Decreasing of S100A4 in bovine endometritis in vivo and in vitro

Li, Yajuan; Wang, Dongsheng; Wu, Xiaohu; He, Bin; Cheng, Zhangrui; Szenci, O.; Song, Pengjie; Shao, Dan; Zhang, Shidong; Yan, Zuoting

doi:10.1016/j.theriogenology.2020.05.015

Cited by 8 publications

(9 citation statements)

References 18 publications

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“…Bovine endometrial epithelial cells (BEECs) were isolated from a healthy uterus of a six-month-old dairy cow by our laboratory (35) and were cultured in DMEM supplemented with 10% FBS at 37°C in a humidified atmosphere with 5% CO 2 . LPS (1 mg) was diluted into 0, 3, 10, and 30 µg/ml by DMEM without FBS.…”

Section: Cell Culturementioning

confidence: 99%

LPS Mediates Bovine Endometrial Epithelial Cell Pyroptosis Directly Through Both NLRP3 Classical and Non-Classical Inflammasome Pathways

Shen

et al. 2021

Front. Immunol.

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As a highly inflammatory form of programmed cell death, pyroptosis is triggered by pro-inflammatory signals and associated with inflammation. It is characterized by cell swelling and large bubbles emerging from the plasma membrane, which release cytokines during inflammation. Compared with other types of cell death, pyroptosis has a distinct morphology and mechanism and involves special inflammasome cascade pathways. However, the inflammasome mechanism through which endometrial epithelial cell pyroptosis occurs in LPS-mediated inflammation remains unclear. We confirmed that there was an increased mRNA and protein expression of the IL-6, TNF-α, IL-1β, IL-18 cytokines, the inflammasome molecules NLRP3, CASPASE-1, CASPASE-4, and GSDMD in LPS-induced primary bovine endometrial epithelial cells (BEECs) in an in vitro established inflammatory model using ELISA, real-time PCR (RT-PCR), vector construction and transfection, and Western blotting. Scanning electron microscopy and lactate dehydrogenase (LDH) activity assays revealed induced cell membrane rupture, which is the main characteristic of pyroptosis. In conclusion, the cytolytic substrate GSDMD’s cleavage by caspase-1 or caspase-4 through the NLRP3 classical and non-classical inflammasome pathways, GSDMD N-terminus bind to the plasma membrane to form pores and release IL -18, IL-1β cause cell death during LPS induced BEECs inflammation.

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Section: Cell Culturementioning

confidence: 99%

LPS Mediates Bovine Endometrial Epithelial Cell Pyroptosis Directly Through Both NLRP3 Classical and Non-Classical Inflammasome Pathways

Shen

et al. 2021

Front. Immunol.

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“…It has been widely accepted that the overproduction of pro-inflammatory mediators, including TNF-α, IL-6, IL-8, IL-1β, can be harmful and result in various inflammatory diseases. 41,43 Previous studies showed that treatment of BEECs with ultrapure LPS led to an inflammatory response characterized by increased IL-6, IL-8, IL-1β, and TNF-α, mRNA expression and concentration. 22,23,[49][50][51] Similarly, our results showed upregulated mRNA expression of IL-6, IL-8, IL-1β, and TNF-α after LPS treatment.…”

Section: Discussionmentioning

confidence: 99%

“…Our laboratory isolated bovine endometrial epithelial cells (BEECs) from healthy dairy cows’ uterine tissues. 43 Briefly, the uteri are brought to the laboratory in a sterile PBS(P1020. Solarbio, Beijing, China) (pH 7.2) containing penicillin (100 mg/mL) and streptomycin (100 U/mL) from a local slaughter slab.…”

Section: Methodsmentioning

confidence: 99%

MicroRNA miR-24-3p Mediates the Negative Regulation of Lipopolysaccharide-Induced Endometrial Inflammatory Response by Targeting TNF Receptor-Associated Factor 6 (TRAF6)

