylated hemoglobin (HbA1 c), and blood glucose (FBG) concentrations were also measured. Further, the concentrations of superoxide dismutases (SOD), malondialdehyde (MDA), catalase (CAT), and glutathione peroxidase (GPX) in serum were measured by ELISA. The expression of TGF-β1 and Smad3 in kidney tissues of the rats was measured by Western blotting analysis. Results EGCG could reduce the glucose, lipid, and MDA levels in the blood of the diabetic rats, enhance cardiac systolic and diastolic functions, inhibit TGF-β1 and Smad3 protein expression, enhance the activity of SOD, CAT and GPX, and reduce myocardial tissue fibrosis. Conclusion EGCG can protect diabetic rat hearts by improving metabolic disorder, and its mechanism may be related to the oxidative-stress mediated by the TGF-β1/Smad3 signaling pathway.

The epigallocatechin gallate derivative Y reverses drug resistance mediated by the ABCB1 transporter both and

Previously, we reported that Y, a new epigallocatechin gallate derivative, is efficacious in reversing doxorubicin (DOX)--mediated resistance in hepatocellular carcinoma BEL-7404/DOX cells. In this study, we evaluated the efficacy of Y in reversing drug resistance both and by determining its effect on the adenosine triphosphate-binding cassette protein B1 transporter (ABCB1 or P-glycoprotein, P-gp). Our results showed that Y significantly sensitized cells overexpressing the ABCB1 transporter to anticancer drugs that are ABCB1 substrates. Y significantly stimulated the adenosine triphosphatase activity of ABCB1. Furthermore, Y exhibited a higher docking score as compared with epigallocatechin gallate inside the transmembrane domain of ABCB1. In addition, in the nude mouse tumor xenograft model, Y (110 mg/kg, intragastric administration), in combination with doxorubicin (2 mg/kg, intraperitoneal injection), significantly inhibited the growth of BEL-7404/DOX cell xenograft tumors, compared to equivalent epigallocatechin gallate. In conclusion, Y significantly reversed ABCB1-mediated multidrug resistance and its mechanisms of action may result from its competitive inhibition of the ABCB1 drug efflux function.

Effect of epigallocatechin gallate on Th1/rh2 immune balance in patients with plaque-type psoriasis / 中华皮肤科杂志

Objective To evaluate the effect of epigallocatechin gallate (EGCG) on T helper cell 1 (Th1) and Th2 in psoriasis patients.Methods A total of 33 patients with plaque-type psoriasis vulgaris were enrolled,and peripheral blood mononuclear cells (PBMC) were isolated and cultured.The appropriate concentration of EGCG was determined by methyl thiazol tetrazolium (MTT) assay.PBMC at exponential growth phase were divided into 2 groups to be treated with EGCG (EGCG group) or not (control group) for 24 hours.Flow cytometry was performed to determine proportions of Th 1 and Th2 cells,enzyme-linked immunosorbent assay (ELISA) to detect levels of Th1 (interleukin [IL]-2,interferon [IFN]-γ) and Th2 cytokines (IL-4,IL-10) in the cell culture supernatant,and real-time quantitative RCR (qRT-PCR) to determine the mRNA expression of T-bet (a Th1 transcription factor) and GATA3 (a Th2 transcription factor).Statistical analysis was carried out by using t test.Results According to the MTT assay results,EGCG at a non-toxic concentration of 60 μmol/L was chosen for subsequent experiments.Compared with the control group,the EGCG group showed significantly decreased number of Th1 cells (t =3.43,P =0.026),increased number of Th2 cells (t =6.68,P =0.026),and decreased Th1/Th2 ratio (P ＜ 0.05).The levels of IL-2 and IFN-γin the culture supernatant of PBMC were both significantly lower in the EGCG group (824.45 ± 101.21 ng/L,1 623.62 ± 185.56 ng/L respectively) than in the control group (1 568.32 ±196.45 ng/L,3 287.63 ± 235.54 ng/L respectively),while the levels of IL-4 and IL-10 were significantly higher in the EGCG group (389.48 ± 46.63 ng/L,285.95 ± 53.28 ng/L respectively) than in the control group (225.38 ± 26.92 ng/L,165.46 ± 32.25 ng/L respectively).Compared with the control group,the EGCG group showed significantly decreased T-bet mRNA expression (t =11.99,P ＜ 0.001),but increased GATA3 mRNA expression (t =18.62,P ＜ 0.001).Conclusion EGCG can reduce the number of Th1 cells,inhibit the production of Th 1 cytokines and transcription factors,and increase the number of Th2 cells and the production of Th2 cytokines and transcription factors,followed by the modulation of Th 1/Th2 immune imbalance.

