Reduction of photodynamic damage of blood vessels in the protected region by(–)-epigallocatechin gallate

Photodynamic therapy(PDT)has been increasingly used in the clinical treatment of neoplastic,inflammatory and infectious skin diseases.However,the generation of reactive oxygen species(ROS)may induce undesired side effects in normal tissue surrounding the treatment lesion,which is a big challenge for the clinical application of PDT.To date,(–)-Epigallocatechin gallate(EGCG)has been widely proposed as an antiangiogenic and antitumor agent for the protection of normal tissue from ROS-mediated oxidative damage.This study evaluates the regulation ability of EGCG for photodynamic damage of blood vessels during hematoporphyrin monomethyl ether(Hemoporfin)-mediated PDT.The quenching rate constants of EGCG for the triplet-state Hemoporfin and photosensitized 1O2 generation are determined to be 6.8×10^(8)M^(−1)S^(−1),respectively.The vasoconstriction of blood vessels in the protected region treated with EGCG hydrogel after PDT is lower than that of the control region treated with pure hydrogel,suggesting an efficiently reduced photodamage of Hemoporfin for blood vessels treated with EGCG.This study indicates that EGCG is an efficient quencher for triplet-state Hemoporfin and 1O2,and EGCG could be potentially used to reduce the undesired photodamage of normal tissue in clinical PDT.

Therapeutic effects of epigallocatechin and epigallocatechin gallate on the allergic reaction ofα_(s1)-casein sensitized mice

To investigate the anti-α_(s1)-casein allergy mechanism of two tea-derived polyphenols,epigallocatechin(EGC)and epigallocatechin gallate(EGCG),BALB/c mice were sensitized and challenged withα_(s1)-casein and nutritional intervention was given by EGC and EGCG during the sensitization provocation phase.The main evaluation indexes used were levels of mast cell proteases,histamine,and specific antibody immunoglobulin E(IgE),as well as cytokine secretion and pathological observation.The results showed that both EGC and EGCG significantly reduced levels of mast cell protease,histamine,specific IgE antibodies,and Th2 cytokines in allergic mice.The histopathology results showed that both EGC and EGCG markedly reduced the degree of lesions in the intestine,thymus,spleen,and lung.The conclusions from this study can provide a theoretical basis for the mechanism by which tea polyphenols regulate food allergens.

Effect of epigallocatechin gallate in green tea on preventing lens opacity and αB-crystallin aggregation in rat model of diabetes

AIM:To evaluate the effect of epigallocatechin gallate(EGCG) in preventing lens opacity and the aggregation of lens αB-crystallin in model rats of diabetes mellitus(DM).METHODS:This experimental study included Wistar rats for DM as in vivo models and divided into 5 groups.The treatment groups were administered EGCG by orally for 20d and were then assessed for their degree of lens opacity with binocular microscope and lens αB-crystallin expression from Western blot analyze.RESULTS:Pearson correlation test and regression analysis on EGCG exposure and final random blood sugar(RBS) obtained a significance level of P<0.05.EGCG exposure can significantly lower RBS with an R~2 of 0.5634(56.34%).The same analysis on EGCG exposure and the degree of lens opacity obtained a significance level of P<0.05 and increased exposure to EGCG can significantly lower the degree of lens opacity with an R~2 of 0.8577(85.77%).Correlation analysis between EGCG and the expression of lens αB-crystallin can be concluded that the higher the EGCG exposure administered,the higher the native lens αB-crystallin expression and the lower the aggregate lens αB-crystallin expression.There was also significant effect in which every 1 mg/kg body weight dose of EGCG can increase the native lens αB-crystallin expression by 0.0063 and decrease the aggregate lens αB-crystallin expression by 0.0076.CONCLUSION:The administration of EGCG at a dose of 300,600,and 1200 mg shows a significant effect on preventing lens opacity and aggregation of αB-crystallin in diabetic rat models and this research could be a biomolecular prevention of cataract.

