Spent hops extract (Humulus Lupulus L.) attenuates inflammation and angiogenesis of the retina via the nuclear factor-kappaB and protein kinase B/extracellular signal-regulated kinase pathways.

Spent hops extract (SHE) is a plant extract containing compounds with proven anti-inflammatory and anti-angiogenic activities. However, extract may exert synergic effects compared to its individual polyphenol components. Inflammatory diseases of the retina may lead to visual impairment, a reduction of the comfort of life, and even blindness due to the formation of new pathological blood vessels. More effective therapeutic options are being sought. The goal of the present study was to investigate the anti-inflammatory and anti-angiogenic potentials of SHE on human retinal pigment epithelial cells (ARPE-19) stimulated by lipopolysaccharide (LPS) or tumor necrosis factor alpha (TNF-α). The SHE (250 µg/mL) was found to downregulate the gene expression of interleukin 6 (IL-6) to 33% in LPS-triggered cells; it also reduced both matrix metalloproteinase 2 (MMP-2) and 9 (MMP-9) mRNA expression to 13% and 43% respectively, and their activity to 82% (MMP-2) and 57% (MMP-9), compared to TNF-α-stimulated cells. Also, SHE modulated the TNF-α-induced expression of vascular endothelial growth factor (VEGF) and endothelial growth factor receptor 2 (VEGFR2). It is possible that SHE inhibited retinal inflammation and angiogenesis by suppressing the nuclear factor kappa B (NF-κB), protein kinase B (Akt) and extracellular signal-regulated kinase (ERK) pathways. Our results demonstrate that SHE has anti-inflammatory and anti-angiogenic potential against retinal diseases. This is the first such study to report on the efficacy of SHE on retinal inflammatory diseases.

Chemopreventive properties of spent hops (Humulus Lupulus L.) extract against angiogenesis, invasion and migration of colorectal cancer cells.

There is a great deal of interest in identifying new chemopreventive agents for colorectal cancer, one of the leading causes of cancer-related death. One promising group of candidates is the polyphenols; being natural compounds with high structural diversity, they have a very wide spectrum of anti-inflammatory, anti-oxidant and anti-cancer properties. The present study reports for the first time that spent hops extract (SHE) inhibits the angiogenesis, invasion and migration of SW-480 and HT-29 colorectal adenocarcinoma cells; after incubation with 200 µg/mL SHE, SW-480 and HT-29 cell invasion fell by 98.5% and 89% vs. controls, and migration was inhibited by 99% and 88% vs. controls. These changes were accompanied by a decline of matrix metalloproteinase 2 (MMP-2) and MMP-9 expression and activity. In addition, SHE reduced the expression of vascular endothelial growth factor and hypoxia-inducible factor 1α for both cell lines, indicating that the tested extract has anti-angiogenic potential. In conclusion, our data shows that SHE may be an effective chemopreventive agent acting via the inhibition of angiogenesis, invasion and migration of colorectal cancer cells.

Evaluation of phenolic composition, antioxidant and cytotoxic activity of aronia melanocarpa leaf extracts.

The objective of this study was to investigate the phytochemical composition, antioxidant and cytotoxic potential of aronia leaf crude phenolic-extract (ACE) and purified phenolic-rich extract (APE) on human intestinal cells (CCD 841 CoN) and colon cancer cells (SW-480 and HT-29). UPLC-Q-TOF-MS analysis confirmed that aronia leaves are rich in structurally diverse polyphenols (25 and 42 compounds for ACE and APE, respectively). Chlorogenic acid and quercetin-3-rutinoside were most abundant in both aronia extracts. The sum of detected polyphenols varied significantly between extracts ranging from 32.8 mg/g (ACE) to 436.3 mg/g (APE). The biological potential of aronia extracts was confirmed by applying in vitro antioxidant and cytotoxic assays. The results of antioxidant activity (ABTS and FRAP) indicate that APE showed 2-fold stronger antioxidant properties compared to ACE. APE revealed a stronger cytotoxic effect on SW-480 and HT-29 cells than ACE (MTT test). After 48 -hours of incubation, APE was found to inhibit SW-480 cell growth by 50% vs. control at 194.35 µg/mL, while for HT-29 cells it was observed at 552.02 µg/mL. In the case of ACE, IC50 has not been reached for SW-480 cells after 48 -hours of treatment, but for HT-29 it was 794.84 µg/mL. Moreover, the viability was significantly decreased in a concentration- and time-dependent manner for both cancer cell lines. Examined extracts showed selective inhibitory potential against colon cancer cells. However, after 72 h incubation with CCD 841 CoN cells, the obtained IC50 values for APE and ACE were 594 µg/mL and 709 µg/mL respectively. This suggests that aronia leaves are valuable natural-based products that may support the treatment as chemopreventive agents in colorectal cancer.