Protective effects of epigallocatechin gallate on depressive mice induced by chronic unpredictable mild stress / 中草药

Objective: To observe the protective effects of epigallocatechin gallate (EGCG) on the depression. Methods: The depressive mice model induced by chronic unpredictable mild stress (CUMS) was established. The behavioristics of the mice exposed to various stress were tested form the open-field, body weight, sucrose preference, forced swimming and tail suspension test for investigating the antidepressant effect of EGCG. Meanwhile, serum CORT, ACTH levels, and the hippocampus MDA, SOD and GSH-Px were estimated. The mRNA expression of indoleamine 2,3-dioxygenase (IDO), IL-1β, and IL-6 in the hippocampus tissue were detected by RT-PCR. Results: EGCG improved the behavioristics of depressive mice, however, it ameliorated the serum CORT and ACTH levels. Moreover, EGCG increased the activity of SOD and GSH-Px, and reduced MDA, IDO, IL-6, and IL-1β expression on the mRNA levels in the hippocampus tissue. Conclusion: EGCG could attenuate depressive status of mice, and the underlying mechanism may related to the reduction of serum CORT and ACTH, down-regulation of MDA, IL-1β, IL-6, and IDO, and up-regulation of SOD and GSH-Px in hippocampus.

Protective effects of Epigallocatechin gallate on liver ischemia reperfusion injury in rats / 中华肝胆外科杂志

Objective To investigate the protective roles of Epigallocatechin gallate (EGCG) on liver ischemia reperfusion injury (IRI) in rats.Methods 30 healthy male SD rats were selected and equally and randomly divided into 3 groups.Sham group,IRI group and IRI-EGCG group were established to construct 70％ liver IRI rat model.Drinking water with 0.4 mg/ml EGCG was administered for 2 weeks before the experiment in IRI-EGCG group.HE staining was performed to evaluate the injury.Transaminases in serum were investigated to assess liver injury.p-p85 and p-AKT was detected by Western-blot assay.qPCR was carried out to study the mRNA expression of TNF-α,IL-6 and IL-1β in liver tissue.The secretion of TNF-α,IL-6 and IL-1 β in serum was examined with ELISA assay.Results EGCG pretreatment reduced ASTand ALT in serum [AST:(550.0 ±66.5) IU/L vs.(220.0 ±63.5) IU/L;ALT:(376.0 ± 25.7) IU/L vs.(158.0 ± 33.1) IU/L,all P ＜ 0.05] and mitigated liver tissue damage.p-p85 and p-AKT increased due to liver IRI,and IRI-EGCG group showed higher expression of p85 and AKT.The proinflammatory cytokines of TNF-α,IL-6 and IL-1 β exhibited a relatively lower mRNA expression in IRI-EGCG group comparing with IRI group.IRI-EGCG group also revealed a decreased secretion of TNF-α,IL-6 and IL-1β in serum [TNF-α:(398.0±33.4) ng/Lvs.(211.0±23.6) ng/L;IL-6:(341.0±27.3) ng/L vs.(187.0±19.6) ng/L;IL-1β:(486.0±43.7) ng/L vs.(352.0±31.5) ng/L;allP＜0.05].Conclusion EGCG pretreatment can enhance IRI-induced activation of PI3K/AKT signaling and reduce the release of proinflammatory cytokines to exert liver protective effects.

Preventive effect of epigallocatechin gallate on hyperoxia-induced bronchopulmonary dysplasia in neonatal g / 国际儿科学杂志