EGCG对淀粉基乳液凝胶性质的影响

为探究表没食子儿茶素没食子酸酯(epigallocatechin gallate,EGCG)对蜡质玉米淀粉凝胶及其稳定的乳液凝胶性质的影响,该研究在制备淀粉凝胶的过程中添加EGCG,研究EGCG对淀粉凝胶结构性质、流变性质和乳化性质的影响。结果发现,添加EGCG可导致淀粉凝胶网络结构孔径增加、剪切应力和黏弹性增加。通过激光共聚焦荧光显微镜和冷冻电镜观察发现,不同EGCG添加量的淀粉凝胶能够在油滴表面形成不同的包覆结构,其中,EGCG添加量为12%的淀粉凝胶在油滴表面形成了均匀的凝胶网络包覆结构,能够抑制油滴聚结,形成的乳液凝胶在室温下贮藏1 a仍然具有良好的物理稳定性。这表明添加EGCG的淀粉凝胶能够作为乳化剂和凝胶剂用于构建食品级乳液凝胶,对食品和药品工业开发新型淀粉基乳液递送载体具有潜在应用价值。

Epigallocatechin gallate content change of the fresh tea leaf homogenates extracted by different methods in extraction and preservation

The fresh leaves of China green tea, Camellia sinensis, were collected from Fuyang, Zhejiang Province, China, in April. The tea polyphenols was extracted by four different methods （homogenized with distilled water at room temperature, homogenized with 0.3% citric acid （w/v） at room temperature, 5- min boiling and homogenized with distilled water at room temperature, homogenized with 85℃ distilled water）, and after preserving at room temperature, the change of the Epigallocatechin gallate （EGCG） contents of the extracts was investigated. Results indicated that the EGCG content of homogenate extracted with 85℃ distilled water was the highest before the extract was preserved, followed by that of the extract homogenized with 0.3% citric acid at room temperature. During preservation, EGCG content changed obviously. The EGCG contents of homogenates extracted with distilled water at room temperature and 85℃ distilled water declined quickly and separately reduced to 21.52% and 54.6% of their initial contents after preservation for 12 h. The EGCG contents extracted by 0.3% citric acid （w/v） solvent at room temperature and 5- min boiling/homogenized with distilled water at room temperature declined relatively slowly ,and separately reduced to 76.9% and 85.16% of their initial contents after preservation for 12 h. It was also found that the citric acid can prevent the degradation of EGCG and the extract solution color is light green

Structural shift of gut microbiota during chemopreventive effects of epigallocatechin gallate on colorectal carcinogenesis in mice

AIM To investigate the effect of epigallocatechin gallate(EGCG) on structural changes of gut microbiota in colorectal carcinogenesis.METHODS An azoxymethane(AOM)/dextran sodium sulfate(DSS)-induced colitis mouse model was established. Fortytwo female FVB/N mice were randomly divided into the following three groups: group 1(10 mice, negative control) was treated with vehicle, group 2(16 mice, positive control) was treated with AOM plus vehicle, and group 3(16 mice, EG) was treated with AOM plus EGCG. For aberrant crypt foci(ACF) evaluation, the colons were rapidly took out after sacrifice, rinsed with saline, opened longitudinally, laid flat on a polystyrene board, and fixed with 10% buffered formaldehyde solution before being stained with 0.2% methylene blue in saline. For tumor evaluation, the colon was macroscopically inspected and photographed, then the total number of tumors was enumerated and tumor size measured. For histological examination, the fixed tissues were paraffin-embedded and sectioned at 5 mm thickness. Microbial genomic DNA was extracted from fecal and intestinal content samples using a commercial kit. The V4 hypervariable regions of 16 S r RNA were PCR-amplified with the barcoded fusion primers. Using the best hit classification option, the sequences from each sample were aligned to the RDP 16 S r RNA training set to classify the taxonomic abundance in QIIME. Statistical analyses were then performed.RESULTS Treatment of mice with 1% EGCG caused a significant decrease in the mean number of ACF per mouse, when compared with the model mice treated with AOM/DSS(5.38 ± 4.24 vs 13.13 ± 3.02, P < 0.01). Compared with the positive control group, 1% EGCG treatment dependently decreased tumor load per mouse by 85%(33.96 ± 6.10 vs 2.96 ± 2.86, respectively, P < 0.01). All revealed that EGCG could inhibit colon carcinogenesis by decreasing the number of precancerous lesions as well as solid tumors, with reduced tumor load and delayed histological progression of CRC. During the cancerization, the diversity of gut microbiota increased, potential carcinogenic bacteria such as Bacteroides were enriched, and the abundance of butyrate-producing bacteria(Clostridiaceae, Ruminococcus, etc.) decreased continuously. In contrast, the structure of gut microbiota was relatively stable during the intervention of EGCG on colon carcinogenesis. Enrichment of probiotics(Bifidobacterium, Lactobacillu, etc.) might be a potential mechanism for EGCG's effects on tumor suppression. Via bioinformatics analysis, principal coordinate analysis and cluster analysis of the tumor formation process, we found that the diversity of gut microbiota increased in the tumor model group while that in the EGCG interfered group(EG) remained relatively stable.CONCLUSION Gut microbiota imbalance might be a potential mechanism for the prevention of malignant transformation by EGCG, which is significant for diagnosis, treatment, prognosis evaluation, and prevention of colorectal cancer.

