Expression of ABCA1 Transporter and LXRA/LXRB Receptors in Placenta of Women with Late Onset Preeclampsia.

BACKGROUND: Appropriate levels of cholesterol are necessary for the mother and developing fetus, but theirexcess may cause preeclampsia. The ABCA1 transporter mediates the secretion of cholesterol and is highly regulated at the transcriptional level via the nuclear liver X receptors (LXRs). METHODS: Sixteen preeclamptic and 39 normotensives healthy women with uncomplicated pregnancies were involved in the case-control study. The placental levels of ABCA1, LXRA and LXRB mRNA were quantified by real-time quantitative PCR. The concentrations of ABCA1, LXRA and LXRB proteins from the placenta were determined using an enzyme-linked immunosorbent assay Results: We found in the logistic regression model significantly lower placental expression of LXRB mRNA (crude OR = 0.26, 95% CI: 0.07-0.94, p = 0.040) and LXRA protein level (crude OR = 0.19, 95% CI: 0.05-0.69, p = 0.012) in late-onset preeclamptic women compared to healthy pregnant women. The values remained statistically significant after adjustment for possible confounders. CONCLUSIONS: Our results suggest that high placenta LXRA mRNA and LXRA protein expression levels decrease the risk of late-onset preeclampsia. These nuclear receptors could play a role in the development of preeclampsia through disturbances of lipid metabolism.

Combined Effects of Methyldopa and Flavonoids on the Expression of Selected Factors Related to Inflammatory Processes and Vascular Diseases in Human Placenta Cells-An In Vitro Study.

The aim of the study was to investigate combined effects of flavonoids (apigenin, baicalein, chrysin, quercetin, and scutellarin) and methyldopa on the expression of selected proinflammatory and vascular factors in vitro for prediction of their action in pregnancy-induced hypertension. The research was conducted on a trophoblast-derived human choriocarcinoma cell line and a primary human umbilical vein endothelial cell line. Cytotoxicity of compounds in selected concentrations (20, 40, and 100 µmol) was measured using the MTT test and the concentration of 40 µmol was selected for further analysis. Subsequently, their effects with methyldopa on the expression of selected markers responsible for inflammation (TNF-α; IL-1ß; IL-6) and vascular effects (hypoxia-inducible factor 1α-HIF-1α; placental growth factor-PIGF; transforming growth factor ß-TGF-ß; vascular endothelial growth factor-VEGF) at the mRNA and protein levels were assessed. It was found that every combined administration of a flavonoid and methyldopa in these cells induced a down-regulating effect on all tested factors, except PIGF, especially at the mRNA expression level. As hypertension generally raises TNF-α, IL-1ß, IL-6, HIF-1α, TGF-ß, and VEGF mRNA expression and/or protein levels, the results obtained in the studied model may provide a positive prognostic factor for such activity in vivo.

Theoretical Investigations on Interactions of Arylsulphonyl Indazole Derivatives as Potential Ligands of VEGFR2 Kinase.

Vascular endothelial growth factor receptor 2 (VEGFR2) is a key receptor in the angiogenesis process. The VEGFR2 expression is upregulated in many cancers so this receptor is an important target for anticancer agents. In the present paper, we analyse interactions of several dimeric indazoles, previously investigated for anticancer activity, with the amino acids present in the VEGFR2 binding pocket. Using the docking method and MD simulations as well as theoretical computations (SAPT0, PIEDA, semi-empirical PM7), we confirmed that these azoles can efficiently bind into the kinase pocket and their poses can be stabilised by the formation of hydrogen bonds, π-π stacking, π-cation, and hybrid interactions with some amino acids of the kinase cavity like Ala866, Lys868, Glu885, Thr916, Glu917, and Phe918.

The influence of certain plant substances and their chemopreventive activity in ovarian cancer.

A steadily growing number of studies have confirmed the beneficial effects of plant-derived substances (preparations) on the effectiveness of pharmacotherapy for ovarian cancer. A prior or parallel application of plant-derived substances and chemotherapy could be the way to strengthen the classic pharmacological treatment. Our paper presents several plant-derived substances with proven antiproliferative activities, in which phenolic and flavonoid bioactive compounds dominate, with particular emphasis on ovarian cancer cells. We are of the opinion that our paper will contribute to better understanding of the molecular basis for the positive interaction effect of concomitant application of the abovementioned plant substances with certain cytostatics. Also, this work may increase the number of preclinical in vivo experiments using these and other phenolic, flavonoid-rich plant substances to better understand their efficacy and safety and, in the future, to initiate clinical trials in this field.

