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  • Author: Bogusław Czerny x
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Summary

Introduction: Blood brain barrier and multidrug resistance phenomenon are subjects of many investigations. Mainly, because of their functions in protecting the central nervous system (CNS) by blocking the delivery of toxic substances to the brain. This special function has some disadvantages, like drug delivery to the brain in neurodegenerative diseases

Objective: The aim of this study was to examine how natural and synthetic substances affect the expression levels of genes (Mdr1a, Mdr1b, Mrp1, Mrp2, Oatp1a4, Oatp1a5 and Oatp1c1) that encode transporters in the blood-brain barrier.

Methods: cDNA was synthesized from total RNA isolated from rat hippocampus. The expression level of genes was determined using real-time PCR (RT-PCR) method.

Results: Our findings showed that verapamil, as a synthetic substance, caused the greatest reduction of mRNA level of genes studied. The standardized extract of Curcuma longa reduced the expression level for Mrp1 and Mrp2, whereas the increase of mRNA level was observed for Mdr1b, Oatp1a5 and Oatp1c1.

Conclusions: These results suggests that herbal extracts may play an important role in overcoming the blood brain barrier during pharmacotherapy.

Abstract

The aim of this study was to investigate the influence of standardized Epilobium angustifolium L. extract [100 mg/kg/day, p.o.] on the expression level of 5α-reductase type 2 (Srd5ar2) mRNA and Mapk3 mRNA a representative of non-genomic xenobiotics signaling pathway. It was shown that plant extract from the E. angustifolium showed a slight tendency to reduce prostate weight in hormonally induced animals (p>0.05) and in testosterone induced animals receiving both, extract and finasteride (p<0.05). Finasteride in rats induced by testosterone caused a smaller decrease in the level of mRNA 5α-steroid reductase 2 (SRd5ar2), than in rats treated with the hormone and studied plant extracts. In general, an increase in the amount of MAPK3 mRNAs in testosterone-induced groups of rats receiving tested plant extract with or without finasteride was observed, while the expression of type 2 5α-steroid reductase decreased (p<0.05). Further experimental studies should be performed in order to understand the molecular basis of interactions, the efficacy and safety of tested plant extracts.

Abstract

The aim of this study was to investigate the influence of standardized crude aqueous Epilobium angustifolium L. extract [100 mg/kg/day, p.o.] on the expression level of SRC kinase mRNA - a representatives of non-genomics xenobiotics signaling pathway in prostate ventral lobes of testosterone-induced, castrated rats. We have shown that in all analyzed groups induced by testosterone an elevation of SRC kinase mRNA transcription was observed, in comparison to control animals (not receiving the testosterone), (p<0.05). Finasteride in rats induced by testosterone caused the strongest inhibition of SRC mRNA transcription (p<0.05). In rats receiving testosterone and the plant extract a ca. 90% decrease of mRNA level was observed vs. testosterone-induced animals (p<0.05), while in testosterone-induced animals receiving concomitantly E. angustifolium extract and finasteride the observed reduction reached 87.3% (p<0.05).

We did not observed, however, any positive feedback between studied plant extract and finasteride in the inhibitory activity (p<0.05). Further experimental studies should be performed in order to the understanding the molecular basis of interactions, the efficacy and safety of tested plant extract.

Summary

In our research, the concentration of lotaustralin in the roots of two species Rhodiola kirilowii and Rhodiola rosea were compared. Aqueous and hydroalcoholic extracts from those plants were analyzed too. To determine the content of this compound the ultra performance liquid chromatography - tandem mass spectrometry (UPLC-MS/MS, Waters) was used. The obtained results showed that the content of measured lotaustralin depends on the species of Rhodiola. R. rosea roots are the richer source of lotaustralin then R. kirilowii. The same situation was observed in the extracts. A hydroalcoholic extract from R. rosea contains up to 135.276 mg of lotaustralin in 100 g of dry powdered material. In the case of R. kirilowii extracts, an aqueous extract contained more lotaustralin (74.791 mg/100 g of dry powdered material) then a hydroalcoholic extract.

Summary

The aim of the study was the identification and quantitative analysis of phenylpropanoid compounds in the roots of Rhodiola species. Rosavin, rosarin and rosin were determined in the roots of R. kirilowii and R. rosea from the field cultivation, Institute of Natural Fibres and Medicinal Plants. For the quantitative analysis, the ultra performance liquid chromatography - tandem mass spectrometry (UPLC-ESI MS/MS, Waters) was used. The results showed differences in the quantitative and qualitative assessments of these two species. In the root of R. kirilowii the presence of phenylpropanoids was not confirmed. In R. rosea the most common phenylpropanoid was rosavin (0.022%). The UPLC-MS/MS studies allowed to use this analytical method for determination of phenylpropanoids in the accordance with the requirements of ICH.

Summary

The aim of our study were qualitative and quantitative analyses of two polyphenolic acids: chlorogenic and gallic acids. These compounds were determined in two species of Rhodiola: R. kirilowii and R. rosea. After collecting plants, aqueous and hydroalcoholic extracts were prepared. In order to identify analysed polyphenolic compounds ultra performance liquid chromatography - tandem mass spectrometry (UPLC-MS/MS, Waters) was used. Gallic acid is commonly found in the roots of these plants. Aqueous extract in both species is a rich source of gallic acid. The UPLC-MS/MS studies allow to use this analytical method for determination of polyphenolic acids accordance with the requirements of ICH. Chromatographic method developed by our team is more precise then previously published.