WPI和WPI-EGCG构建姜黄素纳米乳液的体外消化差异

选择接枝率为3%和4%的乳清分离蛋白(whey protein isolate,WPI)-表没食子儿茶素没食子酸酯((-)-epigallocatechin-3-gallate,EGCG)接枝物为乳化剂构建姜黄素纳米乳液,探究纳米乳液体外模拟消化过程中游离脂肪酸(free fat acid,FFA)释放和消化特性的差异。结果表明,EGCG的结合可能导致WPI分子结构的展开;与WPI相比,WPI-EGCG稳定乳液的界面膜厚度增加了31.6 nm。WPI-EGCG接枝物稳定的乳液的粒径分散度和平均粒径较小,形成的纳米乳液更加稳定,可以更好地促进脂质消化。4%WPI-EGCG稳定的纳米乳液在肠消化120 min后的最终FFA释放率达到85.13%。并且接枝还提高了系统中封装的姜黄素的生物可及性。

EGCG和EGC对微波加热海鲈鱼鱼片功能特性和蛋白氧化的影响

热加工既可以消除食品中的有害微生物,又可以改变肉类食品风味、质地及营养特性和消化特性,而过度热加工会导致蛋白质氧化,对水产品的感官品质和营养价值产生负面影响。为降低蛋白质在热加工中的氧化水平,常加入天然多酚类物质以改善蛋白质的功能特性或营养价值。研究不同浓度表没食子儿茶素没食子酸酯(EGCG)和表没食子儿茶素(EGC)对微波加热的海鲈鱼鱼片感官特性和蛋白氧化的影响。结果表明:EGCG和EGC使微波加热的海鲈鱼鱼片蒸煮损失率显著下降,相比对照组,蒸煮损失率分别降低了47.98%~62.72%,38.54%~58.63%。硬度和咀嚼度下降,羰基含量也下降;水分含量和巯基含量上升。EGCG和EGC显著改善了海鲈鱼鱼片肌肉纤维组织的断裂和聚集情况;降低了鱼片中的酸味感且没有增加其涩味感;EGCG和EGC减少了氮氧化合物类的挥发性气味,促使含硫化合物类气味的生成。

Does combined therapy of curcumin and epigallocatechin gallate have a synergistic neuroprotective effect against spinal cord injury？

