Propolis is a natural product that honeybees collect from various plants. It is known for its beneficial pharmacological effects. The aim of our study was to evaluate the impact of propolis on human sperm motility, mitochondrial respiratory activity, and membrane potential. Semen samples from 10 normozoospermic donors were processed according to the World Health Organization criteria. Propolis effects on the sperm motility and mitochondrial activity parameters were tested in the fresh ejaculate and purified spermatozoa. Propolis preserved progressive motility of spermatozoa in the native semen samples. Oxygen consumption determined in purified permeabilized spermatozoa by high-resolution respirometry in the presence of adenosine diphosphate and substrates of complex I and complex II (state OXPHOSI+II) was significantly increased in the propolis-treated samples. Propolis also increased uncoupled respiration in the presence of rotenone (state ETSII) and complex IV activity, but it did not influence state LEAK induced by oligomycin. Mitochondrial membrane potential was not affected by propolis. This study demonstrates that propolis maintains sperm motility in the native ejaculates and increases activities of mitochondrial respiratory complexes II and IV without affecting mitochondrial membrane potential. The data suggest that propolis improves the total mitochondrial respiratory efficiency in the human spermatozoa in vitro thereby having potential to improve sperm motility.
- Publication type
- Journal Article MeSH
x
- MeSH
- Immunohistochemistry MeSH
- Embryonal Carcinoma Stem Cells * MeSH
- Cerebellum MeSH
- Mice, Neurologic Mutants MeSH
- Mice, Inbred CBA MeSH
- Tumor Cells, Cultured MeSH
- Neural Stem Cells * MeSH
- Graft Survival * MeSH
- Purkinje Cells cytology MeSH
- Stem Cell Transplantation * MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
- Comparative Study MeSH
Cíl studie: Jednou z příčin mužské neplodnosti je snížená motilita spermií. Ukazuje se, že ve vývoji této poruchy může hrát roli snížená efektivita respirační aktivity mitochondrií. Cílem naší studie bylo komplexní stanovení respirační aktivity mitochondrií spermií s normální a sníženou pohyblivostí. Typ studie: Prospektivní studie. Název a sídlo pracoviště: Ústav histologie a embryologie, LF UK, Plzeň; Ústav fyziologie, LF UK, Plzeň; Institut reprodukční medicíny a endokrinologie, IVF Centrum Prof. Zecha, Plzeň. Metodika: Ejakuláty byly získány od 14 mužů z IVF Centra Prof. Zecha v Plzni. Podle klasifikace World Health Organization byli muži rozděleni do skupiny normozoospermatiků (n = 7) a astenozoospermatiků (n = 7). Respirační aktivitu spermií jsme měřili na dvoukanálovém oxygrafu Oroboros. Výsledky: V astenozoospermatických vzorcích byla nalezena signifikantně snížená aktivita komplexu I(p = 0,007), zvýšená respirace po aplikaci inhibitoru ATP-syntázy oligomycinu (ukazující na zvýšené rozpřažení oxidace a fosforylace; p = 0,046). Inhibice komplexu I rotenonem ukázala, že příspěvek komplexu I k celkové kapacitě oxidační fosforylace byl i u zdravých spermií relativně nižší, než je tomu typicky v somatických buňkách. Závěr: V naší studii jsme měřili respirační aktivitu mitochondrií lidských spermií permeabilizovaných digitoninem vysokoúčinnou oxygrafií, která umožňuje stanovení spotřeby kyslíku z nejmenšího možného množství zárodečných buněk. Výsledky studie potvrzují sníženou aktivitu komplexu I u astenozoospermatiků a naznačují, že na snížené pohyblivosti spermií by se mohl podílet i zvýšený únik protonů z mitochondriální matrix, který vede ke snížené efektivitě fosforylačního procesu. Lepší charakterizace mužských zárodečných buněk, ať zcela zdravých, či s postiženou motilitou, nám pomůže lépe pochopit proces fyziologického oplodnění a zároveň pomůže i ve výběru té nejvíce životaschopné spermie pro léčbu neplodnosti metodami asistované reprodukce.
