Caffeic acid 3,4-dihydroxyphenethyl ester prevents colorectal cancer through inhibition of multiple cancer-promoting signal pathways in 1,2-Dimethylhydrazine/dextran sodium sulphate mouse model.

OBJECTIVE: To elucidate the potential feature and mechanism of the caffeic acid 3,4-dihydroxyphenethyl ester (CADPE) molecule, which can prevent colorectal cancer (CRC) in the 1,2-Dimethylhydrazine (DMH)/dextran sodium sulphate (DSS)-induced mouse model. METHODS: Institute of cancer research (ICR) male mice were injected with 20 mg/kg DMH for a week. After that, 2% DSS was administered in the drinking water for another 7 d. The CADPE treatment was given to the DMH/DSS induced male mice at three different periods until their sacrifice. Histopathological examination was used for observing the CRC development at colonic mucosa. Immunohistochemistry (IHC), blood cells smearing and crypt damage scoring methods were used for investigating the anti-inflammation feature of CADPE related to CRC. The reversing targets searching method was applied with artificial intelligence (AI), computer-aided drug designing (CADD) and Ingenuity Pathway Analysis (IPA) techniques for predicting the potential targets and mechanism of CADPE highly related to CRC. RESULTS: The data indicated that CADPE inhibited CRC tumor development in the colitis-associated DMH/DSS induced mouse model after giving the early treatment. CADPE also impeded the acute inflammation by decreasing the infiltration of neutrophils significantly during the initial stage of CRC development. Finally, our data showed that CADPE prevented CRC by blocking active sites of three pivotal protein targets including epidermal growth factor receptor (EGFR), extracellular signal-regulated kinase (ERK) and mammalian target of rapamycin (mTOR) in two major cancer development pathways. CONCLUSIONS: CADPE effectively prevented CRC at early stage of tumor germination in the DMH/DSS mouse model highly likely due to its anti-acute inflammation characteristic and the ability of blocking EGFR, ERK and mTOR activities in two highly related CRC developing pathways.

Caffeic Acid Alleviates Memory and Hippocampal Neurogenesis Deficits in Aging Rats Induced by D-Galactose.

Hippocampal neurogenesis occurs throughout life, but it declines with age. D-galactose (D-gal) enhances cellular senescence through oxidative stress leading to neurodegeneration and memory impairment. Caffeic acid (CA) acts as an antioxidant via decreasing brain oxidative stress. This study aims to investigate the advantages of CA in alleviating the loss of memory and neurogenesis production in the hippocampus in aged rats activated by D-gal. Fifty-four male Sprague-Dawley rats were unpredictably arranged into six groups. In the D-gal group, rats were administered D-gal (50 mg/kg) by intraperitoneal (i.p.) injection. For the CA groups, rats received 20 or 40 mg/kg CA by oral gavage. In the co-treated groups, rats received D-gal (50 mg/kg) and CA (20 or 40 mg/kg) for eight weeks. The results of novel object location (NOL) and novel object recognition (NOR) tests showed memory deficits. Moreover, a decline of neurogenesis in the hippocampus was detected in rats that received D-gal by detecting rat endothelial cell antigen-1 (RECA-1)/Ki-67, 5-bromo-2'-deoxyuridine (BrdU)/neuronal nuclear protein (NeuN), doublecortin (DCX) by means of staining to evaluate blood vessel associated proliferating cells, neuronal cell survival and premature neurons, respectively. By contrast, CA attenuated these effects. Our results postulate that CA attenuated the impairment of memory in D-gal-stimulated aging by up-regulating levels of hippocampal neurogenesis.

Pineapple Leaf Phenols Attenuate DSS-Induced Colitis in Mice and Inhibit Inflammatory Damage by Targeting the NF-κB Pathway.

Colitis is not fully curable, although currently, some treatment options are being adopted. In this study, we investigated the effects of pineapple leaf phenols (PLPs), natural phenol products from pineapple leaves, on DSS-induced colitis in mice. The results showed that PLPs dramatically decreased the inflammatory response by inhibiting NF-κB activation and the secretion of pro-inflammatory factors. Moreover, PLPs provided protection against DSS-induced acute colitis by maintaining epithelial integrity. Caffeic and P-coumaric acids had similar effects and could be the active components responsible for PLPs' effect on colitis. These results indicate that the oral administration of PLPs might be considered as a therapeutic strategy in the treatment of patients with colitis. However, further research on clinical applications and the exact effect of PLPs on colitis is required.