Objective To study the preventive effect of epigallocatechin gallate(EGCG)on hyperoxiainduced bronchopulmonary dysplasia in mice.Methods Eighty-eight Kunming mice born 36-40 hours were randomly divided into four groups:air + saline group (n =22),air + EGCG group (n =22),hyperoxia + saline group(n =22) and hyperoxia + EGCG group(n =22).Drug was administrated once a day by air-driven atomization,till 28 days.Furthermore,the hyperoxia groups were transferred into the room air for recovery after raised in (70 ± 3) ％ oxygen for 28 days.Lung pathological morphology and homogenization ELISA were performed on the 21th,28th and 49th day.The pathological changes of lung tissue radical alveolar counts(RAC) were observed and the levels of interleukin-2 (IL-2),tumor necrosis factor(TNF-α) and myeloperoxidase(MPO) were measured.Results The levels of IL-2 and TNF-α in the air groups were significantly lower than those in hyperoxia group(IL-2:air(1 760.83 ±303.38)pg/g vs hyperoxia(4 251.00 ±644.07) pg/g;TNF-a:air(4 308.83 ±1 114.91) pg/g vs hyperoxia(8301.83 ± 802.26) pg/g) (P ＜ 0.01),and MPO level decreased in hyperoxia +EG-CG group(7 472.83 ± 1 922)U/g,hyperoxia +NS group(4 767.68 ± 1 110.72)U/g,air + EGCG group(3712.68 ± 734.40)U/g,air + NS group(2 711.68 ± 763.39)U/g(P ＜0.05).In the hyperoxia groups,though IL-2 on 21th day and TNF-αt on 49th day were no statistic difference in hyperoxia group and air group,the inflammatory factors in EGCG group were lower than in hyperoxia + NS group.Pathology showed that the RAC count of the air groups increased gradually,while the hyperoxic groups were gradually decreased,however,the EGCG group tend to recover after 21 days recovery in room air.Conclusion Inhalation of EGCG can reduce the level of inflammatory response in lung tissue of hyperoxia exposed mice.EGCG attenuated lung injury in hyperoxia-exposed mice,and accelerated lung recovery after hyperoxia exposure in experimental animals.EGCG has potential effect to prevent hyperoxia induced bronchopulmonary dysplasia.

Effect of epigallocatechin gallate on ultraviolet B-induced transfer and degradation of melanosomes in mice / 中华皮肤科杂志

Objective To evaluate the effect of tea polyphenol epigallocatechin gallate (EGCG)on ultraviolet B (UVB)-induced skin pigmentation,transfer and degradation of melanosomes in mice,and to explore the role of autophagy in the mechanism of melanosome degradation.Methods A total of 32 ears from 16 female C57/BL6 mice were randomly and equally divided into 4 groups:acetone control group topically treated with acetone solution daily,EGCG group topically treated with 10 g/L EGCG acetone solution daily,UVB irradiation group irradiated with 500 mJ/cm2 UVB once a day and 2 hours later topically treated with acetone solution,UVB + EGCG group irradiated with 500 mJ/cm2 UVB once a day and 2 hours later topically treated with EGCG acetone solution.Ten days later,all the mice were sacrificed,and skin tissue samples were collected from the ears.Transmission electron microscopy was performed to observe ultrastructural changes of melanosomes and autophagosomes,immunohistochemical study to measure expression of protease-activated receptor 2 (PAR2) and microtubule-associated protein light chain 3 (LC3) in the epidermis,and Western blot analysis to determine the protein expression of PAR2,Rasrelated protein Rab27a and LC3 in the epidermis.Results There was a significant difference in the number of melanosomes and autophagosomes among the acetone control group,EGCG group,UVB irradiation group and UVB + EGCG group (H =12.249,13.888,respectively,both P ＜ 0.05).Compared with the acetone control group,the UVB irradiation group showed significantly increased number of melanosomes (1.85 ± 0.32 vs.1.00 ± 0.41,P ＜ 0.05)and autophagosomes (1.94 ± 0.64 vs.1.00 ± 0.46,P ＜ 0.05) in epidermal keratinocytes in mouse skin.Compared with the UVB irradiation group,the UVB + EGCG group showed significantly decreased number of melanosomes (1.30 ± 0.44,P ＜ 0.05),but significantly increased number of autophagosomes (3.03 ± 0.75,P ＜ 0.05).Immunohistochemical study showed a significant difference in the level of PAR2 in the epidermis among the 4 groups (H =18.700,P ＜ 0.05),and the expression of PAR2 was significantly lower in the UVB + EGCG group than in the UVB irradiation group (7.94 ± 4.57 vs.12.54 ± 3.07,Z =2.143,P ＜ 0.05).However,the 4 groups all showed a low level of LC3,and there was no significant difference among the 4 groups (H =5.051,P ＞ 0.05).Western blot analysis revealed significant differences in the protein expression of PAR2 and Rab27a,as well as in the LC3-Ⅱ/LC3-Ⅰ ratio,among the 4 groups (F =18.739,25.967,24.022,respectively,all P ＜ 0.05).Compared with the UVB irradiation group,the UVB + EGCG group showed significantly decreased expression of PAR2 (0.91 ± 0.54 vs.3.12 ± 0.61,P ＜ 0.05) and Rab27a (0.99 ± 0.16 vs.1.42 ± 0.07,P ＜ 0.05),but significantly increased LC3-Ⅱ/LC3-Ⅰ ratio (1.67 ± 0.08 vs.1.24 ± 0.07,P ＜ 0.05).Conclusion Topical EGCG treatment can effectively suppress UVB-induced skin pigmentation,which may be related to the inhibition of melanosome transfer and promotion of melanosome autophagy.