Systematic inflammatory response after spinal cord injury （SCI） is one of the factors leading to lesion development and a profound degree of functional loss. Anti-inflammatory compounds, such as curcumin and epigallocatechin gallate （EGCG） are known for their neuroprotective effects. In this study, we investigated the effect of combined therapy of curcumin and EGCG in a rat model of acute SCI induced by balloon compression. Immediately after SCI, rats received curcumin, EGCG, curcumin ＋ EGCG or saline [daily intraperitoneal doses （curcumin, 6 mg/kg; EGCG 17 mg/kg）] and weekly intramuscular doses （curcumin,60 mg/kg; EGCG 17 mg/kg）] for 28 days. Rats were evaluated using behavioral tests （the Basso, Beattie, and Bresnahan （BBB） open-field locomotor test, flat beam test）. Spinal cord tissue was analyzed using histological methods （Luxol Blue-cresyl violet staining） and mmunohistochemistry （anti-glial fibrillary acidic protein, anti-growth associated protein 43）. Cytokine levels （interleukin-1β, interleukin-4, interleukin-2,interleukin-6, macrophage inflammatory protein 1-alpha, and RANTES） were measured using Luminex assay. Quantitative polymerase chain reaction was performed to determine the relative expression of genes （Sort1, Fgf2, Irf5, Mrc1, Olig2, Casp3, Gap43, Gfap, Vegf, NfκB, Cntf） related to regenerative processes in injured spinal cord. We found that all treatments displayed significant behavioral recovery, with no obvious synergistic effect after combined therapy of curcumin and ECGC. Curcumin and EGCG alone or in combination increased axonal sprouting, decreased glial scar formation, and altered the levels of macrophage inflammatory protein 1-alpha, interleukin-1β, interleukin-4 and interleukin-6 cytokines. These results imply that although the expected synergistic response of this combined therapy was less obvious, aspects of tissue regeneration and immune responses in severe SCI were evident.

EGCG调控PI3K/Akt通路对CIS诱导大鼠急性肾损伤的作用机制研究

[目的]探讨表没食子儿茶素没食子酸酯(epigallocatechin-3-gallate,EGCG)对顺铂(cisplatin,CIS)诱导的大鼠急性肾损伤(acute kidney injury,AKI)的保护效果和作用机制,以期为CIS和EGCG的临床用药提供试验基础。[方法]选取40只雄性Wistar大鼠,随机均分为5组。CON和CIS组大鼠灌胃生理盐水;EGCG、CE和抑制剂组大鼠灌胃40 mg/kg EGCG,连续28 d;在第26天,CIS、CE及抑制剂组大鼠腹腔注射7 mg/kg CIS,CON和EGCG组大鼠注射等量生理盐水,此外抑制剂组大鼠腹腔注射5 mg/kg LY294002,连续3 d。第29天采集所有大鼠的肾脏组织,采用HE染色及透射电镜观察肾脏组织病理和细胞线粒体形态变化;利用Western blotting检测各组大鼠肾脏中自噬蛋白(LC3Ⅱ/Ⅰ、ATG5、ATG12、Beclin-1、p62)、通路蛋白(PI3K、Akt、p-PI3K、p-Akt)的表达情况;通过免疫荧光法检测肾脏中自噬相关蛋白LC3Ⅱ/Ⅰ、p62的表达水平。[结果]HE染色及透射电镜结果显示,预处理EGCG缓解了CIS诱导大鼠肾小管上皮细胞坏死和脱落、炎性细胞浸润及细胞线粒体肿胀变性和线粒体嵴断裂。Western blotting结果显示,与CON组相比,CIS组大鼠肾脏中LC3Ⅱ/Ⅰ、ATG5、ATG12、Beclin-1表达量极显著升高(P<0.01),p62、p-PI3K/PI3K和p-Akt/Akt表达量极显著降低(P<0.01),而预处理EGCG组逆转了这一现象。免疫荧光结果显示,与CE组相比,抑制剂组大鼠病理损伤加重,LC3Ⅱ/Ⅰ荧光信号增强,p62荧光信号减弱。[结论]预处理EGCG可以通过激活PI3K/Akt信号通路抑制细胞自噬,从而缓解CIS诱导的大鼠AKI,而这种保护作用可以被PI3K/Akt信号通路抑制剂LY294002抵消。

Epigallocatechin gallate inhibits HBV DNA synthesis in a viral replication-inducible cell line

