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

2021 ◽  
Vol 8 ◽  
Author(s):  
Yajuan Li ◽  
Xiaoyu Ma ◽  
Jie Yang ◽  
Xiaohu Wu ◽  
Zuoting Yan ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Dairy Cows ◽  
Expression Pattern ◽  
Host Defense ◽  
Post Partum ◽  
Bacterial Disease ◽  
Host Defense Peptides ◽  
E Coli ◽  
Protein Levels ◽  
Endometrial Epithelial Cells

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.

scholarly journals Different effects of cortisol on pro-inflammatory gene expressions in LPS-, heat-killed E.coli-, or live E.coli-stimulated bovine endometrial epithelial cells

2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Luying Cui ◽  
Yali Wang ◽  
Heng Wang ◽  
Junsheng Dong ◽  
Zixiang Li ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Epithelial Cells ◽  
Dairy Cows ◽  
Bacterial Infections ◽  
Gene Expressions ◽  
Tlr4 Mrna ◽  
Tnf Α ◽  
Peripartum Period ◽  
Inflammatory Action ◽  
Endometrial Epithelial Cells

Abstract Background Bacterial infections are common in postpartum dairy cows. Cortisol level has been observed to increase in dairy cows during peripartum period, and is associated with the endometrial innate immunity against pathogens like E.coli. However, the mechanism underlying how cortisol regulates E.coli-induced inflammatory response in bovine endometrial epithelial cells (BEEC) remains elusive. Results Cortisol decreased the expressions of IL1β, IL6, TNF-α, IL8, and TLR4 mRNA in BEEC treated with LPS or heat-killed E.coli, but up-regulated these gene expressions in BEEC stimulated by live E.coli. Conclusion Cortisol exerted the anti-inflammatory action on LPS- or heat-killed E.coli-stimulated BEEC, but the pro-inflammatory action on live E.coli-induced BEEC.

Gene Expression Pattern Induced by Leptin in Human Endometrial Epithelial Cells

2005 ◽  
Vol 84 ◽  
pp. S435
Author(s):  
A. Cervero ◽  
J.A. Horcajadas ◽  
R. Catalano ◽  
A. Sharkey ◽  
A. Pellicer ◽  
...  
Keyword(s):  
Gene Expression ◽  
Epithelial Cells ◽  
Expression Pattern ◽  
Gene Expression Pattern ◽  
Endometrial Epithelial Cells

scholarly journals Different effects of cortisol on pro-inflammatory gene expressions of LPS-, heat-killed E.coli-, or live E.coli-stimulated bovine endometrial epithelial cells

2019 ◽  
Author(s):  
Luying Cui ◽  
Yali Wang ◽  
Heng Wang ◽  
Junsheng Dong ◽  
Zixiang Li ◽  
...  
Keyword(s):  
Innate Immunity ◽  
Epithelial Cells ◽  
Dairy Cows ◽  
Bacterial Infections ◽  
Gene Expressions ◽  
Tlr4 Mrna ◽  
Tnf Α ◽  
Peripartum Period ◽  
Anti Inflammation ◽  
Endometrial Epithelial Cells

Abstract Background: Bacterial infections are common in postpartum dairy cows. Cortisol level has been observed to increase in dairy cows during peripartum period, and is associated with the endometrial innate immunity against pathogen like E.coli . However, the mechanism underlying how cortisol regulates E.coli -induced inflammatory response in bovine endometrial epithelial cells (BEEC) remains elusive. Results: Cortisol decreased the expressions of IL1β, IL6, TNF-α, IL8, and TLR4 mRNA in BEEC treated with LPS or heat-killed E.coli , but up-regulated the these gene expressions in BEEC stimulated by live E.coli . Conclusion: Cortisol exerted the anti-inflammation action on LPS- or heat-killed E.coli -stimulated BEEC, but the pro-inflammation action on live E.coli -induced BEEC.

scholarly journals Progesterone Inhibits LPS-Induced Oxidative Stress Through Nrf2/Keap1 Pathway in Bovine Endometrial Epithelial Cells

2021 ◽  
Author(s):  
Luying Cui ◽  
Qi Zhang ◽  
Jiaqi Zhang ◽  
Heng Wang ◽  
Junsheng Dong ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Progesterone Receptor ◽  
Oxidative Damage ◽  
Antioxidative Effect ◽  
Antioxidant Genes ◽  
Uterine Infection ◽  
Protein Levels ◽  
Total Antioxidant ◽  
Nrf2 Protein ◽  
Endometrial Epithelial Cells