Chemoprotective effects of Japanese quince (Chaenomeles japonica L.) phenol leaf extract on colon cancer cells through the modulation of extracellular signal-regulated kinases/AKT signaling pathway.

Japanese quince leaf phenol-rich extract (PRE) represents a good source of phenolic compounds, among which chlorogenic acid and naringenin hexoside are the main constituents. The aim of this research was to evaluate the chemopreventive activity of PRE in human colon cancer (SW-480 and HT-29) and human normal colon cell line (CCD 841 CoN). All cell lines were exposed to different concentrations of the extract (150-500 κg/mL for SW-480 and CCD 841 CoN; and 250-750 κg/mL for HT-29) to investigate migration and invasion, as well as the activity and secretion of metalloproteinases (MMP-2 and MMP-9) involved in these mechanisms. Moreover, the influence of PRE on the activity of ERK and AKT pathways, which are strongly involved in colon cancer development (CRC), were measured. Our results demonstrated that PRE significantly inhibited migration and invasion in SW-480, HT-29 and CCD 841 CoN cells through MMP-2 and MMP-9-dependent mechanisms. We also proved that PRE can effectively downregulate both the activity and protein expression of MMP-2 and MMP-9 in these cell lines. The exception was the higher concentration of PRE, which up-regulated the protein expression of MMP-9 in SW-480. Additionally, we showed that significant inhibition of p-ERK/p-AKT expression in SW-480 after treatment with PRE is involved in chemopreventive effects of this extract. In case of exposure of HT-29 cells to PRE, we observed a significant upregulation of p-ERK protein expression, and suppression of p-AKT mechanism. This research of Japanese quince phenol leaf extract suggests its application in colon cancer prevention and treatment due to its ability to inhibit migration and invasion in MMP-9 and MMP-2-dependent mechanisms via most likely the modulation of ERK and AKT signaling pathways in colon cancer cells. Overall, our results provide an experimental foundation for further research on its potential activities and effects in vivo.

The influence of polyphenol-rich extracts on the production of pro-inflammatory mediators in macrophages.

Recent decades have seen a rise in chronic inflammatory diseases such as diabetes, cardiovascular diseases, asthma, rheumatoid arthritis, neurodegenerative diseases. Importantly, such chronic inflammatory diseases also increase the risk of cancer development and there is a pressing need to identify new anti-inflammatory drugs. One promising source of new medication are natural polyphenolic compounds and polyphenol-rich preparations, extracts and foods, which have strong antioxidant properties. This paper reviews the anti-inflammatory role of polyphenolic-rich natural extracts, and their ability to modulate crucial pro-inflammatory mediators, such as cyclooxygenase-2, prostaglandin E2, inducible nitric oxide synthase, and nitric oxide, in macrophage cells. Our research confirms that natural compounds have health potential, and could be used in the treatment or prevention of inflammatory diseases.

Comparison of phenolic compounds, antioxidant and cytotoxic activity of extracts prepared from Japanese quince (Chaenomeles japonica L.) leaves.

Phenolic compounds are very important in the prevention and treatment of many civilization diseases, including colorectal cancer (CRC). In the present study we investigated and compared the phytochemical composition, antioxidant and cytotoxic activity of Japanese quince (Chaenomeles japonica L.) leaves crude phenolic extract (CPE) and purified phenolic-rich extracts (PRE). The UPLC-Q-TOF-MS analysis showed that both extracts contain diversified phenolics compounds (33 - 36 compounds in the PRE and CPE, respectively), among which chlorogenic acid and naringenin hexoside turned out to be the main constituents. Both FRAP and ABTS tests showed that PRE had 2-fold higher antioxidant activity compared to CPE. Furthermore, PRE exhibited a higher cytotoxic activity towards colon cancer cells (SW-480 and HT-29) than CPE. After 24-hours incubation with PRE the IC50 value for SW-480 cell line was obtained at the concentration of 239 µg/mL, while CPE treatment caused the same decrease only after 72h at 277 µg/mL. In addition, PRE had a stronger cytotoxic effect on the colon cancer cell lines (SW-480 and HT-29) than on normal intestinal cells (CCD-18Co and CCD 841 CoN). These results provide the first evidence that extracts from Japanese quince leaves (especially phenolic-rich extract, PRE) strongly decrease the viability of both SW-480 and HT-29 lines, which may suggest their cytotoxic activity towards colon cancer cells.