Objective: One of causes of male infertility is reduced sperm motility. It turns out that the reduced efficiency of the mitochondrial respiratory activity may play a role in the development of this disorder. The aim of our study was to comprehensively determine mitochondrial respiratory activity of sperm with normal and reduced motility. Design: Prospective study. Setting: Department of Histology and Embryology, Faculty of Medicine in Pilsen, Charles University in Prague; Department of Physiology, Faculty of Medicine in Pilsen, Charles University in Prague; Institute of Reproductive Medicine and Endocrinology, IVF Centers Prof. Zech, Plzeň. Methods: Ejaculates of 14 men were obtained from IVF Center Prof. Zech, Pilsen. According to the World Health Organization classification, samples were divided into normozoospermatic (n = 7) and asthenozoospermatic(n = 7) groups. Respiratory activity of sperm was measured on two-chamber oxygraph Oroboros. Results: In asthenozoospermatic samples, significantly reduced activity of complex I (p = 0.007) and increased respiration after application of ATP-synthase inhibitor oligomycin (showing increased uncoupled oxidation and phosphorylation, p = 0.046) were found. Inhibition of complex I by rotenone showed that complex I contribution to the total capacity of oxidative phosphorylation of healthy sperm was relatively lower than it is typical for somatic cells. Conclusion: In our study, we measured mitochondrial respiratory activity of human sperm, permeabilized by digitonin, by high-resolution oxygraphy, which allows the determination of oxygen consumption from the smallest possible number of germ cells. The study results confirm reduced activity of complex I in asthenozoospermatics and suggest that increased leakage of protons from the mitochondrial matrix, which leads to reduced efficiency of phosphorylating process, could participate in the reduced sperm motility. Better characterization of male germ cells, either completely healthy or with affected motility, will help us to understand better the physiological process of fertilization and also to choose the most viable sperm for infertility treatment by methods of assisted reproduction.
- MeSH
- Asthenozoospermia metabolism MeSH
- Clinical Laboratory Techniques MeSH
- Humans MeSH
- Mitochondrial Proteins * physiology metabolism MeSH
- Sperm Motility physiology MeSH
- Infertility, Male MeSH
- Cell Membrane Permeability MeSH
- Policy MeSH
- Reactive Oxygen Species * metabolism MeSH
- Semen * metabolism MeSH
- Spermatozoa * physiology pathology MeSH
- Oxygen Consumption MeSH
- Statistics as Topic MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
Cíl studie: Obezita i nadváha negativně ovlivňují schopnost ženy přirozeně otěhotnět, podílí se na zvýšeném výskytu porodnických komplikací v průběhu těhotenství a negativně ovlivňují i výsledek technik asistované reprodukce (ART). Cílem naší práce bylo porovnat výsledky léčby neplodnosti u skupiny neplodných žen podstupujících ART v závislosti na hodnotách jejich BMI a změny hladin vybraných hormonů a markerů oxidačního stresu ve folikulární tekutině. Typ studie: Retrospektivní komparativní studie. Název a sídlo pracoviště: LF UK, Plzeň; Institut reprodukční medicíny a endokrinologie, IVF Centrum Prof. Zecha, Plzeň. Metodika: Studovaný soubor tvořilo 44 žen (průměrný věk 31,9 let, SD = 4,35) léčených pro neplodnostv Institutu reprodukční medicíny a endokrinologie – IVF Centrum Prof. Zecha. Ženy byly rozděleny do dvou skupin podle hodnoty BMI (37 žen mělo normální BMI, 7 žen mělo nadváhu). Hladiny prolaktinu, volného T3 a T4 hormonu, homocysteinu, malondialdehydu, glutathionperoxidázy (GPx), celkové antioxidační kapacity a celkové bílkoviny byly analyzovány ve folikulární tekutině u obou sledovaných skupin. U každé ženy byly použity vzorky folikulární tekutiny bez příměsi krvea ze všech foliklů dohromady, tedy ze společného poolu. Výsledky: Pozorována byla jen signifikantně nižší hladina GPx u skupiny žen s nadváhou (p = 0,0044). V úspěšnosti pregnancy rate se ženy s normálním BMI a ženy trpící nadváhou od sebe významně nelišily(p = 0,4430). Závěr: Naše studie nepotvrdila negativní efekt nadváhy na výsledky léčby neplodnosti, konkrétně pregnancy rate u skupiny neplodných žen podstupujících léčbu pomocí technik asistované reprodukce.
