Małgorzata Kapral

Medical University of Silesia in Katowice, Catowice, Silesian Voivodeship, Poland

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Publications (34)24.27 Total impact

  • Malgorzata Kapral · Joanna Wawszczyk · S. Sośnicki · L. Wéglarz
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    ABSTRACT: Inflammatory bowel disease (IBD) is chronic inflammatory condition associated with increased risk of developing colorectal cancer. A number of mediators of inflammation, such as pro-inflammatory cytokines, prostaglandins and nitric oxide have been involved in carcinogenesis, especially in the promotion and progression stages. NO is synthesized from L-arginine by constitutively expressed endothelial and neuronal nitric oxide synthases (eNOS and nNOS, respectively) and an inducible NOS (iNOS) isoform expressed under inflammatory conditions. A selective inhibitors of iNOS could be, therefore, considered to be good candidates as chemopreventive agents against colon cancer. In this study, the effect of inositol hexaphosphate (IP6), dietary phytochemical, on the mRNA expression of iNOS stimulated with bacterial lipopolysaccharides (Escherichia coli and Salmonella typhimurium) and IL-1β in intestinal cells Caco-2 for 6 and 12 h was investigated. A transcription level of iNOS with the use real time QRT-PCR technique was determined in cells treated with 1 and 2.5 mM IP6. Stimulation of Caco-2 with pro-inflammatory factors (LPS and IL-1β) resulted in an up-expression of iNOS mRNA at 6 and 12 h. Cells exposed to IP6 only revealed significant reduction in iNOS gene transcription after 12 h. A decrease in iNOS transcription by IP6 following the gene induction by proinflammatory agents in 6 and 12 h lasting cultures was also determined. The findings of this study suggest that one of the anticancer and anti-inflammatory abilities of IP6 can be realized by suppressing the expression of gene encoding inducible nitric oxide synthase isoform at the transcriptional level.
    No preview · Article · Jul 2015 · Acta poloniae pharmaceutica

  • No preview · Article · Mar 2015
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    ABSTRACT: The aim of the present study was to examine the influence of IP6, a naturally occurring phytochem- ical, on the expression of genes coding for proliferation markers, i.e., cyclin D1 (CCND1) and histone H3 in IL-1β-stimulated intestinal cancer cell line Caco-2. Quantification of genes expression was carried out using real time RT-QPCR technique in Caco-2 cells after treatment with IL-1β, 1 and 2.5 mM of IP6 for 3, 6 and 12 h. In separate cultures, cells were incubated with IL-1β for the indicated times. The untreated Caco-2 cells were used as the control. In a time course experiment, stimulation of cells with IL-1β only resulted in an overex- pression of both CCND1 and histone H3 mRNAs as compared with control. IP6 had no influence on IL-1β-stimulated CCND1 expression for 3 and 6 h. After 12 h, statistically significant decrease in CCND1 mRNA was observed in cells exposed to IL-1β and IP6 (1 and 2.5 mM) in relation to cells treated with IL-1β only. The levels of H3 mRNA in IL-1β-stimulated cells and cells treated with IL-1β and IP6 revealed no statistically significant differences after 3 h. IP6 at 1 and 2.5 mM enhanced IL1β-stimulated transcription of H3 gene after 6 h. Subsequently (12 h), the combination of IP6 and IL-1β decreased H3 mRNA level compared to IL1β-treated cells. In conclusion, pro-inflammatory cytokine IL-1β up-regulates CCND1 and histone H3 mRNAs expression in Caco-2 cells. These results suggest that the ability of IP6 to inhibit colon cancer cells proliferation may be mediated through downregulation of genes encoding cyclin D1 and histone H3 at the mRNA level.