Effect of chicoric acid on oxidative stress during myocardial injury in sepsis rats and relationship with Nrf2 signaling pathway / 中华麻醉学杂志

Objective:To evaluate the effect of chicoric acid on oxidative stress during myocardial injury in sepsis rats and the relationship with nuclear factor E2-related factor 2 (Nrf2) signaling pathway.Methods:Forty healthy male Sprague-Dawley rats, aged 8-12 weeks, weighing 220-250 g, were divided into 5 groups ( n=8 each) using a random number table method: control group (group C), lipopolysaccharide (LPS) group (group LPS), LPS+ chicoric acid group (group LPS+ CA), LPS+ Nrf2 inhibitor ML385 group (group LPS+ ML) and LPS+ chicoric acid+ ML385 group (group LPS+ CA+ ML). LPS 15 mg/kg was intraperitoneally injected to induce sepsis.Immediately after intraperitoneal injection of LPS, chicoric acid 10 mg/kg or ML385 15 mg/kg (in dimethyl sulfoxide) was intraperitoneally injected in group LPS+ CA and group LPS+ ML, respectively, and ML385 15 mg/kg and chicoric acid 10 mg/kg were intraperitoneally injected in LPS+ CA+ ML group.The equal volume of dimethyl sulfoxide was given instead in group C. At 48 h after establishment of the model, blood samples were collected from the aorta for measurement of concentration of serum interleukin-6 (IL-6) and the activities of lactate dehydrogenase (LDH) and creatine kinase isoenzyme (CK-MB) (by enzyme-linked immunosorbent assay). The animals were then sacrificed, and myocardial tissues were obtained for microscopic examination of pathological changes (by HE staining), for determination of activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) and contents of reactive oxygen species(ROS) and iron (by colorimetry), for calculation of the ratio of oxidized nicotinamide adenine 2 nucleotides to reduced nicotinamide adenine 2 nucleotides (NAD + /NADH), and for detection of the expression of Nrf2, NADPH quinone oxidoreductase 1 (NQO1), glutathione peroxidase 4 (GPX4) and nicotinamide adenine dinucleotide phosphate oxidase 1 (NOX1) (by Western blot). Results:Compared with C group, the activities of serum LDH and CK-MB and concentration of IL-6 were significantly increased, the contents of ROS and iron and the ratio of NAD + /NADH were increased, activities of GSH-Px and SOD were decreased, expression of Nrf2, NQO1 and GPX4 was down-regulated, and NOX1 expression was up-regulated in the other four groups ( P<0.05). Compared with group LPS, the activities of serum LDH and CK-MB and concentration of IL-6 were significantly decreased, the contents of ROS and iron and the ratio of NAD + /NADH were decreased, activities of GSH-Px and SOD were increased, expression of Nrf2, NQO1 and GPX4 was up-regulated, NOX1 expression was down-regulated ( P<0.05), and the pathological changes of cardiomyocytes were significantly reduced in group LPS+ CA, and the activities of serum LDH and CK-MB and concentration of IL-6 were significantly increased, the ratio of NAD + /NADH were increased, activities of GSH-Px and SOD were decreased, expression of Nrf2, NQO1 and GPX4 was down-regulated, NOX1 expression was up-regulated ( P<0.05), and the pathological changes of cardiomyocytes were accentuated in group LPS+ ML.Compared with group LPS+ CA, the activities of serum LDH and CK-MB and concentration of IL-6 were significantly increased, the contents of ROS and iron and the ratio of NAD + /NADH were increased, activities of GSH-Px and SOD were decreased, expression of Nrf2, NQO1 and GPX4 was down-regulated, NOX1 expression was up-regulated ( P<0.05), and the pathological changes of cardiomyocytes were accentuated in group LPS+ CA+ ML. Conclusion:The mechanism by which chicoric acid reduces myocardial injury in sepsis rats may be related to activating Nrf2 signaling pathway and inhibiting oxidative stress.

