Involvement of interferon-tau in the induction of apoptotic, pyroptotic, and autophagic cell death-related signaling pathways in the bovine uterine endometrium during early pregnancy

Abstract

Interferon-tau (IFNT), a type I interferon (IFN), is known as pregnancy recognition signaling molecule secreted from the ruminant conceptus during the preimplantation period. Type I IFNs, such as IFN-alpha and IFN-beta, are known to activate cell-death pathways as well as induce apoptosis. In cows, induction of apoptosis with DNA fragmentation is induced by IFNT in cultured bovine endometrial epithelial cells. However, the status of cell-death pathways in the bovine endometrium during the preimplantation period still remains unclear. In the present study, we investigated the different cell-death pathways, including apoptosis, pyroptosis, and autophagy, in uterine tissue obtained from pregnant cows and in vitro cultured endometrial epithelial cells with IFNT stimulation. The expression of CASP7, 8, and FADD (apoptosis-related genes) was significantly higher in pregnant day 18 uterine tissue in comparison to non-pregnant day 18 tissue. The expression of CASP4, 11, and NLRP3 (pyroptosis-related genes) was significantly higher in the pregnant uterus in comparison to non-pregnant uterus. In contrast, autophagy-related genes were not affected by pregnancy. We also investigated the effect of IFNT on the expression of cell-death pathway-related genes, as well as DNA fragmentation in cultured endometrial epithelial cells. Similar to its effects in pregnant uterine tissue, IFNT affected the increase of apoptosis-related (CASP8) and pyroptosis-related genes (CASP11), but did not affect autophagy-related gene expression. IFNT also increased γH2AX-positive cells, which is a marker of DNA fragmentation. These results suggest that apoptosis- and pyroptosis-related genes are induced by IFNT in the pregnant bovine endometrial epithelial cells.

Heat stress induces oxidative stress and activates the KEAP1-NFE2L2-ARE pathway in bovine endometrial epithelial cells

“…Non-pregnant healthy cow uteri were obtained from a local slaughterhouse (Hokkaido Hayakita Meat Inspection Center, Japan). The isolation and culture of bovine endometrial epithelial cells from bovine endometrial tissues have been described previously [39,40]. The epithelial cells were cultured in Dulbecco's Modified Eagle's medium (high glucose) (DMEM; Wako, Osaka, Japan) supplemented with 5 % (v/v) fetal bovine serum (FBS; ICN Bio-Source International, Camarillo, CA, USA) and Antibiotic-Antimycotic solution (Thermo Fisher Scientific, Waltham, MA, USA) at 38.5 °C with 5% CO2, and were used within five passages.…”

“…listed in Table 1. The thermal cycling conditions were as follows: 1 cycle at 95 °C for 30 s, followed by 50 cycles at 95 °C for 10 s, 60 °C for 15 s, and 72 °C for 30 s. Relative mRNA abundance was calculated based on the expression levels of bovine H2AFZ, which was used as a reference gene as it show stable expression in bovine endometrium [39,40,42], and was relatively stable under heat stress conditions (our preliminary observations). Each run was completed with a melting curve analysis to confirm the specificity of amplification and the absence of primer dimer formation.…”

Sirt6-induced autophagy restricted TREM-1-mediated pyroptosis in ox-LDL-treated endothelial cells: relevance to prognostication of patients with acute myocardial infarction

“…For example, Atg7 deficiency intensifies inflammation and pyroptosis in Pseudomonas sepsis 34 and INF-mediated endometrial epithelial cells 35 . In contrast, autophagy deficiency using 3-MA in benzoapyrene-induced HL-7702 cells may effectively dampen pyroptosis 36 . Interestingly, autophagy restricts pyroptosis in cardiovascular diseases.…”

Modulation of Apoptosis by Bovine Gammaherpesvirus 4 Infection in Bovine Endometrial Cells and the Possible Role of LPS in This Process

“…The measurement of its expression level was conducted in accordance with the methodology described by Romeo et al in 2022 [19]. Specific primers for the BcL-2, Bax, Caspase 3 and GAPDH genes were used [26]. All samples were amplified in triplicate and the qPCR products were expressed as cycle threshold (Ct) values using the software StepOnePlus ® , version 2.0) (Applied Biosystems, Foster City, CA, USA).…”