    No preview · Article · Nov 2014 · Acta poloniae pharmaceutica
  • Joanna Wawszczyk · Małgorzata Kapral · Andrzej Hollek · Ludmiła Węglarz
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    ABSTRACT: Colon cancer has been remaining the second leading cause of cancer mortality in Poland in the last years. Epidemiological, preclinical and clinical studies reveal that dietary phytochemicals may exert chemopreventive and therapeutic effect against colorectal cancer. There is a growing interest in identifying new biologically active agents from dietary sources in this respect. Pterostilbene (trans-3,5-dimethoxy-4-hydroxystilbene) is a naturally occurring stilbene, that has been found to have antioxidative, anti-inflammatory and antipro- liferative properties. Compared to other stilbenes, pterostilbene has greater bioavailability, and so, a greater potential for clinical applications. Recent studies showed that pterostilbene exhibits the hallmark characteristics of an anticancer agent. The aim of this study was to analyze antiproliferative and cytotoxic effects of pterostilbene on human colon cancer Caco-2 cells. They were cultured using standard techniques and exposed to increasing doses of pterostilbene (5-100 μM) for 48 and 72 h. Cell proliferation was determined by sulforhodamine B assay. The growth of treated cells was expressed as a percentage of that of untreated control cells. Pterostilbene decreased proliferation rate of Caco-2 cells in a dose- and time-dependent manner. Its concentrations = 25 μM did not affect cell growth after 48 h treatment period. Significant growth inhibition was observed in cultures incubated with higher concentrations of pterostilbene (40-100 μM). Pterostilbene at all concentrations used (5-100 μM) caused significant inhibition of cell proliferation when the experimental time period was elongated to 72 h. The maximum growth reduction was observed at 100 mM pterostilbene. The cytotoxicity of pterostilbene was evaluated in 48 h cultures based on lactate dehydrogenase (LDH) leakage into the culture medium and showed dose-related pattern. The findings of this study showed significant dose-dependent antiproliferative and cytotoxic effects of pterostilbene against human colon cancer cells in vitro.
    No preview · Article · Nov 2014 · Acta poloniae pharmaceutica
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    ABSTRACT: DNA microarray analysis through simultaneous measurement of expression of thousands of genes in the same tissue sample has enormous discovery potential. Comparison of gene expression profiles in disease with healthy tissues may highlight the involvement of both expected and unsuspected pathways leading to the formation of the pathology. Thus, genome wide expression array analysis has been successfully used to characterize a variety of different abnormal processes. Chronic rhinosinusitis (CRS) is a general term for multiple subsets of inflammatory conditions affecting paranasal cavities. Distinct clinical presentations and laboratory findings together with incomplete understanding of CRS pathophysiology make that disease an ideal candidate for genome wide expression array analysis. The potential development of the technique in CRS would be the definition of disease specific gene profiles enabling better understanding disease etiology. The purpose of this chapter is to describe DNA microarray contributions toward achieving a better understanding of CRS. The outcomes of DNA microarray analysis of sinus mucosa in patients with chronic rhinosinusitis with nasal polyps (CRSwNP) compared to healthy epithelium are described. We identified 556 genes, which were differentially expressed in CRS tissues compared to the control. Among them, 217 showed significantly higher expression, whereas 339 lower expression in CRSwNP than in the controls. In CRSwNP, significantly higher transcriptional activity of MMP10, NOS2A, ALOX15 and IL-8 genes was observed. In the control group, significantly higher expression of DMBT1, ALOX12 and LTF genes was detected. Those findings were confirmed via quantitative reverse transcription polymerase chain reaction (qRT-PCR). It might be concluded that the described technology can be successfully used to identify genes implicated in the pathogenesis of CRS. The results are discussed with the outcomes of other research in that field. The analysis of gene expression by microarray is a promising research method in the field of CRS. Some reports demonstrated that DNA microarray analysis is a feasible tool for studying different disease subclassifications in CRS. Moreover, molecular alterations obtained using that technique may indicate changes during the clinical course of CRS. Defining gene expression profiles with DNA microarray may help elucidate new prognostic factors and support development of new therapeutic alternatives.
