Short communication: Proinflammatory gene expression relative to the collection technique of endometrial samples from cows with and without subclinical endometritis

Abstract

Uterine inflammation negatively affects reproductive performance and is an important cause of infertility and subfertility in dairy cows. Several studies have investigated the use of gene expression in endometrial samples collected by biopsy or cytology to evaluate the inflammatory response of the cow uterus. This study aimed to compare the expression of the CCL5, CXCL8, IL6, and IL1B genes in the bovine endometrium according to the site of sample collection [caruncular (C) or intercaruncular (IC)], the collection method (biopsy or cytology), and the category of inflammation based on endometrial cytology (zero, medium, or high) in subclinical endometritis. The reproductive tracts of dairy cows were collected from a slaughterhouse, and punch-biopsy samples of endometrial tissues were obtained from both regions (C and IC). Endometrial cells from these regions were collected with the cytobrush technique and then used for the analysis of mRNA expression by quantitative PCR. After counting polymorphonuclear cells (PMN) by endometrial cytology, 20 uteri with an ovary at stage I (d 1-4 of estrous cycle) were categorized into 3 groups. Uteri with 0% PMN (n = 10) were assigned to group zero, uteri with 5 to 15% PMN (n = 5) to group medium (12.2 ± 1.6% PMN), and uteri with >15% PMN (n = 5) to group high (53.8 ± 32.9% PMN). All data were analyzed with 2-way ANOVA with Bonferroni multiple comparison post test. The results from gene transcripts demonstrated that the region (C or IC) of the endometrial biopsy had no influence on any of the degrees of inflammatory reaction observed. However, gene expression was more elevated in the endometrium of cows with greater inflammation compared with those without inflammation (CCL5, CXCL8, IL6, IL1B) and those with medium inflammation (CCL5, IL6). Expression of the genes evaluated did not differ between the endometrium without inflammation and with medium inflammation. However, in the high inflammation group, all genes were comparatively more expressed in samples collected by cytology relative to those derived from biopsies for both anatomical regions. In conclusion, gene expression did not differ between the C and IC tissue. Samples collected from animals with greater inflammation had greater gene expression than those with zero or medium inflammation. In addition, cytology samples had greater gene expression than biopsy samples in the high inflammation group.

Effect of Retained Placenta And Clinical Mastitis On Reproduction Parameters, Immune Response And Steroidogenic Receptors Gene Expression In Postpartum Crossbred Crossbreed Dairy Cows

“…In contrast,Shaheen et al (2020) observed an increase in the levels of cytokines as IL-2 and IL-6 in lactating crossbreed cows diagnosed with CM. Interestingly, no difference in the expression of chemokine CCL5 was observed between healthy, RP and CM groups Fagundes et al (2019). found no differences in the expressions of CCL5 in cows when caruncular and intercaruncular regions of the endometrial epithelium were compared in cases of absent or mild in ammation; however, the expression was higher in endometrium with severe in ammation when compared to groups with mild in ammation or none.In this study, we aimed to nd a relationship between the expression of steroidogenic genes andpostpartum immune function.…”

Subclinical endometritis differentially affects the transcriptomic profiles of endometrial glandular, luminal, and stromal cells of postpartum dairy cows

“…When using the same in vitro model as Guo et al (2019), changes in proteins related to the above pathways were also observed (Piras et al, 2017). In vivo, cows with SCE present increased local gene expression of proinflammatory cytokines characteristic of acute inflammatory processes (Kasimanickam et al, 2014;Fagundes et al, 2019;Pereira et al, 2020). In addition, cows affected by endometritis exhibit a large number of genes involved in immune response, cell adhesion, chemotaxis, apoptosis, and G-protein coupled receptor signaling pathways differentially expressed compared with healthy cows (Salilew-Wondim et al, 2016).…”

“…However, to date, gene expression studies aiming to analyze the effects of SCE from in vivo material were either based on biopsy-derived whole-endometrium tissue (Salilew-Wondim et al, 2016;Raliou et al, 2019) or endometrial swab or cytobrush samples (Kasimanickam et al, 2014;Fagundes et al, 2019), without discriminating between endometrial cell types. The bovine endometrium is a complex and heterogeneous tissue, and quantification of gene transcription from the whole endometrium may not reflect cell-specific transcription.…”

Actions of putative embryokines on development of the preimplantation bovine embryo to the blastocyst stage

“…It is likely, therefore, that CNP either affects the differentiation of Three of the molecules tested in the current study may participate in modification of embryonic development caused by specific inflammatory events in the endometrium. Both IL6 and IL8 are among the genes whose expression is increased in bovine endometrium associated with subclinical endometritis (Fagundes et al, 2019) and CNP is a constituent of bull seminal plasma (Chrisman et al, 1993). More research is required but evidence from this and other experiments would indicate that CNP and IL-6 would be facilitatory to embryo survival given the effects of CNP (this study) and IL-6 to increase ICM cell number and of IL-6 to activate JAK/STAT signaling in ICM this study).…”

“…C-Natriuretic peptide is also present in bull seminal plasma (Chrisman et al, 1993) and could conceivably be one of the mediators of seminal plasma effects on the endometrium or embryo (Bromfield, 2018). Interleukin-8 is one of the cytokines whose expression can be upregulated in females experiencing uterine inflammation or infectious disease (Fagundes et al, 2019;Helfrich et al, 2020) as well as exposure to sperm (Akthar et al, 2020). The primary objective of the current experiments was to evaluate actions of these molecules to increase the capacity of the bovine embryo to develop to the blastocyst stage.…”