AIM:To analyze the antiviral mechanism of Epigallocatechin gallate(EGCG)against hepatitis B virus(HBV) replication.METHODS:In this research,the HBV-replicating cell line HepG2.117 was used to investigate the antiviral mechanism of EGCG.Cytotoxicity of EGCG was analyzed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay.Hepatitis B virus e antigen(HBeAg)and hepatitis B virus surface antigen(HBsAg)in the supernatant were detected by enzyme-linked immunosorbent assay.Precore mRNA and pregenomic RNA(pgRNA) levels were determined by semi-quantitative reverse transcription polymerase chain reaction(PCR)assay.The effect of EGCG on HBV core promoter activity was measured by dual luciferase reporter assay.HBV covalently closed circular DNA and replicative intermediates of DNA were quantified by real-time PCR assay.RESULTS:When HepG2.117 cells were grown in the presence of EGCG,the expression of HBeAg was suppressed,however,the expression of HBsAg was not affected.HBV precore mRNA level was also downregulated by EGCG,while the transcription of precore mRNA was not impaired.The synthesis of both HBV covalently closed circular DNA and replicative intermediates of DNA were reduced by EGCG treatment to a similar extent,however,HBV pgRNA transcripted from chromosome-integrated HBV genome was not affected by EGCG treatment,indicating that EGCG targets only replicative intermediates of DNA synthesis.CONCLUSION:In HepG2.117 cells,EGCG inhibits HBV replication by impairing HBV replicative intermediates of DNA synthesis and such inhibition results in reduced production of HBV covalently closed circular DNA.

High glucose reduces Nrf2-dependent cRAGE release and enhances inflammasome-dependent IL-1βproduction in monocytes:the modulatory effects of EGCG

Soluble receptor for advanced glycation end products(sRAGE)acts as a decoy sequestering of RAGE ligands,thus preventing the activation of the ligand-RAGE axis linking human diseases.However,the molecular mechanisms underlying sRAGE remain unclear.In this study,THP-1 monocytes were cultured in normal glucose(NG,5.5 mmol/L)and high glucose(HG,15 mmol/L)to investigate the effects of diabetesrelevant glucose concentrations on sRAGE and interleukin-1β(IL-1β)secretion.The modulatory effects of epigallocatechin gallate(EGCG)in response to HG challenge were also evaluated.HG enhanced intracellular reactive oxygen species(ROS)generation and RAGE expression.The secretion of sRAGE,including esRAGE and cRAGE,was reduced under HG conditions,together with the downregulation of a disintegrin and metallopeptidase 10(ADAM10)and nuclear factor erythroid 2-related factor 2(Nrf2)nuclear translocation.Mechanistically,the HG effects were counteracted by siRAGE and exacerbated by siNrf2.Chromatin immunoprecipitation results showed that Nrf2 binding to the ADAM10 promoter and HG interfered with this binding.Our data reinforce the notion that RAGE and Nrf2 might be sRAGE-regulating factors.Under HG conditions,the treatment of EGCG reduced ROS generation and RAGE activation.EGCG-stimulated cRAGE release was likely caused by the upregulation of the Nrf2-ADAM10 pathway.EGCG inhibited HG-mediated NLRP3 inflammasome activation at least partly by stimulating sRAGE,thereby reducing IL-1βrelease.

Comparison of the impact of epigallocatechin gallate and ellagic acid in an experimental cataract model induced by sodium selenite

AIM：To compare the potential protective effects of epigallocatechin gallate（EGCG） and ellagic acid（EA） in an experimental cataract model.●METHODS：Twenty-eight Spraque-Dawley rat pups were assigned into four groups.All the rats,except for those in the control group,were injected subcutaneously sodium selenite to induce experimental cataract on the postpartum ninth day,and between 10 th and 14 th days.Rats in the sham,EGCG,and EA groups were intraperitoneally administered 50 mg/（kg·d） saline solution,50 mg/（kg·d） EGCG and 200 mg/（kg·d） EA,respectively.The reduced glutathione（GSH） and malondialdehyde（MDA） levels,total antioxidant status（TAS） and total oxidant status（TOS） in lens supernatants were measured.RESULTS：The mean cataract gradings in EGCG and EA groups were found to be significantly lower than that in sham group（P〈0.001）.The mean GSH levels and TASs in EGCG and EA groups were significantly higher than that in sham group while mean MDA levels and TOSs in EGCG and EA groups were significantly lower than that in the sham group（P〈0.001）.CONCLUSION：EGCG and EA have protective effects on cataract development via the inhibition of oxidative stress.