Objective: Obesity and overweight negatively affect the ability of women to conceive naturally, contributes to the increased incidence of obstetric complications during pregnancy and affect the outcome of assisted reproduction techniques (ART). The aim of our study was to compare the results of treatment of infertilityin a group of infertile women undergoing ART, depending on the values of BMI and changes in levels of selected hormones and markers of oxidative stress in follicular fluid. Design: Retrospective comparative study. Setting: Faculty of Medicine in Pilsen, Charles University in Prague; Institute of Reproductive Medicine and Endocrinology, IVF Centers – Prof. Zech, Plzeň. Methods: The studied group consisted of 44 women (mean age of 31.9 years, SD = 4.35) treated for infertility at the Institute of Reproductive Medicine and Endocrinology – IVF Centers Prof. Zech. Women were divided into 2 groups according to BMI (37 women had normal BMI, 7 women were overweight). Prolactin, free T3 and T4 hormone, homocysteine, malondialdehyde, glutathione peroxidase, total antioxidant capacity and total protein were analyzed in the follicular fluid both groups. Only blood free samples were studied after pooling of all FF samples of each patient. Results: We observed significantly lower levels of glutathion peroxidase in the group of overweight women(p = 0.0044). The pregnancy success rate with women with normal BMI and overweight women did not differ significantly from each other (p = 0.4430). Conclusion: Our study did not confirm the negative effect of obesity on the results of treatment of infertility, specifical-ly pregnancy rate in a group of infertile women undergoing treatment with assisted reproduction techniques.
- MeSH
- Reproductive Techniques, Assisted * MeSH
- Adult MeSH
- Follicular Fluid * physiology metabolism MeSH
- Glutathione Peroxidase diagnostic use MeSH
- Thyroid Hormones MeSH
- Body Mass Index MeSH
- Humans MeSH
- Malondialdehyde MeSH
- Overweight * metabolism MeSH
- Obesity metabolism MeSH
- Oxidative Stress MeSH
- Prolactin MeSH
- Retrospective Studies MeSH
- Statistics as Topic MeSH
- Infertility, Female * etiology physiopathology therapy MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Female MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
Cíl studie: Folikulární tekutina (FT) zajišťuje charakteristické a nezastupitelné mikroprostředí, které ovlivňuje vývoj oocytu. Biochemické složení FT má zásadní vliv na vývojový potenciál vajíčka. Cílem naší práce bylo srovnání hladin vybraných hormonů v FT žen s poruchou plodnosti a zdravých plodných dárkyň oocytů. Typ studie: Retrospektivní komparativní studie. Název a sídlo pracoviště: Ústav histologie a embryologie, LF UK a FN v Plzni; Gynekologicko-porodnická klinika FN a LF UK v Plzni; Institut reprodukční medicíny a endokrinologie, IVF Centra Prof. Zech, Plzeň. Metodika: Hladiny prolaktinu, volného T3 (fT3) a volného T4 (fT4) hormonu byly analyzovány v FT u obou sledovaných skupin. Celkem jsme vyšetřili 146 žen – 74 pacientek léčených pro neplodnost (průměrný věk 31 let, SD = 4,65) a 72 zdravých plodných žen, dárkyň oocytů (průměrný věk 26 let, SD = 4,44). U každé ženy byly použity vzorky FT bez příměsi krve a ze všech folikulů dohromady, tedy ze společného poolu. Hladiny hormonů byly stanoveny pomocí metodiky ECLIA (elektrochemiluminiscenční imunoanalýza) na přístroji Cobas e411. Výsledky: Výsledky analýz prokázaly statisticky signifikantně vyšší hladiny prolaktinu (p = 0,0006) a volného T4 hormonu (p=0,0246) u žen s poruchou plodnosti oproti skupině zdravých plodných dárkyň oocytů. Závěr: Naše studie potvrzuje přítomnost prolaktinu a tyroideálních hormonů ve FT a naznačuje jejich klíčovou roli v regulaci reprodukčních procesů. Další studium dárkyň oocytů a jejich podrobnější srovnání s neplodnými pacientkami s různými gynekologickými příčinami poruch plodnosti bude přínosem pro objasnění patofyziologických mechanismů regulujících plodnost.