Caffeic acid phenethyl ester induces apoptosis in colorectal cancer cells via inhibition of survivin.

Colorectal cancer is one of the most common types of cancer. Drug resistance and drug-induced damage of healthy tissues are major obstacles in cancer treatment. Therefore, to develop efficient anticancer therapy, it is necessary to find compounds that affect tumor cells, but do not exhibit toxicity to healthy cells. Caffeic acid phenethyl ester (CAPE) has been demonstrated to have anticancer properties in many types of cancer. In this study, the cytotoxic and apoptotic effects of CAPE on the RKO colorectal cancer cell line and CCD 841-CoN normal colorectal cell line was investigated. In addition, changes in the survivin expression were determined. According to the results, CAPE decreased cell viability in the RKO cell line in a dose-dependent manner. Likewise, CAPE induced apoptotic cell death in approximately 40% of the RKO cells. Furthermore, CAPE treatment increased the Serine 15 (Ser15) and Serine 46 (Ser46) phosphorylation of p53, while decreased the survivin expression. The results suggested that CAPE induced apoptosis by regulating p53 phosphorylation, leading to inhibition of the survivin expression. In accordance with the results, it is suggested that CAPE might be evaluated as an alternative drug in cancer therapy and further investigation is needed within this scope.

Synthesis of Cinnamyl and Caffeoyl Derivatives of Cucurbitacin-Eglycoside Isolated from Citrullus colocynthis Fruits and their Structures Antioxidant and Anti-inflammatory Activities Relationship.

BACKGROUND: Citrullus colocynthis (L.) Schrad is an important medicinal plant belonging to the family Cucurbitaceae. Cucurbitacin E glucoside (1) was isolated from Citrullus colocynthis fruits. A novel mono-ester of cucurbitacin-E and cinnamyl and caffeoyl-ß-D-glucoside (2 and 3) was synthesized by reaction of cucurbitacin E glucoside with cinnamic and/or caffeic acid in the presence of CHCl2 and K2CO3 with constant stirring with an ice-cooling state for 24h. Mass analyses of the isolated and purified compounds were determined. METHODS: The elemental analysis (C, H, N) suggesting the molecular formulae of the compounds (1-3) to be C38H54O13, C47H60O14, and C47H60O16; respectively. I.R., 1H-NMR, and 13C-NMR analyses were recorded. The median lethal doses (LD50s) of compounds (1-3) in rats were 1262.5, 2500 and 2350 mg/kg b.w., respectively. The anti-inflammatory, total antioxidant, reducing power, anti-reactive oxygen species (ROS) and anti-reactive nitrogen species (RNS) were more pronounced in compound 3 compared to compounds (1-2). This study provides the scientific basis for the anti-inflammatory effects of the isolated cucurbitacin E glucoside (1) and its derivatives (2 and 3) in a t-BHP (tert-butyl hydrogen peroxide)-induced liver damage model. RESULTS: Injection of rats with t-BHP (1.8 mmol/kg) showed a significant increase in plasma alanine transaminases (ALT), aspartate transaminases (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and malondialdehyde (MDA) as well as hepatic tumor Necrosis Factor-α (TNF-α), interleukin- 6 (IL-6) and interleukin-23 (IL-23) when compared with control group. Also, injection of rats with t-BHP showed a significant increase in a liver level of reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST) as compared with control group. Oral administration of cucurbitacin E glucoside (1) and its derivatives (2 and 3) at a concentration of 25 and 50 mg/kg b.wt daily for 5 days showed a significant protection against-induced alteration in liver GSH, SOD, CAT and GST as well as plasma ALT, AST, ALP, LDH and MDA levels. Furthermore, Cucurbitacin E glucoside (1) and its derivatives (2 and 3) inhibited the elevation of proinflammatory cytokines (TNF-α, IL-6, and IL-23) in the livers of t-BHP-treated rat models. CONCLUSION: These results suggested that mechanistic-based evidence substantiating the traditional claims of cucurbitacin E glucoside (1) and its derivatives (2 and 3) to be applied for the treatment of inflammation-related disorders, such as oxidative liver damage and inflammation diseases.