    No preview · Chapter · Jan 2014
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    ABSTRACT: Transforming growth factor β (TGF- β ) is a multifunctional cytokine recognized as an important regulator of inflammatory responses. The effect of inositol hexaphosphate (IP6), a naturally occurring phytochemical, on the mRNA expression of TGF- β 1, TGF- β 2, TGF- β 3 and T β RI, T β RII, and T β RIII receptors stimulated with bacterial lipopolysaccharides (Escherichia coli and Salmonella typhimurium) and IL-1 β in intestinal cells Caco-2 for 3 and 12 h was investigated. Real-time qRT-PCR was used to validate mRNAs level of examined genes. Bacterial endotoxin promoted differential expression of TGF- β s and their receptors in a time-dependent manner. IL-1 β upregulated mRNA levels of all TGF- β s and receptors at both 3 h and 12 h. IP6 elicited the opposed to LPS effect by increasing downregulated transcription of the examined genes and suppressing the expression of TGF- β 1 at 12 h. IP6 counteracted the stimulatory effect of IL-1 β on TGF- β 1 and receptors expression by decreasing their mRNA levels. IP6 enhanced LPS- and IL-1 β -stimulated mRNA expression of TGF- β 2 and - β 3. Based on these studies it may be concluded that IP6 present in the intestinal milieu may exert immunoregulatory effects and chemopreventive activity on colonic epithelium under inflammatory conditions or during microbe-induced infection/inflammation by modulating the expression of genes encoding TGF- β s and their receptors at transcriptional level.
    Preview · Article · Dec 2013 · Mediators of Inflammation
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    ABSTRACT: Phytic acid (IP6) is a major fiber-associated component of a diet physiologically present in human intestines. Studies showed that this phytochemical can modulate immune functions of intestinal epithelium through regulation of proinflammatory cytokines secretion but mechanisms underlying these cellular response to IP6 have weakly been examined, as yet. The aim of this study was to determine the role of protein tyrosine kinase (PTK) in secretion of IL-8, a central proinflammatory cytokine, by unstimulated and IL-1beta-stimulated intestinal epithelial cells Caco-2 treated with IP6 (1 and 2.5 mM). To study the involvement of PTK signal pathway in IL-8 secretion, inhibitors of phosphotyrosine phosphatase (sodium orthovanadate, OV) and tyrosine kinase (genistein, GEN) were incubated with Caco-2 cells prior to IP6 treatment. IP6 had suppressive effect on basal and IL-1beta-stimulated IL-8 secretion by cells. The effect of OV on IL-8 release by cells treated with IP6 was different under constitutive and stimulated conditions. Secretion of IL-8 was significantly down-regulated in cells with GEN and GEN plus IP6 treatment. In addition, total PTK activity in both unstimulated and IL-1beta stimulated cells was determined in the presence of IP6. The results suggest that physiological intestinal concentrations of IP6 may have an inhibitory effect on IL-8 secretion by Caco-2 cells and one of the mechanisms of its action is the inhibition of PTK signaling cascade. The study revealed for the first time that PTKs could be one of the molecular targets for IP6 effects in the intestinal epithelial cells.
    No preview · Article · Apr 2013 · Acta poloniae pharmaceutica
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    ABSTRACT: BACKGROUND: The inflammatory process underlying nasal polyposis is induced and perpetuated by the enhanced activity of several agents including transcription factors. It has recently been demonstrated that one of them, named nuclear factor-kappa B (NF-κB), is implicated in the regulation of multiple pro-inflammatory genes. OBJECTIVES: The aim of the study was to identify using microarray technology which NF-κB-dependent genes are activated in nasal polyp (NP) samples compared to the control mucosa. MATERIAL AND METHODS: The transcriptional activity of genes was analyzed using an oligonucleotide microarray on 15 NPs and 8 cases of normal nasal mucosa. RESULTS: Gene expression patterns obtained in NPs were significantly different from those in normal mucosa. NPs and control cases clustered separately, each of them with large homogeneity in gene expression. Among 582 human NF-κB-dependent genes 25 showed a significantly higher expression in NPs compared to the control. The largest increase focused on gene encoding TFF3 (a 5-fold higher expression) followed by NOS2A (5x), SERPINA1 (4x), UCP2 (4x), OXTR (4x) and IL8 (3x) (p<0.05). In healthy mucosa 19 genes presented increased transcription activity compared to NPs. The most significantly enhanced levels were shown LTF gene (20 fold) followed by KRT6B (7x), LYZ (7x), SD11B2 (5x) and MMP3 (4x) (p<0.05). CONCLUSIONS: DNA microarray technology highlights the involvement of many unsuspected pathologic pathways which could be involved in NP growth. The identification of novel disease-related genes may help to understand the biology of NPs and elaborate new targeted therapy.
