Problem
Cytokines, mediators of the immune response, are involved in the regulation of female reproductive processes during the estrous cycle and pregnancy. The present study aimed to investigate the effect of selected peroxisome proliferator‐activated receptor gamma (PPARγ) ligands on the expression of nuclear factor kappa B (NF‐κB) and selected cytokines, such as interleukin (IL)‐1β, ‐4, ‐6, ‐8, ‐10, and the leukemia inhibitory factor, in the porcine endometrium on days 10‐12 and 14‐16 of the estrous cycle (mid‐ and late luteal phase, respectively) or pregnancy (maternal recognition of pregnancy and beginning of implantation, respectively).
Method of study
Endometrial slices were incubated in vitro in the presence of PPARγ agonists, 15‐deoxy‐Δ12, 14‐prostaglandin J2 or rosiglitazone, and PPARγ antagonist T0070907. mRNA and protein levels in tissues were determined by real‐time PCR and Western blot.
Results
On days 10‐12 of the estrous cycle and days 14‐16 of pregnancy, PPARγ ligands enhanced the expression of NF‐κB, mRNA cytokines, and/or proteins. During the late luteal phase of the estrous cycle (days 14‐16) and maternal recognition of pregnancy (days 10‐12), PPARγ ligands inhibited the expression of NF‐κB, and they differentially affected the expression of mRNA and proteins of cytokines.
Conclusion
Our results indicate that PPARγ is engaged in the endometrial synthesis of NF‐κB and selected cytokines in pigs. The influence of PPARγ ligands on the tested components of the immune system varied subject to the physiological status of females, and it could be associated with differences in endometrial receptivity.
Female fertility depends greatly on the capacity of the uterus to recognize and eliminate microbial infections, a major reason of inflammation in the endometrium in many species. This study aimed to determine the in vitro effect of PPARγ ligands on the transcriptome genes expression and alternative splicing in the porcine endometrium in the mid-luteal phase during LPS-stimulated inflammation using RNA-seq technology. The endometrial slices were incubated in vitro in the presence of LPS and PPARγ agonists—PGJ2 or pioglitazone and antagonist—T0070907. We identified 222, 3, 4 and 62 differentially expressed genes after LPS, PGJ2, pioglitazone or T0070907 treatment, respectively. In addition, we detected differentially alternative spliced events: after treatment with LPS—78, PGJ2–60, pioglitazone—52 or T0070907–134. These results should become a basis for further studies explaining the mechanism of PPARγ action in the reproductive system in pigs.
Inflammation is a biological response of the immune system, which can be triggered by many factors, including pathogens. These factors may induce acute or chronic inflammation in various organs, including the reproductive system, leading to tissue damage or disease. In this study, the RNA-Seq technique was used to determine the in vitro effects of peroxisome proliferator-activated receptor gamma (PPARγ) ligands on the expression of genes and long non-coding RNA, and alternative splicing events (ASEs) in LPS-induced inflammation of the porcine endometrium during the follicular phase of the estrous cycle. Endometrial slices were incubated in the presence of LPS and PPARγ agonists (PGJ2 or pioglitazone) and a PPARγ antagonist (T0070907). We identified 169, 200, 599 and 557 differentially expressed genes after LPS, PGJ2, pioglitazone or T0070907 treatment, respectively. Moreover, changes in differentially expressed long non-coding RNA and differential alternative splicing events were described after the treatments. The study revealed that PPARγ ligands influence the LPS-triggered expression of genes controlling the DNA damage response (GADD45β, CDK1, CCNA1, CCNG1, ATM). Pioglitazone treatment exerted a considerable effect on the expression of genes regulating the DNA damage response.
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