Summary

Introduction: Despite widespread use of Panax ginseng and Ginkgo biloba, the data on the safety as well as herb-drug interactions are very limited. Therefore, we postulate that P. ginseng and G. biloba may modulate the activity and content of cytochrome P450 isozymes involved in the biotransformation of diverse xenobiotic substances. Objective: The aim of our study was to determine the influence of herbal remedies on the expression level of CYP enzymes and transcriptional factors. Methods: Male Wistar rats were given standardized Panax ginseng (30 mg/kg p.o.) or standardized Ginkgo biloba (200 mg/kg p.o.) for 3 and 10 days. The expression in liver tissue was analyzed by realtime PCR method. Results: Our results showed a decrease of CYP3A1 (homologue to human CYP3A4) mRNA level after P. ginseng extract treatment. The CYP2C6 (homologue to human CYP2C9) expression was also reduced. Additionally, after 10 days of the treatment with P. ginseng an increase of CYP1A1 (homologue to human CYP1A1) and CYP1A2 (homologue to human CYP1A2) expression was observed. Moreover, G. biloba extract also caused an increase of expression level for CYP1A1, CYP2C6, CYP3A1 and CYP3A2. Conclusion: Our findings suggest that herbal extracts can modulate the expression of transcriptional factors and CYP enzymes involved in xenobiotic metabolism and chemical carcinogenesis.

Summary

Introduction: Breast cancer is the most common malignant cancer among women. Both drug resistance and metastasis are major problems in the treatment of breast cancer. Therefore, adjuvant therapy may improve patients’ survival and affect their quality of life. It is suggested that epigallocatechin gallate (EGCG) which is well known for its chemopreventive activity and acts on numerous molecular targets may inhibit the growth and metastasis of some cancers. Hence, discovering the metastatic molecular mechanisms for breast cancer may be useful for therapy.

Objective: The aim of the study was to determine the effect of EGGC on the mRNA expression level of genes such as ZEB1, ABCB1, MDM2, TWIST1 and PTEN in MCF-7 breast cancer cells.

Methods: MCF7/DOX were cultured in the presence of 0.2 μM DOX and EGCG (20-50 μM). The mRNA expression level was determined by real-time quantitative PCR using RealTime ready Custom Panel 96 kit.

Results: Our results showed an important increase (about 2-fold for 20 μM EGCG + 0.2 μM DOX and 2.5-fold for 50 μM EGCG + 0.2 μM DOX, p<0.05) in ZEB1 expression levels. In case of ABCB1 gene lack of influence on the mRNA level was observed (p>0.05). We also observed significant decrease of ZEB1 expression in MCF7 cells with 20 μM and 50 μM EGCG (p<0.05). In addition, EGCG (20 μM) caused an increase of MDM2 and PTEN mRNA levels in almost 100% (p<0.05) and 40% (p>0.05), respectively. Lack of the influence of EGCG was noted for the TWIST1 gene expression. In case of MCF7/DOX we showed an increase of mRNA level of PTEN gene about 50% (p<0.05).

Conclusions: These results suggest that EGCG may be potentially used in adjuvant therapy in the breast cancer treatment.

Abstract

P-glycoprotein (P-gp) encoded by the MDR1 (multidrug resistance 1) gene is ATP-dependent transporting protein which is localizated in the cell membrane. P-gp is expressed mainly in organs with the secretory functions and its physiological role concerns tissue protection against xenobiotics. P-glycoprotein is involved in the permeability barriers of the blood-brain, blood-placenta directly protecting these organs. It participates in the transport of many drugs and other xenobiotics affecting their absorption, distribution, metabolism and excretion. The high P-gp activity in the cell membranes of cancer tissue is a major cause of lack of effectiveness of chemotherapy. Hence, the methods which could increase the sensibility of these pathological cells to cytostatics are still being searched. In the experimental studies it was shown that natural plant substances may have an effect on the expression level and activity of P-glycoprotein. Hypericum perforatum, Ginkgo biloba and Camellia sinensis increase P-gp activity while curcumin from Curcuma longa, piperine and silymarin inhibit this protein. Taking into account a wide substrate spectrum of P-gp, application of our knowledge on interactions of herbals and synthetic drugs should be considered in order to improve drug impact on different tissues.

Summary

Introduction: Plants are a rich source of healing substances. Cancer is a leading cause of death worldwide while breast cancer is the most common cancer among women. Circulating tumour cells (CTCs) are potential founder cells for metastasis. Therefore, their assessment may be used for monitoring of treatment as well as detecting cancer metastatis. Hence, it is suggested that the number of CTCs may be a valuable tumour biomarker during therapy.

Objective: The purpose of this study was to detect CTCs in breast cancer and to validate the method of assessment of CTC count using CytoTrack CT11 technology.

Methods: MCF-7 cells were sorted by a FACSARIA flow cytometer from blood samples derived from patients who have not been diagnosed with cancer. Identification and quantitative assessment of MCF-7 cells in blood samples were determined by flow sorting. Then, blood samples containing MCF-7 cells or without MCF-7 were scanned with the use of an automated fluorescence scanning microscope.

Results: In in vitro model analysing the glass CytoDisc™ with stained MCF-7 cells, we noted the correlation between the amount of observed tumour cells and expected number of tumour cells. Moreover, coefficient of variation in case of the recovery rate of the assumed number of MCF-7 cells was 30%, 17%, 18% and 15%, respectively.

Conclusion: Our study suggest that CTCs could be predictive factor in patients with metastatic cancer especially in breast cancer.