    Full-text · Article · Mar 2013 · Advances in Clinical and Experimental Medicine
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    ABSTRACT: Inositol hexaphosphate (IP6) is a naturally occurring phytochemical, found in abundance in cereals, legumes and other high-fiber-content diets. IP6 has shown promising efficacy against a wide range of cancers. Its anti-cancer activity involves anti-proliferative, pro-apoptotic and anti-metastatic effects. Both matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs), are implicated in tumor growth, metastasis, and angiogenesis. Phorbol-12-myristate 13-acetate (PMA) is a well-known inflammatory stimulator and tumor promoter that activates PKC and increases the invasiveness of various types of cancer cells by activating MMPs. The aim of the present study was to examine the influence of IP6 on the expression of selected MMPs, i.e., MMP-1, -2, -3, -9, 10, -13 and their TIMP-1 and -2 in unstimulated and PMA-stimulated colon cancer cell line Caco-2. Quantification of genes expression in Caco-2 cells treated with 100 ng/mL of PMA, 2.5 mM of IP6 and both for 6 and 12 h was carried out using real time QRT-PCR technique. Stimulation of cells with PMA resulted in an up-expression of MMP-2, MMP-3, MMP-9, MMP-10, MMP-13 and TIMP-1 mRNAs and decrease in MMP-1 gene expression. The quantity of TIMP-2 transcript was reduced by PMA. A significant decrease in MMP-2, MMP-3, MMP-10, MMP-13, and TIMP-1 expression in response to IP6 was observed. IP6 down-regulated MMP-9 transcription induced by PMA and decreased the level of both MMP-2 and MMP-3 mRNAs in PMA-stimulated cells. Caco-2 treated with both PMA and IP6 showed a significant decrease in MMP-1 expression in comparison to PMA-stimulated cells. The results of this study show that PMA can modulate MMP and TIMP genes transcription in colon cancer cells Caco-2. IP6 exerts an influence of basal mRNA expression of some MMPs and their tissue inhibitors and down-regulates MMP-1, MMP-2, MMP-3 and MMP-9 in cells treated with PMA. IP6 could be an effective anti-metastatic agent that suppresses expression of MMP genes at transcription level.
    No preview · Article · Jan 2013 · Acta poloniae pharmaceutica
  • Małgorzata Kapral · Joanna Wawszczyk · Andrzej Hollek · Ludmiła Weglarz
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    ABSTRACT: Transforming growth factors-beta (TGF-beta) are multifunctional cytokines involved in the regulation of cell development, differentiation, survival and apoptosis. They are also potent anticancer agents that inhibit uncontrolled proliferation of cells. Incorrect TGF-beta regulation has been implicated in the pathogenesis of many diseases including inflammation and cancer. In humans, the TGF-beta family consists of three members (TGF-beta1, 2, 3) that show high similarity and homology. TGF-betas exert biological activities on various cell types including neoplastic cells via their specific receptors. Inositol hexaphosphate (phytic acid, IP6), a phytochemical has been reported to possess various health benefits. The aim of this study was to examine the effect of IP6 on the expression of genes encoding TGF-beta1, TGF-beta2, TGF-beta3 isoforms and their receptors TbetaRI, TbetaRII, TbetaRIII in human colorectal cancer cell line Caco-2. The cells were treated with 0.5, 1 and 2.5 mM IP6 for 3, 6 and 12 h. The untreated Caco-2 cells were used as the control. Quantification of genes expression was performed by real time QRT-PCR technique with a SYBR Green I chemistry. The experimental data revealed that the TGF-beta1 mRNA was the predominant isoform in Caco-2 cells and that IP6 enhanced transcriptional activity of genes of all three TGF-beta isoforms and their receptors TbetaRI, TbetaRII TbetaRIII in these cells. At concentrations up to 1 mM, IP6 over-expressed the genes in 6 h lasting cultures, and its higher dose (2.5 mM) caused successively increasing transcript level of TGF-beta isoforms and receptors with the duration of experiment up to 12 h. The findings of this study indicate that one of anti-cancer abilities of IP6 can be realized by enhancing the gene expression of TGF-beta isoforms and their receptors at the transcriptional level.