Oladejo¹,

Li²,

Imam³

et al. 2022

JIR

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Purpose Endometritis is a female reproductive disease that affects the cattle industries development and microRNAs (miRNAs) play a pivotal role and critical regulators of the innate immune response in varieties of diseases. The present study intends to investigate the regulatory role of miR‐24-3p in the innate immune response involved in endometritis and evaluate its therapeutic potential. Methods Whole mice uteri and bovine endometrial epithelial cells (BEECs) were separately stimulated with LPS. The BEECs were also transfected with miR-24-3p mimic and negative control; si TRAF6 and siNC; pcDNA3.1 empty and pcDNA3.1(+) TRAF6 separately with LPS stimulation. The expression levels of miR‐24-3p and TRAF6 were measured via quantitative real‐time polymerase chain reaction (qRT-PCR) and Western blot, respectively. LPS‐induced inflammatory response assessed by inflammatory cytokines secretion and expression via ELISA and qRT-PCR. Bioinformatics analysis and luciferase reporter assay validated the interaction between miR‐24-3p and TRAF6 . The activation of the NF‐ĸB/MAPK pathway and p65 phosphorylation was investigated by Western blot and immunofluorescence assay, respectively. Results The expression of miR‐24-3p was decreased, and TRAF6 was elevated with an increased level of pro-inflammatory cytokines in LPS‐treated BEECs and mice uterus. The overexpression of miR‐24-3p suppressed LPS‐induced secretion of inflammatory cytokines (IL‐1β, IL‐6, IL-8 and TNF-α) and deactivation of NF‐ĸB/MAPK pathways. The downregulation of TRAF6 inhibited LPS‐induced inflammatory response in BEECs. TRAF6 is validated as a target of miR‐24-3p, and miR‐24-3p reversed the overexpression of cloned TRAF6 on inflammation response in BEECs. Conclusion Our findings demonstrate that the overexpression of miR‐24-3p attenuates endometrial inflammation and the expression of pro-inflammatory mediators via suppressing TRAF6 . Therefore, modulating the pathogenesis of endometritis and possibly, a therapeutic potential against endometritis.

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“…Uterus is normally sterile throughout pregnancy and exposed to many bacteria during post-partum period. Infection of endometrium with Gram-negative bacteria is the major pathogenesis of endometritis in cattle ( 22 ). Acting as parts of the innate host defense, cathelicidin is directly involved in pathogen clearing to providing a first line of defense against invading microbes ( 23 ).…”

Section: Discussionmentioning

confidence: 99%

Expression Pattern of Cathelicidins in Dairy Cows During Endometritis and Role of Bovine Endometrial Epithelial Cells in Production of Cathelicidins

Yang

et al. 2021

Front. Vet. Sci.

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Endometritis is a common bacterial disease of dairy cows. Cathelicidins are host-defense peptides that play important roles in clearance of bacteria. However, the expression pattern of these peptides during endometritis is still unclear. We hypothesize that the levels of bovine cathelicidins increased during endometritis. This study was to investigate the changes of bovine cathelicidins during endometritis. Forty-four post-partum cows (28–35 days after calving) involved in this study were grouped according to the character of vaginal discharge (VD) into three groups. These were (1) cows with clear fluid (n = 8, healthy cows group, N); (2) cows with VD containing <50% off-white mucopurulent material (n = 20, moderate endometritis cows, M); (3) cows with VD containing > 50% yellow or white purulent material (n = 16, severe endometritis cows, S). The blood, VD, and endometrial biopsies samples were collected from each cow to assess the levels of cathelicidin 1–7. Furthermore, bovine endometrial epithelial cells (BEECs) were stimulated with different concentration of Escherichia coli (2 × 106 and 2 × 107 CFU/mL) to detect the cellular source of cathelicidins. Quantitative real-time PCR (RT-qPCR) was used to detect the relative mRNA expression of cathelicidins, and enzyme-linked immune sorbent assay (ELISA) method were used to measure the protein levels. The mRNA and protein levels of cathelicidin 1–7 significantly increased during bovine endometritis (both moderate and severe endometritis), while samples from severe cases showed lower levels of cathelicidins compared to moderate cases. BEECs can express cathelicidin 1–7, and E. coli triggered the release of these proteins. High concentration of E. coli decreased the mRNA and protein levels of cathelicidins. Taken together, our results supported that cathelicidins are released as host defense molecules against the bacteria during bovine endometritis, and BEECs play an active role in expression and production of cathelicidins.

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Decreasing of S100A4 in bovine endometritis in vivo and in vitro

Cited by 8 publications

References 18 publications

LPS Mediates Bovine Endometrial Epithelial Cell Pyroptosis Directly Through Both NLRP3 Classical and Non-Classical Inflammasome Pathways

LPS Mediates Bovine Endometrial Epithelial Cell Pyroptosis Directly Through Both NLRP3 Classical and Non-Classical Inflammasome Pathways

MicroRNA miR-24-3p Mediates the Negative Regulation of Lipopolysaccharide-Induced Endometrial Inflammatory Response by Targeting TNF Receptor-Associated Factor 6 (TRAF6)

Expression Pattern of Cathelicidins in Dairy Cows During Endometritis and Role of Bovine Endometrial Epithelial Cells in Production of Cathelicidins

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