Spent hops (Humulus Lupulus L.) extract as modulator of the inflammatory response in lipopolysaccharide stimulated RAW 264.7 macrophages.

Macrophages play important roles in acute and chronic inflammation. Upon their activation, they secrete a variety of mediators, including eicosanoids, nitric oxide and cytokines, which play different roles in the stimulation and resolution of inflammatory processes. There is a continuous search for selective modulators of these processes. Natural polyphenols and polyphenol-rich extracts have been found to possess preventive and therapeutic potential, including by their anti-inflammatory effects. In this study, the inhibition of the formation of inflammatory mediators by the spent hops extract (SHE), a polyphenol-rich extract from Humulus Lupulus L., was examined using lipopolysaccharide (LPS)- activated murine macrophages (RAW 264.7). The SHE suppressed inter alia the interleukin-6 (IL-6) mRNA expression to 32% in LPS-activated macrophages and to 61% at a protein level (at 25 µg/mL). SHE reduced both the cyclooxygenase-2 (COX-2) mRNA expression to 47% and their protein expression to 32%. Not only did SHE inhibit the IL-6 and COX-2 levels but also decreased both inducible nitric oxide synthase (iNOS) protein expression to 2% at 25 µg/mL and nitric oxide (NO) production for all tested concentrations. The inhibited expression of these inflammatory molecules was likely caused by the reduced activity of nuclear factor-κB (NF-κB). Both mRNA and protein expression of NF-κB was decreased to 38% and 42%, respectively. These results provide the first evidence that SHE decreases the expression of proinflammatory cytokines and inflammatory mediators, which merits further studies to investigate the potential of SHE as anti-inflammatory preparation.

Japanese quince (Chaenomeles japonica) leaf phenol extract as modulator of the inflammatory response in lipopolysaccharide-triggered murine macrophage RAW 264.7 cells.

In the present study we investigated the anti-inflammatory activity of Japanese quince leaf polyphenol-rich extract (JQLPE) in lipopolysaccharide (LPS)-activated murine macrophages (RAW 264.7). The Q-PCR analysis revealed that JQLPE decreased Nfkb1, Ptgs2, and Il1b expression at the mRNA level by 80%, 50% and 48%, respectively. Similarly, JQLPE significantly attenuated expression of inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), interleukin-1beta (IL-1ß), IL-6 and tumor necrosis factor alpha (TNF-α) (by 60%, 50%, 67%, 37% and 36%, respectively) at the protein level and nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7. Western blot also showed that the expression of nuclear factor kappaB (NF-κB) p65 and p-NF-κB p65 was down-regulated after JQLPE treatment. These results provide the first evidence that JQLPE decreases the expression of pro-inflammatory cytokines (IL-1ß, IL-6, TNF-α), inflammatory mediators (COX-2, iNOS) and both NF-κB p65 and p-NF-κB p65 in LPS-stimulated RAW264.7 cells, which may suggest its anti-inflammatory activity.

Polyphenol extract from evening primrose pomace alleviates experimental colitis after intracolonic and oral administration in mice.

Oenothera paradoxa (EP) preparations are commonly used in folk medicine to treat skin diseases, neuralgia, and gastrointestinal (GI) disorders. Several reports suggested that EP preparations exhibit potent anti-inflammatory and antioxidant activities both in vitro and in vivo. Here, we aimed to characterize the action of EP pomace polyphenol extract in mouse model of colitis. We analyzed the composition of EP pomace polyphenol extract using reversed phase HPLC system and ultra-performance liquid chromatography (UPLC) system coupled with a quadrupole-time of flight (Q-TOF) MS instrument. Then, we used a well-established animal model of 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis to determine the anti-inflammatory action of EP pomace polyphenol extract. We also investigated the effect of the EP pomace polyphenol extract on pro-inflammatory (IL-1ß and TNF-α) cytokine mRNA levels and hydrogen peroxide concentration in the inflamed colon. Administration of EP pomace polyphenol extract significantly improved macroscopic and microscopic damage scores, as well as myeloperoxidase (MPO) activity in TNBS-treated mice. The anti-inflammatory effect of the extract was observed after intracolonic and oral administration and was dose-dependent. Significant reduction of tissue hydrogen peroxide level after treatment with EP pomace polyphenol extract suggests that its therapeutic effect is a result of free radical scavenging. This novel finding indicates that the application of the EP pomace polyphenol extract in patients with inflammatory bowel diseases (IBDs) may become an attractive supplementary treatment for conventional anti-inflammatory therapy.