    No preview · Article · Nov 2012 · Acta poloniae pharmaceutica
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    ABSTRACT: Phytic acid (IP6) is an essential component of high fiber diet physiologically present in human large gut. It has been recognized to possess various significant health benefits effects including chemopreventive and have antineoplastic activity against various types of cancer. Moreover, its role in immune response through modulation of the secretion of proinflammatory cytokines and chemokines has been postulated. One of the signal transduction pathways involved in a variety of inflammatory responses is p38 mitogen-activated protein kinase (MAPK) pathway. The aim of this study was to examine effect of IP6 on human p38alpha MAP kinase activity and the expression of gene encoding p38 MAP kinase in unstimulated and IL-1beta-stimulated Caco-2 cells. Furthermore, the role of signaling pathways involving p38 MAP kinase in IP6-induced down-regulation of IL-8 secretion by unstimulated and IL-1beta-stimulated cells in the presence of p38 MAP kinase activator (anisomycin) and inhibitor (SB 203580) was evaluated. IP6 inhibited activity of recombinant p38 MAPK activity in dose-dependent manner. Treatment of cells with IP6 for 3 h resulted in decreased p38 MAP kinase expression in both unstimulated and stimulated with IL-1beta cells. The similar level of p38alpha mRNA was found in untreated and treated with IP6 cells after 6 and 12 h. Incubation of Caco-2 cells with anisomycin resulted in upregulation of IL-8 secretion and their pretreatment with anisomycin prior to IP6 addition showed down-regulation of IL-8 secretion compared to cells treated with anisomycin alone. The findings of this study show that p38 MAPK could be one of the molecular targets for IP6 in the intestinal epithelial cells and that IP6 inhibitory effect on IL-8 secretion by Caco-2 cells could be mediated by its inhibition of p38 activity.
    No preview · Article · Nov 2012 · Acta poloniae pharmaceutica
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    ABSTRACT: The latest studies suggest that adhesion molecules are involved in the arising of malignant changes and in distant metastasis induction. The soluble forms of several adhesion molecules, have recently emerged as novel and potentially useful tumor markers. Among a number of identified, high interest wake soluble molecules similar to the immunoglobulin -- soluble intercellular adhesion molecules-1 (sICAM-1) and soluble E-cadherin (sE-cadherin). In the present work, the authors concentrate on one tumor type, colorectal carcinoma, in which distant metastases, are the main cause of failure, in spite of surgical curing of the primary tumor. It is known that TNF-alpha (tumor necrosis factor - alpha) serum concentration of patients with cancer is raised. The changes in soluble adhesion molecules concentrations in serum and others fluids, could be modulated by many different factors affecting cancer cells. In the case of colon cancer one of the factors is a high-fiber diet, containing an anti-cancer chemical, inositol hexaphosphate (IP6). The aim of this study was to estimate the influence of TNF-alpha on the concentration of sICAM-1 and sE-cadherin in the microenvironment of HT-29 malignant epithelial colorectal cells stimulated with IP6. Additonally, adhesive property of HT-29 human colorectal cancer cell line to collagen I was estimated. The HT-29 cells were treated with TNF-alpha (10 ng/mL and 100 ng/mL - estimation of sICAM and sE-cadherin concentration; 100 ng/mL - adhesion assay), IP6 (0.5 mM, 1.0 mM, 2.0 mM) and TNF-alpha in combination with IP6. The level of sICAM-1 and sE-cadherin in cultures of HT-29 cells was measured by enzyme-linked immunosorbent assay (R&D Systems), and adhesion of the cells to collagen I was investigated by Cyquant Proliferation Assay Kit. The present findings demonstrate that TNF-alpha and inositol hexaphosphate have an effect on the sICAM-1 and sE-cadherin concentration in cultures of HT-29 cells. IP6 at a concentration of 2.0 mM induced a decrease of sE-cadherin concentration in cultures of these cells and significantly reduced their adhesion to collagen I. TNF-alpha at concentration of 100 ng/mL caused the significant increase in the sICAM-1 level, but to a lesser degree in the presence of higher concentrations of IP6. However, TNF-alpha did not cause such a significant increase in sE-cadherin level. The sE-cadherin concentration was most likely associated with inositol hexaphosphate activity.