Objective: Follicular fluid (FF) provides an important microenvironment for the development of oocytes. The biochemical composition of the FF plays critical role in the oocyte competence. The aim of our study was to compare the levels of selected hormones in FF of infertile women and healthy fertile oocyte donors. Design: Retrospective comparative study. Setting: Department of Histology and Embryology, Faculty of Medicine in Pilsen, Charles University in Prague; Department of Gynecology and Obstetrics, Charles University and University Hospital in Pilsen; Institute of Reproductive Medicine and Endocrinology, IVF Centers Prof. Zech, Plzeň. Methods: Levels of prolactin, free T3 and free T4 hormones in the FF of 146 women were analyzed. We have analysed FF of 74 infertile patients (mean age 31 years, SD = 4.65) and 72 healthy fertile oocyte donors (mean age 26 years, SD = 4.44). Only blood free samples were studied after pooling of all FF samples each patient. Levels of hormones were determined using ECLIA method (Electro-Chemi-Luminiscent Immunoassay) on the Cobas e411. Results: Results showed statistically significantly higher levels of prolactin (p=0.0006) and free T4 hormone (p=0.0246) in FF of infertile women in comparison to the group of healthy fertile oocyte donors. Conclusion: Our study confirms the presence of prolactin and thyroid hormones in FF and it can be suggested that they play a key role in the regulation of reproductive processes. The study of FF from donors and their detailed comparison with infertile patients with various gynaecological causes of infertility has great value for better understanding of regulatory mechanisms of fertility.
- Keywords
- volný T4, volný T3,
- MeSH
- Oocyte Donation MeSH
- Adult MeSH
- Financing, Organized MeSH
- Follicular Fluid MeSH
- Thyroid Hormones * analysis blood MeSH
- Humans MeSH
- Statistics, Nonparametric MeSH
- Prolactin * analysis blood MeSH
- Retrospective Studies MeSH
- Triiodothyronine analysis blood MeSH
- Infertility, Female * MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Female MeSH
- Publication type
- Comparative Study MeSH
Cíl studie: Vývoj oocytu je ovlivňován prostředím, zvláště folikulární tekutinou (FT). Narušení její homeostázy má zásadní vliv na vývojový potenciál vajíčka. Cílem této studie bylo srovnání hladin vybraných markerů oxidačního stresu ve FT žen s poruchou plodnosti a zdravých plodných dárkyň oocytů. Typ studie: Retrospektivní komparativní studie. Název a sídlo pracoviště: Lékařská fakulta v Plzni, Univerzita Karlova v Praze; Institut reprodukční medicíny a endokrinologie, IVF Centra Prof. Zech, Plzeň. Metodika: Ve FT byly analyzovány hladiny homocysteinu (HCy), malondialdehydu (MDA), glutathionperoxidázy (GPx), celkové antioxidační kapacity (AOK) a celkové bílkoviny (CB). Celkem bylo vyšetřeno 146 žen – 74 pacientek léčených pro neplodnost (průměrný věk 31 let, SD = 4,65) a 72 zdravých plodných žen, dárkyň oocytů (průměrný věk 26 let, SD = 4,44). U každé ženy byly použity vzorky FT ze všech folikulů dohromady, tedy ve společném poolu. Ze studie byly vyloučeny vzorky, ve kterých se vyskytla příměs krve. Výsledky: Studie prokázala statisticky signifikantně vyšší hladiny HCy (p < 0,0001) ve FT u zdravýchplodných žen oproti skupině s poruchou plodnosti, a to jak bez ohledu na věk, tak ve skupinách neplodných kontrol ve stejném věkovém rozmezí (u věkové skupiny mezi 20 až 29 lety jep = 0,0002, ve věku 30 až 39 let je p < 0,0001). Při rozdělení souborů do výše popsaných věkových pásem jsme prokázali statisticky signifikantně vyšší hladinu MDA u kontrolní skupiny ve věku 20 až 29 let oproti stejně starým ženám léčeným pro neplodnost (p = 0,0374) a statisticky signifikantně vyšší hodnotu AOK u neplodných žen mezi 30. až 39. rokem života oproti zdravým plodným dárkyním oocytů(p = 0,0458). Závěr: Přítomnost, či naopak nepřítomnost prooxidačních parametrů ve FT má důležitou úlohuve schopnosti koncepce a následném vývoji embrya. Klíčová slova: folikulární tekutina, homocystein, oxidační stres, neplodnost, dárkyně oocytů.