Screening for impact of popular herbs improving mental abilities on the transcriptional level of brain transporters.

There are a number of compounds that can modify the activity of ABC (ATP-binding cassette) and SLC (solute carrier) transporters in the blood-brain barrier (BBB). The aim of this study was to investigate the effect of natural and synthetic substances on the expression level of genes encoding transporters present in the BBB (mdr1a, mdr1b, mrp1, mrp2, oatp1a4, oatp1a5 and oatp1c1). Our results showed that verapamil caused the greatest reduction in the mRNA level while other synthetic (piracetam, phenobarbital) and natural (codeine, cyclosporine A, quercetin) substances showed a selective inhibitory effect. Further, the extract from the roots of Panax ginseng C. A. Meyer exhibited a decrease of transcription against selected transporters whereas the extract from Ginkgo biloba L. leaves resulted in an increase of the expression level of tested genes, except for mrp2. Extract from the aerial parts of Hypericum perforatum L. was the only one to cause an increased mRNA level for mdr1 and oatp1c1. These findings suggest that herbs can play an important role in overcoming the BBB and multidrug resistance to pharmacotherapy of brain cancer and mental disorders, based on the activity of selected drug-metabolizing enzymes and transporters located in the BBB.

Effect of Epilobium angustifolium and Serenoa repens extracts on regulation of non-genomic signaling pathway of kinases.

OBJECTIVES: Changes of kinase activity of non-genomic cellular signaling pathway may influence the effectiveness of pharmacotherapy in case of hormone-dependent tumors. Our study investigated a possible interaction at the molecular level between an aqueous herbal extract of Epilobium angustifolium as well as a lipid-sterolic fruit extract of Serenoa repens and synthetic drugs used in the treatment of hormone-dependent cancers. MATERIAL AND METHODS: E. angustifolium and Serenoa repens extracts were orally administered to testosterone-induced rats for 21 days. Changes of RafA/Mapk3/Mapk1 mRNA levels were analyzed by real-time quantitative PCR using target specific primers. RESULTS: The level of RafA mRNA slightly increased in rats receiving Epilobium angustifolium (p = 0.076) and Serenoa repens (p = 0.016) extracts. Administration of these extracts resulted in significantly elevated Mapk1 and Mapk3 transcripts in the investigated animals (p < 0.05 for each extract). The levels of Mapk1 and Mapk3 mRNA strongly increased (p < 0.05 for each extract) in animals receiving concomitantly testosterone and the extracts, while RafA transcription slightly decreased (p < 0.05), as compared to controls. CONCLUSIONS: The results of our study may indicate a potential effect of S. repens and E. angustifolium extracts on the functioning of non-genomic cellular signaling kinases pathway. We investigated safety of these extracts to detect possible drug interactions between synthetic drugs used in the treatment of proliferative changes in hormone-dependent reproductive organs and herbal preparations.

[Modern diagnostics of osteoporosis based on the use of biochemical markers of bone turnover]. / Nowoczesna diagnostyka osteoporozy w oparciu o wykorzystanie biochemicznych markerów obrotu kostnego.

Osteoporosis is a disease with low bone mass and disorganization of the internal microarchitecture of bone tissue. Determination of biochemical markers allows for early diagnosis of changes in bone tissue metabolism. The search for a marker whose biological function could be directly connected with bone metabolism, clearly indicating a connection between its concentration and risk fracture as well as response to treatment, continues. Currently measurement of collagen-derived markers of bone resorption is used in the majority of cases. They are, first of all, telopeptides of collagen type 1 localized on the amino end-N-terminal telopeptide of type 1 collagen (NTX), as well as on the carboxy end-C-telopeptide of type 1 collagen (CTX) of collagen molecule. Among markers of bone synthesis, special attention is paid to the procollagen type 1 carboxy-terminal propeptide (POCP) and procollagen type 1 amino-terminal propeptide (P1NP). Simultaneous application of bone synthesis and resorption markers allows for a full imaging of the bone remodeling process and application of biochemical markers in the diagnosis and therapy of osteoporosis.

The influence of a standardized soybean extract (Glycine max) on the expression level of cytochrome P450 genes in vivo.