    No preview · Article · Nov 2012 · Acta poloniae pharmaceutica
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    ABSTRACT: Matrix metalloproteinases (MMPs) have repeatedly been shown to play a very active role in extracellular matrix degradation associated with tumor invasion and metastasis. Tissue inhibitors of MMPs (TIMPs) are well-known for their ability to inhibit MMP activity thereby inhibiting malignant progression. Inositol hexaphosphate (IP6 phytic acid) has been recognized to have both preventive and therapeutic effects against various cancers including that of colon. In in vitro studies, IP6 has been demonstrated to inhibit cancer cell adhesion and migration. In the present study, the effect of IP6 on the expression of MMP and TIMP genes was evaluated in unstimulated and IL-1β-stimulated colon cancer cell line Caco-2. Real-time QRT-PCR was used to validate the transcription level of selected MMP and TIMP genes in Caco-2 cells after treatment with 1 ng/ml of IL-1β, 2.5 mM of IP6, and both for 6, 12, and 24 h. Stimulation of cells with IL-1β only resulted in an overexpression of MMP and their TIMP mRNAs. A significant decrease in MMP-13, MMP-3, MMP-2, and TIMP-1 basal expression was achieved by IP6. IP6 was also an efficient downregulator of MMP-1, MMP-9, and TIMP-2 genes transcription stimulated by IL-1β in 6 h lasting culture. After 12 h, IL-1β-induced MMP-2 mRNA expression was significantly reduced by IP6. Proinflammatory cytokine IL-1β upregulates MMP and TIMP mRNAs expression in colon cancer epithelial cells Caco-2. IP6 (2.5 mM) influences constitutive expression of both MMP and TIMP genes and downregulates IL-1β stimulated transcription of some of these genes. IP6 exerts its anti-metastatic activity through modulation of MMP and TIMP genes expression to prevent cancer cell migration and invasion.
    Full-text · Article · Mar 2012 · International Journal of Colorectal Disease
  • Joanna Wawszczyk · Małgorzata Kapral · Andrzej Hollek · Ludmiła Weglarz
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    ABSTRACT: Intestinal epithelial cells play an important role in the mucosal immune and inflammatory reactions via the expression and secretion of proinflammatory cytokines such as interleukin-6 (IL-6) and interleukin-8 (IL-8). The expression of both interleukins is regulated by nuclear factor KB (NF-kappaB). Phytic acid (IP6) is an essential component of high fiber diet. It is physiologically present in the human large gut at concentrations reaching 4 mM. It exhibits pleiotropic health beneficial effects including anti-oxidant and anti-tumor activities. Recent studies showed that IP6 can modulate immune functions of intestinal epithelium through regulation of proinflammatory cytokines secretion. The aim of this study was to analyze the effect of IP6 on the expression of IL-6 and IL-8 as well as p50 and p65 subunits of NF-kappaB and its inhibitor IkappaBalpha in Caco-2 cells stimulated with IL-1beta. A kinetic study of mRNAs expression in cells was performed after their treatment with 1 and 2.5 mM IP6 for 3, 6 and 12 h. Quantification of the genes expression was carried out using real time QRT-PCR technique. IP6 at all used concentrations had no influence on transcription of p65 gene and modulated expression of p50 and IkappaBalpha genes in Caco-2 cells. Treatment of cells with IP6 resulted in a marked decrease in both IL-6 (at 3 and 6 h) and IL-8 expression (3 h). The results of these studies suggest that IP6 may exert immunoregulatory effects on intestinal epithelium by influencing transcriptional activity of genes encoding p50 subunit of NF-kappaB, its inhibitor IkappaBalpha and proinflammatory cytokines IL-6 and IL-8.