Salvianolic acid A protects rats against cerebral ischemic injury by regulation Wnt/glycogen synthase-kinase-3β/β-catenin signaling pathw ay / 国际脑血管病杂志

Objective To investigate the protective effect of salvianolic acid A (SAA) on permanent focal cerebral ischemia in rats and its possible mechanisms. Methods Fifty-four adult male Sprague-Daw ley rats w ere randomly divided into a sham operation group, a cerebral ischemia group, and a SAA group ( n =18 in each group). A model of permanent middle cerebral artery occlusion w as induced by the intraluminal suture method.At 0 h and 6 h after modeling, the rats of the SAA groups w ere intraperitonealy injected SAA (3 mg/kg). The other groups w ere injected equal volume of saline. At 24 h after modeling, the neurological deficit scores w ere performed. 2,3,5-Triphenyl tetrazolium chloride (TTC) staining w as used to detect cerebral infarction volume. TUNEL staining w as used to detect cel apoptosis. Both immunohistochemical staining and Western blotting w ere used to detect the expressions of Wnt3a, β-catenin, and phosphor-glycogen synthase-kinase-3β(p-GSK-3β) in the ischemic cortex. Results The neurological deficit scores show ed that no neurological deficits w ere observed in the sham operation group (score 0). The neurological deficit score in the SAA group (median and interquartile range) w as significantly low er than that in the cerebral ischemia group (3 [2-3] vs.4 [3-5]; Z = -2.679, P =0.007). No infarcts w ere observed in the sham operation group. The infarct volume in the SAA group w as reduced significantly compared w ith the cerebral ischemia group (79.038 ±10.665 mm 3 vs.212.702 ±8.029 mm 3; t = 24.525, P < 0.001). Very few positive cels w ere observed in the sham operation group. The numbers of TUNEL -positive cels in the SAA group and the cerebral ischemia group w ere 29.667 ±1.366/HP and 63.333 ±0.894/HP, respectively. The former w as significantly less than the latter ( t = 14.115, P < 0.001). Immunohistochemical staining show ed that the number of Wnt3a positive cels in the sham operation group, the cerebral ischemia group, and the SAA group w ere 35.500 ±2.572/HP, 18.056 ±3.765/HP, and 29.000 ±2.376/HP, respectively. There w ere significant differences among the 3 groups ( F = 115.972, P < 0.001), and those in the SAA group w ere significantly more than the cerebral ischemia group ( P < 0.01). The numbers of p-GSK-3βpositive cels in the sham operation group, the model group, and the SAA group w ere 7.944 ±2.127/HP, 37.444 ±3.434/HP, and 11.222 ±1.734/HP, respectively. There w ere significant differences among the three groups (F =730.580, P < 0.001), and those in the SAA group w ere significantly less than the cerebral ischemia group ( P < 0.01). The numbers of β-catenin positive cels in the sham operation group, the cerebral ischemia group, and the SAA group w ere 26.722 ±26.722/HP, 16.556 ±1.854/HP, and 21.333 ± 1.940/HP, respectively. There w ere also significant differences among the 3 groups ( F < 33.385, P <0.01), and those in the SAA group w ere significantly more than the cerebral ischemia group ( P < 0.01). Western blot analysis show ed that Wnt3a expression levels in the sham operated group, the cerebral ischemia group, and the SAA group w ere 1.000 ±0.190, 0.800 ±0.185, and 1.198 ±0.262, respectively. There w ere significant differences among 3 groups ( F = 9.621, P < 0.001), and those in the SAA group w ere significantly higher than the cerebral ischemia group ( P < 0.01). The p-GSK-3βexpression levels in the sham operation group, the cerebral ischemia group, and the SAA group w ere 0.650 ±0.150, 1.290 ± 0.250, and 1.190 ±0.250, respectively. There w ere also significant differences among the 3 groups ( F =19.668, P < 0.001), and those in the SAA group w ere significantly higher than the cerebral ischemia group (P <0.01). The β-catenin expression levels in the sham operation group, the cerebral ischemia group, and the SAA group w ere 1.200 ±0.210, 0.500 ±0.120, and 1.100 ±0.220, respectively. There w ere significant differences among the 3 groups ( F = 33.385, P < 0.001), and those in the SAA group w ere significantly higher than the cerebral ischemia group ( P < 0.01). Conclusions SAA has certain protective effect on permanent cerebral ischemia injury in rats. Its mechanism may be associated w ith the up -regulation of the expression of Wnt3a and β-catenin and the dow n-regulation of the expression of p-GSK-3β.