表没食子儿茶素没食子酸酯(EGCG)对碘酸钠诱导的干性年龄相关性黄斑变性模型大鼠视网膜氧化损伤的抑制作用

目的探讨表没食子儿茶素没食子酸酯(EGCG)对碘酸钠诱导的干性年龄相关性黄斑变性(AMD)模型大鼠视网膜氧化损伤的抑制作用。方法取SPF级雄性SD大鼠36只随机分为空白对照组、碘酸钠组、碘酸钠+EGCG组,每组12只。碘酸钠组和碘酸钠+EGCG组大鼠按体质量尾静脉注射50 mg·kg-1碘酸钠,建立干性AMD模型,空白对照组大鼠给予等量生理盐水。造模后碘酸钠+EGCG组大鼠右眼玻璃体内注射4μL 0.5 g·L^(-1) EGCG,空白对照组、碘酸钠组大鼠右眼给予等量生理盐水,21 d后将大鼠处死、取材并检测指标。HE染色观察各组大鼠视网膜组织病理形态变化,检测大鼠视网膜组织中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)含量,Western blot检测大鼠视网膜Nrf2、NADPH氧化还原酶(NQO1)、Ⅰ型血红素氧化酶(HO-1)蛋白表达水平,实时荧光定量PCR检测大鼠视网膜Nrf2、NQO1、HO-1 mRNA相对表达量。结果HE染色结果显示,与空白对照组相比,碘酸钠组大鼠视网膜全层均损伤,视网膜色素上皮层细胞损伤明显,外核层排列紊乱,呈波浪状改变;与碘酸钠组相比,碘酸钠+EGCG组大鼠视网膜各层损伤得到改善。与空白对照组相比,碘酸钠组大鼠视网膜组织SOD、GSH-Px含量均降低,差异均有统计学意义(均为P<0.01),MDA含量升高,差异有统计学意义(P<0.01);与碘酸钠组相比,碘酸钠+EGCG组大鼠视网膜组织SOD、GSH-Px含量均升高,差异均有统计学意义(均为P<0.01),MDA含量降低,差异有统计学意义(P<0.01)。Western blot检测结果显示,与空白对照组相比,碘酸钠组大鼠视网膜Nrf2、NQO1、HO-1蛋白相对表达量均升高,差异均有统计学意义(均为P<0.01);与碘酸钠组相比,碘酸钠+EGCG组大鼠视网膜Nrf2、NQO1、HO-1蛋白相对表达量均升高,差异均有统计学意义(均为P<0.05)。实时荧光定量PCR检测结果显示,与空白对照组相比,碘酸钠组大鼠视网膜Nrf2、NQO1和HO-1 mRNA相对表达量均升高,差异均有统计学意义(均为P<0.05);与碘酸钠组相比,碘酸钠+EGCG组大鼠视网膜Nrf2、NQO1、HO-1 mRNA相对表达量均升高,差异均有统计学意义(均为P<0.05)。结论EGCG主要是通过上调Nrf2表达,进而激活下游产物NQO1、HO-1表达,提高SOD、GSH-Px含量,降低MDA含量,从而增强机体清除氧自由基的能力,抑制碘酸钠诱导的干性AMD大鼠视网膜氧化损伤。

表没食子儿茶素没食子酸酯-牛骨蛋白(EGCG-BBP)偶联物对乳化肉丸蛋白结构及氧化稳定性的影响

为探究表没食子儿茶素没食子酸酯-牛骨蛋白(Epigallocatechin-3-gallate-Bovine bone protein,EGCG-BBP)对乳化肉制品蛋白结构及贮藏氧化稳定性的影响,本文研究不同EGCG-BBP添加量对生肉糜中肌原纤维蛋白(Myofibrillar protein,MP)的理化性质、结构特性以及对肉丸氧化特性的影响。结果表明:当EGCG-BBP添加量为0.8%时,肉糜中MP的巯基含量最高,达4.06 nmol/mg蛋白,且羰基含量及表面疏水性最低,能够有效提升乳化肉制品的抗氧化能力。由红外光谱分析表明,与未添加EGCG-BBP组相比,添加共价物肉糜中MP的酰胺A带峰值所对应的波数明显增大,说明MP的二级结构会随之发生改变;荧光光谱显示,随贮藏时间延长,对照组中MP的最强荧光波长发生显著红移,但随EGCG-BBP浓度的增加,红移程度显著降低,表明添加EGCGBBP能够改变MP的三级结构。此外,乳化肉丸贮藏过程中的氧化指标分析表明,添加0.8%EGCG-BBP能显著降低肉丸的过氧化值(PV)和硫代巴比妥酸值(TBARs),从而提高其氧化稳定性。综上所述,EGCG-BBP能够显著改变MP的二、三级结构,且具有良好的抗氧化性能,在提升乳化肉制品品质方面具有很大的应用潜力,为肉品抗氧化型乳化剂的应用提供新的选择。