Objective: Follicular fluid (FF) affects oocyte development and disruption of its homeostasis has a crucial effect on egg developmental potential. The aim of this study was to compare the levels of selected oxidative stress markers in the FF of women with impaired fertility and healthy fertile oocytes donors. Design: A retrospective comparative study. Setting: Faculty of Medicine in Pilsen, Charles University in Prague; Institute of Reproductive Medicine and Endocrinology, IVF Center Prof. Zech, Pilsen. Methods: Levels of homocysteine (Hcy), malondialdehyde (MDA), glutathione peroxidase (GPx), total antioxidant capacity (AOK) and total protein (CB) were analyzed in the FF. We have analysed FF of 146 women – 74 infertile patients (mean age 31 years, SD = 4.65) and 72 healthy fertile oocyte donors (mean age 26 years, SD = 4.44). Only blood free samples were studied after pooling of all FF samples each patient. Results: The study showed a statistically significantly higher Hcy levels (p < 0.0001) in the FF of healthy fertile women compared with impaired fertility group both – comparing the two groups regardless the age and in groups of the same age range (for the age group between 20 to 29 years isp = 0.0002, for the age group between 30 to 39 years is p < 0.0001). When divided into above age ranges we found statistically significantly higher levels of MDA in the control group aged 20 to 29 years compared to same age infertile patients (p = 0.0374) and statistically significantly higher AOK in infertile women between 30 to 39 years of age compared to same age control group (p = 0.0458). Conclusion: The presence or on the contrary the absence of prooxidant parameters in the FF has an important role in the ability of conception and subsequent embryo development. Key words: follicular fluid, homocysteine, oxidative stress, infertility, oocyte donor.
- MeSH
- Analysis of Variance MeSH
- Oocyte Donation * MeSH
- Adult MeSH
- Follicular Fluid * MeSH
- Glutathione Peroxidase analysis MeSH
- Homocysteine analysis MeSH
- Data Interpretation, Statistical MeSH
- Humans MeSH
- Malondialdehyde analysis MeSH
- Statistics, Nonparametric MeSH
- Oxidative Stress * physiology MeSH
- Retrospective Studies MeSH
- Infertility, Female * MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Female * MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
- Comparative Study MeSH
Mouse embryonic carcinoma cells (P19 line) were studied for both their survival and developmental potential in the intact cerebellum of B6CBA mice. The P19 cells were cultured and labelled with green fluorescent protein using transfection. Cells were used for transplantation either in the undifferentiated stage or after 3 days of neurodifferentiation induced by retinoic acid. The intracerebellar application was performed in 43 mice: group A (N = 21) received neuroprogenitors and group B (N = 22) received undifferentiated cells. The morphology of transplanted cells within the context of the surrounding cerebellar tissue was evaluated after 3 weeks. Naive P19 cells engrafted and survived in the cerebellum of 7 of the 22 adult mice (survival rate 31.8 %). Neuroprogenitors survived in 13 of the 21 mice (survival rate was 61.9 %). Since the cut-off is P < 0.05, the difference is not statistically significant (P = 0.069). An expansive appearance of the graft was significantly more frequent (P = 0.0047) in naive P19 cells than in neuroprogenitors. In mice in which the grafts did not survive, no marks of grafted cells or only fluorescing detritus were found. In conclusion, this is the first study to track the fate and morphology of embryonic carcinoma cells transplanted into the cerebellum, confirming that neuroprogenitors derived from embryonic carcinoma cells can settle in the host tissue and differentiate according to the surrounding conditions. With further validation, the embryonic carcinoma cells could become a valuable model with which to study the impact of cell therapy on neurodegenerative diseases.