Simultaneous Determination of Four Active Components in Tobacco Wastes by LC.

A liquid chromatographic method was developed for the simultaneous quantification of four major active components in tobacco (Nicotiana tobaccum L.) wastes. Samples were extracted with 70% v/v aqueous methanol, four compounds including chlorogenic acid, cryptochlorogenic acid, neochlorogenic acid and caffeic acid were identified and determined by using LC coupled to electrospray tandem mass spectrometry and LC-UV method, respectively. Separation in LC-UV was on an Alltima C18 column (250 mm × 4.6 mm i.d.; 5 µm) with a mobile phase consisting acetonitrile: ammonium acetate buffer (pH 4.5) (5:95 v/v), at a flow rate of 1.0 mL min-1, detected at 327 nm. Four regression equations showed good linear relationships (r 2 > 0.999) between the peak area of each marker and concentration. The method has good repeatability and precision, the intra-day and inter-day RSD for both retention time and peak area was less than 1.0%. The recoveries, measured at three concentration levels, varied from 96.33 to 101.10%. The LOD (S/N = 3) and LOQ (S/N = 6) were less than 0.010 and 0.795 µg·mL-1, respectively. This assay was successfully applied to the determination of four active compounds in ten samples. The results indicated that the developed assay method was rapid, accurate, reliable and could be readily utilized as a quantitative analysis method for various of tobacco wastes.

Caffeic acid (CA) protects cerebellar granule neurons (CGNs) from apoptosis induced by neurotoxin 1-methyl-4-phenylpyridnium (MPP~+) / 北京大学学报(医学版)

Objective: To assess the effects of caffeic acid (CA) on MPP + induced cerebellar granule neurons (CGNs) apoptosis. Methods: CGNs were pretreated with caffeic acid at 55, 110 and 220 ?mol/L for 6 h, then treated with 100 ?mol/L MPP + for 24 h (concentration effect relationship). In addition CGNs were pretreated with caffeic acid at 110 ?mol/L for 0 h, 6 h, 12 h, and 24 h, respectively, then treated with 100 ?mol/L MPP + for 24 h (time response relationship). Besides, after treatment with MPP + for 24 h, CGNs were incubated with caffeic acid at 55, 110 and 220 ?mol/L,respectively. Cell viability was determined by 3 (4,5 dimethylthiazol 2 yl) 2,5 diphenyltetrazolium bromide (MTT) assay and caspase 3 activity was assayed by caspase 3 fluorometric assay kit. Results: MTT assay revealed that caffeic acid significantly inhibited cell viability decrease induced by MPP +, and caspase 3 fluorometric assay showed that caffeic acid efficiently suppressed caspase 3 activation in CGNs induced by MPP +. Conclusion: Caffeic acid (CA) can significantly protect CGNs from apoptosis induced by MPP + and may provide a useful therapeutic strategy for the treatment of Parkinson's disease.

Inhibitory effect and induction of apoptosis of caffeic acid Ge on growth of U14 in mice / 中国病理生理杂志

AIM:To evaluate the antitumor effect of caffeic acid Ge on U14 tumor bearing mice.METHODS:The tumor inhibitory ratios of caffeic acid Ge on the growth of U14 in mice was observed.Apoptosis morphological transformation of U14 cells induced by caffeic acid Ge was detected by electronic scan microscope and MG-P staining.Alteration of cell cycle was analyzed by flow cytometry.Apoptosis-related protein levels of Bax and Bcl-2 were determined by immunity histochemistry technology.MTT assay was applied to study the antitumor activities of caffeic acid Ge in U14 cell lines in vitro.RESULTS:Tumor inhibitory rates in caffeic-acid Ge groups were 38.50%,47.17% and 64.02%(from low dose to high dose)(P