WWOX, the tumour suppressor gene affected in multiple cancers.

WWOX is a tumour suppressor gene affected in multiple cancers, especially in breast, prostate and ovary. This gene is located at the chromosomal area 16q23.3-24.1, which was identified as a common chromosomal fragile site FRA16D. WWOX turned out to possess tumour suppressor features despite the fact that the most basic (classical) way of tumour suppressor gene inactivation involves both alleles (e.g. through deletions, point mutations and promoter methylation), which is very rare event in a case of WWOX, occurring only in few cell lines. A large number of papers corroborate the phenomenon of correlation between the loss of WWOX expression and more aggressive/worse prognosis in many different types of tumours, for example breast cancer, nonsmall cell lung cancer, bladder cancer, gastric cancer or sporadic meningiomas. Ectopically increased WWOX expression promotes migration through basal membrane, however suppresses anchorage independent growth and induces normal-like colony formation in matrigel.

The effect of interleukin-6 on hepatitis B virus replication in peripheral blood mononuclear cells in vitro.

Although the major target organ for hepatitis B virus (HBV) is the liver, the possibility of infection of peripheral blood mononuclear cells (PBMCs) with HBV has also been reported. This study was performed to analyze the course of HBV infection of PBMCs and to investigate the influence of interleukin-6 (IL-6) on the efficiency of infection of PBMCs with HBV in vitro. PBMCs isolated from a healthy donor were infected by exposing to a HBsAg-, HBeAg-positive serum in the presence or absence of exogenous IL-6. The efficiency of infection was estimated by HBV DNA determination in the cells and medium in the course of infection. The results of this study show that the presence of IL-6 during the PBMCs infection with HBV increased the efficiency of this infection.

[Evaluation of persistence of IFN-alpha treatment response in chronic hepatitis C patients according with HCV-RNA presence in PBMC]. / Ocena trwalosci odpowiedzi na leczenie IFN-alpha chorych na przewlekle zapalenie watroby typu C na podstawie obecnosci HCV-RNA w komórkach jednojadrowych krwi obwodowej.

BACKGROUND: The current criterion of cure of chronic hepatitis C (CHC) is the absence of HCV-RNA in hepatocytes and peripheral blood mononuclear cells (PBMC), as these cells are considered for the place of protracted viral replication. PURPOSE: Evaluation of HCV-RNA presence in serum, freshly isolated PBMC and cultured PBMC, taken from patients responding to interferon alfa2b therapy, after 42 months of follow-up. Fifty four adults with CHC was treated with interferon alfa-2b (IFN-alpha 2b). Evaluation of HCV-RNA presence in serum were performed using RT-PCR method, before the therapy, and after 6 and 18 months of follow-up. All patients received interferon alpha-2b for 6 months. After 18 months of follow-up, results of treatment (ALT activity and presence of HCV-RNA in serum) were retrospectively analyzed. In 10 of 54 patients (18.5%) normalization of ALT activity and negativization of HCV-RNA in serum were observed. After 42 months of follow-up, in these 10 patients, the presence of HCV-RNA in serum, in freshly isolated PBMC and cultured PBMC, were evaluated. RESULTS: After 42 months of follow-up, HCV-RNA was not found in serum of any patient. In 3 of 10 patients HCV-RNA was present in freshly isolated PBMC. After 3-day-culture of PBMC, HCV-RNA was found in another patient. After 6 days of culture, results of RT-PCR test were negative in all cases. CONCLUSIONS: Absence of HCV-RNA in serum as well as in freshly isolated PBMC after 42 months of follow-up after the treatment of CHC, is not synonymous with eradication of virus. Three-day-culture of PBMC may be a valuable method, allowing for detection of subtreshold amounts of HCV. Longer, 6-day-culture seems to be invaluable diagnostically.

Structure-activity relationship in a series of new 1-(2-pyrimidinyl)piperazine derivatives with hypnotic activity.

Preparation, biological properties and QSAR of new derivatives of 1-[4-(2-pyrimidinyl)-1-piperazinyl]-1, 3-butandione (11-13, and 15-18) and 3-[4-(2-pirimidinyl)-1-piperazinyl]-3-oxopropanoate (20-22) exhibiting hypnotic activity in mice are reported. The best therapeutic indices (TI = LD50/ED50) 4.7 and 9.4 for po and ip administration, respectively, were found for 1-[4-(2-pyrimidinyl)-1-piperazinyl]-2-n-pentyl-1,3-butandione (15). QSAR studies showed that the biological activity grows initially with an increase in lipophilicity to drop dramatically for log P > 2.5.