- MeSH
- Cell Differentiation physiology MeSH
- Fluorescent Antibody Technique, Indirect MeSH
- Stem Cells cytology MeSH
- Cerebellum cytology MeSH
- Mice MeSH
- Cell Line, Tumor MeSH
- Neurons cytology MeSH
- Reverse Transcriptase Polymerase Chain Reaction MeSH
- Cell Transplantation MeSH
- Stem Cell Transplantation MeSH
- Cell Survival physiology MeSH
- Blotting, Western MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
- MeSH
- Cell Differentiation drug effects MeSH
- Cell Line cytology MeSH
- Embryonic Stem Cells cytology metabolism drug effects MeSH
- Immunohistochemistry methods MeSH
- Embryonal Carcinoma Stem Cells cytology metabolism drug effects MeSH
- MAP Kinase Kinase 2 analysis MeSH
- Mice MeSH
- Neurons cytology metabolism MeSH
- Pilot Projects MeSH
- In Vitro Techniques MeSH
- Tretinoin pharmacokinetics pharmacology MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
The aim of our study was to characterize mouse embryonal carcinoma (EC) cells P19 in different stages of retinoic acid induced neurodifferentiation by two methods, immunocytochemistry and RT qPCR. The characterization of the cells is crucial before any transplantation into any model, e.g. in our case into the mouse brain with the aim to treat a neurodegenerative disease. Specific protein markers (MAP-2, OCT-4, FORSE-1) were detected by immunocytochemistry in the cell cultures. The mRNA expression levels of PAX-6, MASH-1, Brachyury, GATA-4 and AFP were determined by RT qPCR method. HPRT was used as a housekeeping gene. The degree of differentiation can be characterized by expression of analyzed genes. The presence of OCT-4 and FORSE-1 proteins in undifferentiated pluripotent cells and the presence of dendrite specific MAP-2 in neuroprogenitors was detected. The expression levels of PAX-6 and MASH-1 increased and expression of Brachyury decreased during the neurodifferentiation process. The expression levels of GATA-4 and AFP were the highest after induction of differentiation with retinoic acid. Detailed characterization of cells before transplantation experiments can contribute to better understanding of their effect.
- MeSH
- Antigens, Surface genetics metabolism MeSH
- Cell Differentiation genetics drug effects MeSH
- Gene Expression MeSH
- Financing, Organized MeSH
- Homeodomain Proteins genetics metabolism MeSH
- Immunohistochemistry MeSH
- Embryonal Carcinoma Stem Cells physiology MeSH
- Mice MeSH
- Cell Line, Tumor MeSH
- Tumor Cells, Cultured MeSH
- Eye Proteins genetics metabolism MeSH
- Octamer Transcription Factor-3 genetics metabolism MeSH
- Reverse Transcriptase Polymerase Chain Reaction MeSH
- Microtubule-Associated Proteins genetics metabolism MeSH
- Repressor Proteins genetics metabolism MeSH
- Basic Helix-Loop-Helix Transcription Factors metabolism MeSH
- Paired Box Transcription Factors genetics metabolism MeSH
- Tretinoin pharmacology MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH