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The expression of mu (μ), delta (δ) and kappa (κ) opioid receptors was determined in vitro under the basal conditions as well as in the presence of cytokines (interleukin (IL)-1β, IL-6 and tumor necrosis factor α (TNFα)) in the endometrium of gilts on days 10–11, 12–13 and 15–16 of the oestrous cycle and pregnancy. These days were chosen due to their importance for the establishment of pregnancy in pigs. During the oestrous cycle, the basal expression of μ opioid receptor mRNA in endometrial explants was greater on days 10–11 than on days 12–13 and 15–16, during pregnancy its expression was greater on days 12–13 than on days 10–11 and 15–16. The expression of δ opioid receptor mRNA did not change during the cycle, but during pregnancy it was the greatest on days 10–11. The expression of κ opioid receptors did not significantly change during studied periods. The cytokines affected the expression of μ and δ, but not κ opioid receptors in endometrial explants. The expression of μ opioid receptor mRNA was significantly decreased by IL-1β on days 10–11, but IL-6 increased and decreased it on days 12–13 and 15–16 of the cycle, respectively. During pregnancy, the expression of μ opioid receptor mRNA in endometrial explants was increased by IL-1β, IL-6 and TNFα on days 10–11, decreased by IL-1β on days 12–13, and increased by TNFα on days 15–16. During the oestrous cycle, the expression of δ opioid receptor mRNA was elevated by IL-1β on all studied days, by IL-6 on days 10–11 and 12–13, and by TNFα on days 15–16. During pregnancy, IL-1β increased the expression of δ opioid receptor transcript on days 12–13. These data suggest that opioid receptors participate in a local regulation of uterine functions in gilts during the oestrous cycle and early pregnancy and their endometrial expression can be modulated by cytokines.
The expression of genes encoding opioid precursors: proopiomelanocortin (POMC), proenkephalin (PENK) and prodynorphin (PDYN) was studied in the porcine endometrial explants in the presence or absence of selected cytokines (interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor α (TNFα)) on days 10–11, 12–13, 15–16 of the oestrous cycle and pregnancy. In pregnant pigs, these days among others are important for 1. migration of the embryos, 2. maternal recognition of pregnancy and 3. the onset of implantation, respectively. During the oestrous cycle, the basal (in the absence of cytokines) mRNA expression of PDYN, but not POMC or PENK was increased on days 15–16 compared to days 10–11. During pregnancy, the gene expression of POMC increased on days 10–11 and 15–16 (vs days 12–13), but PENK – on days 12–13 and 15-16 (vs days 10–11) as well as PDYN – on days 15-16 (vs days 12–13) decreased. The endometrial expression of studied genes was influenced by cytokines. The expression of PENK was stimulated by IL-6 in all studied days of the cycle as well as on days 10–11 and 12–13 of pregnancy. In turn, the PENK mRNA expression was increased by TNFα on days 10–11 and decreased by IL-1β on days 15–16 of pregnancy. The POMC expression was decreased by IL-1β on days 12–13, but increased by both IL-6 and TNFα on days 15–16 of the cycle. The PDYN mRNA expression in endometrial explants was increased by IL-1β on days 10–11 of the cycle. These results indicate that the opioid system is involved in the local regulation of uterine functions during the oestrous cycle end early pregnancy in pigs.
In gilts, the period of early pregnancy occurring from the time of fertilization to the beginning of implantation is sensitive to any environmental disruptions, including an unbalanced diet of a future mother. Previously, we found that due to the undernutrition in gilts during this period, the endocrine intrauterine microenvironment and DNA methylation in the uterus have been changed. These distortions may diminish the success of pregnancy. In this study we focused on the influence of a restricted diet used in gilts during the first days of pregnancy on their biochemical and haematological parameters in peripheral blood. The applied restrictive diet vs. normal diet covered only 70% of the nutritional demands of early pregnant gilts. Normal (n = 4 gilts) or restrictive (n = 5 gilts) diets were used from the day of the first signs of the estrus until day 9 of pregnancy and biochemical and haematological parameters in blood plasma were determined during peri-implantation period, e.g. on days 15 to16 of pregnancy. In restrictive vs. normal fed gilts significantly lower plasma phosphorus, calcium and total cholesterol as well as the tendency to increasing concentrations of triglicerydes and asparate aminotranserase were found. Haematological parameters did not differ between the studied gilts. Thus, it seems that the availability of nutritional factors became suboptimal in restrictively fed early pregnant gilts. Even short-lasting undernutrition of females during the peri-conceptional period may cause a disruption of biochemical homeostasis during the peri-implantation period and probably affect the success of pregnancy.
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