OBJECTIVE: Soybean isoflavones are phytoestrogens that reduce menopausal symptoms and decrease the risk of certain chronic diseases, such as cancer and cardiovascular diseases. Despite the widespread use of soybean isoflavones as functional food and dietary supplements, data regarding the safety as well as herb-drug interactions, remain scarce. The aim of the study was to investigate the influence of soybean extract on the expression levels of cytochrome P450 (CYP) genes using real-time PCR (RT-PCR). MATERIALS AND METHODS: Male Wistar rats were fed a standardized soybean extract containing 37% isoflavones (100 mg/kg) for 3 and 10 days. cDNA was synthesized from total RNA isolated from the liver using reverse transcription. The level of CYP genes expression was analyzed using RT-PCR method. RESULTS: Soybean extract administration resulted in a significant increase of CYP1A1 expression level compared with the control group (1.5-fold; p < 0.05). An inductory effect was also observed for CYP2D1 by 32% (p < 0.01) after 10 days of treatment. No statistically significant differences were noted for CYPIA2, CYP2C6 and CYP3A2. In case of CYP3A1, the mRNA level of this gene was reduced by almost 35% (p < 0.05) both, after 3 and 10 days. CYP2D2 expression was also inhibited by the extract, but to a lesser degree when compared to CYP3A1. Moreover insignificant decrease of CYP2E1 expression level by 25% (p < 0.01) was observed after 3 days of treatment. CONCLUSIONS: These findings suggest that soybean extract may change the expression of CYP enzymes involved in biotransformation of xenobiotics (drugs, procarcinogens).

[Influence of standardized extract of Epilobium angustifolium on estrogen receptor alpha and beta expression in in vivo model]. / Wplyw standaryzowanego wyciqgu z Epilobium angustifolium na ekspresj mRNA receptorów estrogenowych alpha i beta w modelu in vivo.

OBJECTIVE: Evaluation of the influence of the standardized extract from the herb of Epilobium angustifolium on ERalpha and ERbeta mRNA expression in rat ventral prostate tissue and free serum estradiol level. MATERIALS AND METHODS: Rats were divided into 6 groups with 10 animals. ERalpha and ERbeta mRNA expression in rat ventral prostate tissue level was performed using real-time PCR method in Light Cycler system. Serum-free estradiol was evaluated using immunoenzymatic technique. RESULTS: In our experimental model there was an increase of ERa mRNA level by 9% and decrease by 36% of ERbeta mRNA level in ventral prostate tissue in rats administrated with testosterone and E. angustifolium extract, in comparison with testosterone alone administrated animals. CONCLUSIONS: E. angustifolium standardized extract influenced the expression of estrogen receptor alpha and beta mRNAs in differential manner which may suggest its potentially therapeutic properties or causing of adverse effects in pharmacotherapy of estrogen-related disorders. More complex studies should be undertaken to evaluate safety and to improve the efficacy of using this herbal extract.

[The influence of a standardized soybean extract (Glycine max) on the expression level of CYP3A enzymes and pregnane X receptor in in vivo model]. / Wplyw standaryzowanego ekstraktu sojoweg (Glycine max) na poziom ekspresji enzymów CYP3A i receptora pregnanu X w modelu in vivo.

OBJECTIVE: Soybean phytoestrogens, such as genistein and daidzein, have become a popular alternative for women undergoing the treatment of menopause symptoms. These isoflavones are also commonly used in traditional medicine in the prevention and treatment of osteoporosis, cardiovascular diseases and cancer Despite the widespread use of soybean preparations as functional foods and dietary supplements, data regarding the safety as well as interactions between herbal medicines and synthetic drugs, especially with antineoplastic agents, remain scarce. Therefore, the aim of the present study was to determine the influence of soybean extract on the expression levels of CYP3A and PXR genes using real-time PCR (RT-PCR). MATERIALS AND METHODS: Male Wistar rats were given a standardized soybean extract (100 mg/kg p.o.) for 3 and 10 days. Total RNA isolated from the liver tissue was transcribed into cDNA. The level of CYP3A 1/2 and PXR mRNAs expression was analyzed by real-time quantitative PCR using SYBR Green I dye. RESULTS: Our findings showed that soybean extract containing 37% isoflavones resulted in a significant decrease of CYP3A1 expression level by almost 35% (p<0.05), both after 3 and 10 days, when compared with the control group. No statistically significant differences were noted for CYP3A2 enzyme and the PXR nuclear factor. CONCLUSIONS: These results suggest that soybean extract can decrease the CYP3A1 (homolog to human CYP3A4) expression and may participate in clinically significant interactions with drugs metabolized by CYP3A4 enzyme. Moreover it is postulated that gene expression of CYP3A1 and CYP3A2 (homolog to human CYP3A5) can be regulated indirectly by the PXR transcription factor.