    No preview · Article · Nov 2011 · Acta poloniae pharmaceutica
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    ABSTRACT: Matrix metalloproteinases 2 (MMP-2) and 9 (MMP-9) belong to a zinc dependent family of enzymes that degrade components of extracellular matrix. One postulated mechanism by which inositol hexaphosphate (phytic acid, IP6), an ubiquitous plant component, prevents the activation of MMPs may be due to its ability to chelate minerals. The aim of the study was to evaluate the expression profile of MMP-2, MMP-9 and their tissue inhibitors TIMP-1 and TIMP-2 at the mRNA level in human colorectal cancer cell line Caco-2 treated with IP6. A kinetic study of MMP-2, MMP-9 and TIMP-1, TIMP-2 mRNAs was performed after cells treatment with 1; 2.5; 5 mM IP6 for 1, 6, 12 and 24 h. Quantification of genes expression was carried out using real time QRT-PCR technique. The gene encoding MMP-9 was neither constitutively expressed nor induced by IP6 in Caco-2 cells. IP6 at the concentration of 1 mM evoked increase in MMP-2 transcript level, however, its higher doses (2.5; 5 mM) caused a decrease in this gene expression at 1 h incubation. In 24 h lasting culture along with increasing IP6 concentration, the cells expressed lower and lower MMP-2 mRNA level. In response to 1 and 2.5 mM at 6 h, the cells demonstrated an increased transcriptional activity of the TIMP-2 gene which was accompanied by a decrease in TIMP-1 gene transcription. Treatment of cells with 2.5 mM IP6 at 12 h resulted in a strong increase in both TIMP-1 and TIMP-2 expression. The results of this study show that IP6 modulates MMP-2, TIMP-1 and TIMP-2 genes expression in colon cancer cells at the transcriptional level in a way dependent on its concentration and time of interaction.
    Preview · Article · Nov 2010 · Acta poloniae pharmaceutica
  • Małgorzata Kapral · Joanna Wawszczyk · Sławomir Smolik · Ludmiła Weglarz

    No preview · Article · Nov 2010 · Acta poloniae pharmaceutica
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    ABSTRACT: Familial hypertrophic cardiomyopathy (FHCM) is characterized by an autosomal dominant transmission, left ventricular hypertrophy and myocardial disorganization. So far, 13 genetic loci and more than 130 mutations in ten different genes have been identified. Recent study suggested impaired force production associated with inefficient use of ATP as the main disease mechanism. We performed haplotype analysis with the use of microsatellite markers linked with beta-myosin heavy chain, troponin T, alpha-tropomyosin and cardiac myosin protein C genes in three Polish families with hypertrophic cardiomyopathy (23 individuals). This method is based on the analysis of distribution of the disease in the family and the alleles of chosen microsatellite markers. In two families, the disease was associated with beta-myosin heavy chain gene. We also found a genetic carrier of the mutated gene among children of the patients. In one family the connection of the disease with the mutation in alpha-tropomyosin gene was confirmed, no sudden cardiac deaths were recorded and the degree of myocardial hypertrophy was small.