Epigallocatechin gallate inhibits dimethylhydrazine-induced colorectal cancer in rats

BACKGROUND Epigallocatechin gallate(EGCG)is a polyhydroxy phenolic compound extracted from tea and its antitumor effect has received widespread attention.We explored the inhibitory effect of EGCG on dimethylhydrazine(DMH)-induced colorectal cancer(CRC)using a rat model,predicted the interaction between EGCG and CRC target genes using a database,and explained the EGCG associated target pathways and mechanisms in CRC.AIM To understand the inhibitory mechanisms of EGCG on CRC cell proliferation and identify its pharmacological targets by network pharmacology analysis.METHODS DMH(40 mg/kg,s.c.,twice weekly for eight weeks)was used to induce CRC in rats.After model establishment,the rats were administered with EGCG(50,100,or 200 mg/kg,p.o.,once daily for eight weeks)and killed 12 and 20 wk after the start of the experiment.Formation of aberrant crypt foci and tumor was studied by histological analysis.Using network pharmacology analysis,candidate and collective targets of EGCG and CRC were identified,and Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes analyses were used to predict the pathways altered by EGCG.RESULTS At week 12,high-dose EGCG treatment significantly reduced the tumor formation rate,total number of tumors,cancerous and non-cancerous tumors,tumor volume,ascites formation,and aberrant crypt foci count.At week 20,all three doses of EGCG were effective.Seventy-eight collective targets of EGCG and CRC were identified,of which 28 genes were dysregulated in CRC.Kyoto Encyclopedia of Genes and Genomes and GO analyses showed that the dysregulated genes were enriched in hsa05210(CRC),hsa04115(p53 signaling pathway),and hsa04151(PI3K-Akt signaling pathway),GO:0043124(negative regulation of I-kappaB kinase/NF-kappaB signaling pathway),GO:0043409(negative regulation of mitogen-activated protein kinase cascade),and GO:2001244(positive regulation of intrinsic apoptotic signaling pathway)respectively.CONCLUSION EGCG inhibits the formation of DMH-induced CRC by regulating key pathways involved in tumorigenesis.

超声预处理对酪蛋白酸钠-EGCG相互作用及其复合物抗氧化活性的影响

采用荧光分光光度计、紫外分光光度计等研究了超声预处理对酪蛋白酸钠-表没食子儿茶素没食子酸酯((-)-epigallocatechin-3-gallate,EGCG)相互作用及其复合物抗氧化活性的影响,并从蛋白质结构变化上探究了其作用机理。结果表明:EGCG对酪蛋白酸钠内源荧光的淬灭形式为静态淬灭。超声预处理后酪蛋白酸钠同EGCG之间的结合常数由9.51×10^(3) L·(mol/L)^(-1)提高至7.48×10^(4) L·(mol/L)^(-1),结合位点数由0.871提高至1.067,分别提高了7.87倍和22.50%,说明二者结合能力增强。同酪蛋白酸钠相比,超声预处理酪蛋白酸钠无规卷曲含量增加,内源荧光强度和表面疏水性指数增强,结构的改变促进了其同EGCG的结合。酪蛋白酸钠-EGCG复合物的形成可以抑制EGCG的降解、提高其抗氧化稳定性,而超声预处理能增强复合物的这一作用。

