The expression of mRNAs for opioid precursors in endometrium of cyclic and early pregnant pigs; effects of IL-1β, IL-6 and TNFα

Expression of 3β-hydroxysteroid dehydrogenase and P450 aromatase in porcine oviduct during the oestrous cycle

“…The conditions of Real-Time PCR analysis were similar to these described by Wojciechowicz et al (2013) and Dziekoński et al (2015). The mean Ct values for each housekeeping gene were transformed using the comparative ΔCt method with the following equation: Q = E -ΔCt .…”

The expression of mRNAs for opioid receptors in endometrium of cyclic and early pregnant pigs; <i>in vitro</i> effects of IL-1β, IL-6 and TNFα

“…The animals were slaughtered on appropriate day of the cycle or pregnancy in a local abbatoir. The stage of the oestrous cycle was verified by testing morphology of the ovaries (Dziekoński et al, 2015), while pregnancy was confirmed by the presence of embryos in the uterine flushings. Uterus was placed in ice-cold phosphate buffered saline (PBS) supplemented with 100 IU·ml -1 of penicillin and 100 µg·ml -1 of streptomycin and transported to the laboratory for endometrial tissue isolation.…”

“…The quantity and purity of RNA were determined spectrophotometrically (TECAN; Männedorf, Switzerland), then randomly selected samples were additionally tested by the electrophoresis in 1.5% agarose gel. Reverse transcription (RT) was performed using QuantiTect Reverse Transcription Kit (Qiagen; Hilden, Germany) in the way described before (Dziekoński et al, 2015). Obtained cDNAs were frozen at -20°C and stored for Real-Time PCR analysis.…”

“…The presence of EOPs and/or the expression of opioid precursor genes in the uterine tissues has been demonstrated in many studies performed with females of different species, including pigs, as discussed in previous paper (Dziekoński et al, 2015). Information concerning the presence of opioid receptors in the uterus is scarce.…”

“…The expression of prodynorphin (PDYN) gene was inhibited by and TNFα in the human macrophage U-937 line (Sun et al, 2006), however, IL-1 was not effective in this study. In our recent study, the effects of selected cytokines (IL-1β, IL-6 and TNFα) on the expression of genes encoding opioid precursors in the porcine endometrial explants on days 10-11, 12-13, 15-16 of the oestrous cycle and early pregnancy were observed (Dziekoński et al, 2015). The strongest influence was exerted by IL-6 on the PENK gene expression during all studied days of the cycle as well as on days 10-11 and 12-13 of pregnancy.…”