    No preview · Article · Nov 2010 · Acta poloniae pharmaceutica
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    ABSTRACT: Quantification of p65, p50 and IκBα mRNAs was performed by real time QRT-PCR in Caco-2 cells treated with 10, 50, and 100 μg/mL of Desulfovibrio desulfuricans LPS for 1, 6, 12, and 24 h. A strong increase in expression of p65 and IλBα genes was induced by 10 and 100 μg/mL of LPS at 1 h; after 6 h higher transcript amounts of both genes were observed at 100 μg/mL LPS. The p65 expression level was significantly increased by 50 and 100 μg/mL at 12 h and lowered by all LPS doses at 24 h. No significant differences between IκBα mRNA quantity in cells exposed to LPS at 12 and 24 h were observed. No changes in expression of p50 mRNA were induced by LPS. The expression of p65 gene positively correlated with IκBα gene expression. D. desulfuricans LPS is capable of modulating transcriptional activity of p65 and IκBα genes in intestinal epithelial cells.
    No preview · Article · Nov 2010 · Folia Microbiologica
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    ABSTRACT: Matrix metalloproteinases (MMP) represent the family of endopeptidases that digest components of the extracellular matrix, including basement membranes. Specific tissue inhibitors (TIMP) control metalloproteinases activity. Breakage of matrix barriers enables tumor cells to invade the surrounding tissues and lymph nodes. The aim of the study was to investigate the mRNAs expression of MMP1, MMP2, MMP3, MMP14, and their tissue inhibitors in laryngeal squamous cell carcinoma and adjacent nonneoplastic tissues. Comparison between the mRNA levels of MMPs and TIMPs in tumor samples with lymph node metastasis and without metastatic lymph nodes was made as well. The study included 32 patients with primary laryngeal squamous cell carcinoma. Quantification of genes expression was performed by real time QRT-PCR technique. Expression of all examined MMPs (p<0,01) and TIMPs (p<0,05) was significantly higher in tumor areas. Comparative analysis of all MMP transcripts both in tumor and normal tissues showed the lowest transcriptional activity for MMP1 gene (p<0,01). In tumor and surrounding tissues MMP2, MMP3 and MMP14 genes revealed insignificant difference of transcriptional activity. Expression of TIMP2 mRNA compared with TIMP1 mRNA was higher in tumors (p=0,015), although in adjacent nonneoplastic tissues both mRNAs manifested the same level (p=0,111). Additionally, strong positive correlation between expression of MMP and their tissue inhibitors encoding genes in tumor center (R>0,5; p<0,05) was found. The changes in transcriptional activity of matrix metalloproteinases and their tissue inhibitors may be associated with tumor progression, invasion and metastasis.
    No preview · Article · Jan 2010 · Farmaceutyczny Przeglad Naukowy
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    ABSTRACT: Lipopolysaccharide (LPS, endotoxin) is a component of Gram-negative bacteria outer membrane. This, per se non-toxic, lipid-carbohydrate heteropolymer, when liberated from cell during its lysis, can trigger host immune system response. The intensity of this response depend on concentration and the chemical structure of LPS. Plasma proteins (lipoproteins, LBP - lipopolysaccharide-binding protein) bind endotoxin neutralizing it and facilitating its interaction with receptors (CD14, TLR4, CD11/CD18) on the surface of monocytes, macrophages, blood platelets and endothelial cells. Interaction of LPS with receptors activates the signaling pathway leading to liberation of inflammatory mediators such interleukins (IL-1, IL-6, IL-8, IL-12), tumor necrosis factor (TNF-α), products of arachidonic acid metabolism (leukotrienes and prostaglandins), nitric oxide and oxygen free radicals. These compounds, liberated by LPS stimulated immune system cells are pyrogenic, bactericidal and chemotactical. They also activate the complement and blood coagulation cascade. Inflammatory mediators aggravate inflammation enabling macroorganism to fight infection. However, the high concentration of endotoxin in bloodstream causes excessive secretion of these mediators leading to homeostasis disturbance and septic shock.
    No preview · Article · Jan 2010 · Farmaceutyczny Przeglad Naukowy