EGCG联合力学刺激对成骨细胞增殖与分化的影响

目的探究表没食子儿茶素没食子酸酯(EGCG)联合力学刺激(MS)对成骨细胞增殖分化的诱导作用。方法实验分为对照组、EGCG组、MS组、EGCG+MS组;采用MTT法检测EGCG对成骨细胞的增殖活性;采用茜素红染色法检测EGCG对成骨细胞矿化的影响;采用四点弯曲力学加载装置向细胞施加力学载荷,应用碱性磷酸酶(ALP)测试盒检测ALP活性;采用免疫印迹法检测成骨细胞runt相关转录因子-2(Runx-2)和I型胶原蛋白(Col-I)的表达。结果浓度为30μmol/L的EGCG对成骨细胞的增殖作用最明显,差异有统计学意义(P<0.05);EGCG组的矿化结节数量多于对照组,与单一EGCG组和MS组相比,EGCG+MS组ALP活性明显提高,Runx-2和Col-I蛋白的表达增加(P<0.05)。结论EGCG联合力学刺激增强了成骨细胞ALP活性以及Runx-2和Col-I蛋白的表达,成骨细胞的增殖与分化能力增加。

基于网络药理学探讨EGCG对顺铂所致大鼠急性肾损伤的保护作用

目的基于网络药理学和体内动物模型实验,评价表没食子儿茶素没食子酸酯(epigallocatechin-3-gallate,EGCG)对顺铂(cisplatin,CIS)诱导的大鼠急性肾损伤(acute kidney injury,AKI)的保护效果。方法通过TCMSP、Gene Cards、OMIM网站收集EGCG、AKI作用靶点,取交集后构建蛋白质-蛋白质互作网络(PPI),Cytoscape 3.9.1对交集靶点进行可视化分析,筛选出关键靶点,通过DAVID数据库进行KEGG和GO富集分析。32只雄性Wistar大鼠,随机分为对照组(CON组)、EGCG组、顺铂组(CIS组)和CIS+EGCG组。CON组和CIS组灌胃生理盐水,EGCG组和CIS+EGCG组灌胃EGCG(40 mg/kg),连续28 d,第26天CIS组、CIS+EGCG组腹腔注射CIS(7 mg/kg),第29天收集血液和组织。检测血清尿素氮(BUN)、肌酐(SCr)水平;苏木素-伊红(HE)染色观察肾病理变化;TUNEL检测肾组织细胞凋亡情况;Western Blot、qRT-PCR及免疫组化验证分析结果。结果网络药理学筛选出87个EGCG与AKI交集基因,25个核心靶点,通过PI3K/AKT等信号通路和多种生物过程影响AKI的发展;EGCG预处理明显降低AKI大鼠血清中BUN、SCr水平,改善AKI大鼠肾病变,缓解AKI大鼠肾组织凋亡;Western Blot、qRT-PCR及免疫组化结果表明预处理EGCG可激活PI3K/AKT通路。结论基于以上研究结果,推测出EGCG可能通过激活PI3K/AKT信号通路进而缓解CIS诱导的大鼠AKI。

Neuroprotective effect of epigallocatechin-3-gallate on hemisection-induced spinal cord injury in rats

Epigallocatechin-3-gallate （EGCG）, a naturally occurring compound in green tea, has been widely used as an antioxidant agent. In the present study, model rats with acute spinal cord injury were intraperitoneally injected with 25, 50, and 100 mg/kg EGCG, and spinal cord ultrastructure, oxidative stress reaction, inflammatory factors, and apoptosis-associated gene expression were observed. Results showed that EGCG attenuated neuronal and axonal injury 24 hours post injury. It also decreased serum intedeukin-113, tumor necrosis factor-a, and intercellular adhesion molecule-1 release, and decreased apoptosis-associated gene expression. Furthermore, it increased the level of the superoxide anion （O2-）, superoxide dismutase, and B-cell lymphoma/leukemia-2, and reduced malondialdehyde levels. Furthermore, it reduced the expression of the pro-apoptotic protein Bax. Noticeably, EGCG at the 100 mg/kg dosage exhibited similar effects as methylprednisolone sodium succinate, which has been frequently used for clinical acute spinal cord injury. The results demonstrated that EGCG can significantly inhibit inflammation, suppress oxidation, and reduce apoptosis in acute spinal cord injury.