Abstract Background Postpartum uterine infection can lead to endometrial inflammation and oxidative damage. Progesterone makes the animal more susceptible to uterine infection. Progesterone has been proved to play an anti-inflammatory role in inhibiting uterine innate immunity, and to reduce tissue oxidative damage. But the effect of progesterone on the oxidative damage of bovine endometrium has not been reported. The purpose of this study was to explore the effect and mechanism of progesterone (1, 3, and 5 ng/mL) on oxidative damage in primary bovine endometrial epithelial cells (BEEC) induced by lipopolysaccharide (LPS) from Escherichia coli. Results Compared with the LPS group, there were decreases (P < 0.05) in the levels of reactive oxygen and malondialdehyde, and increases (P < 0.05) in the activities of superoxide dismutase and catalase, and total antioxidant capacity in the cotreatment groups of progesterone and LPS. The cotreatment of LPS and P4 upregulated (P < 0.05) the mRNA abundance of antioxidant genes and the key protein levels in Nrf2/Keap1 pathway, and promoted the Nrf2 protein to enter the nucleus. The use of progesterone receptor antagonist mifepristone reversed the antioxidative effect of progesterone. Conclusions Progesterone protects BEEC from LPS-induced oxidative damage by activating Nrf2/Keap1 pathway through progesterone receptor.

159 CHANGES IN PROTEIN EXPRESSION PROFILES IN BOVINE ENDOMETRIAL EPITHELIAL CELLS (bEEC) FOLLOWING E. COLI LIPOPOLYSACCHARIDE CHALLENGE

2015 ◽  
Vol 27 (1) ◽  
pp. 170 ◽  
Author(s):  
Y. Z. Guo ◽  
C. Piras ◽  
A. Soggiu ◽  
M. Chanrot ◽  
R. Båge ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Epithelial Cells ◽  
Protein Expression ◽  
Expression Profiles ◽  
Time Of Flight ◽  
Cell Number ◽  
Differentially Expressed ◽  
E Coli ◽  
Maldi Tof ◽  
Endometrial Epithelial Cells

E. coli is one of the most frequent bacteria involved in uterine diseases. Lipopolysaccharide (LPS) is a component of the outer membrane of Gram-negative bacteria involved in the pathogenic processes leading to postpartum metritis and endometritis in cattle. It also causes inflammation of the endometrium. Increase of cell proliferation by LPS is part of the inflammatory process and has been reported in human epithelial and immune cells (Martin et al. 2000 J. Immunol. 165, 139–147) and from bovine endometrial epithelial cells (bEEC) (Guo et al. 2014 Reprod. Fertil. Dev. 26, 165–166). The aim of this study was to investigate possible changes in protein expression in relation with the proliferative response of bEEC after challenge with E. coli-LPS. In vitro culture of bEEC was performed from 3 cows. On passage 5, bEEC from each individual were exposed to 0, 8, and 16 µg mL–1 LPS for 72 h. At time 0 and 72 h later, attached cells were counted and for each time and LPS dosage, cells were frozen for proteomic analyses. The variation of cells number over time was analysed by ANOVA (SAS 9.1, proc GLM; SAS Institute, Inc., Cary, NC, USA). All samples were analysed (every sample run in triplicate) by 2-D gel electrophoresis coupled to matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF)/time-of-flight (TOF) mass spectrometry (MS) and shotgun nLC-MS/MS analysis. As reported before, a significant increase in cell number was observed for cells treated with 8 µg mL–1 LPS (P ≤ 0.001), whereas changes in cell number were highly variable and nonsignificant for 16 µg mL–1 LPS. From each sample, ~800 proteins were visualised. Results from 2-D gel coupled to MALDI-TOF/TOF were very reproducible (same responses between individual cows) and revealed changes in protein profiles very much related (from P < 0.05 to P < 0.01) to proliferative phenotypes for seven proteins. From shotgun analysis, 27 proteins were found significantly differentially expressed (P < 0.05 to P < 0.01) following exposure to LPS (21 up-regulated and 6 down-regulated). Among the 21 found as up-regulated, 20 were differentially expressed both for the 8 and 16 µg mL–1 LPS, whereas 5 out of 6 were down-regulated for both dosages. Differentially expressed proteins were associated to cell proliferation, apoptosis, oxidative stress, regulation of histones, allergy, and general cell metabolism pathways. Candidate proteins need to be confirmed from larger series of individuals and relevant pathways further studied. Research was partially funded by RMUSTV.

scholarly journals Changes in protein expression profiles in bovine endometrial epithelial cells exposed to E. coli LPS challenge

2017 ◽  
Vol 13 (2) ◽  
pp. 392-405 ◽  
Author(s):  
Cristian Piras ◽  
Yongzhi Guo ◽  
Alessio Soggiu ◽  
Metasu Chanrot ◽  
Viviana Greco ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Protein Expression ◽  
Bacterial Infection ◽  
Physiological Response ◽  
Expression Profiles ◽  
E Coli ◽  
Lps Challenge ◽  
Protein Expression Profiles ◽  
Endometrial Epithelial Cells ◽  
In Pregnancy

Proteomics of the physiological response of bEECs to LPS challenge to unravel the possible implication of bacterial infection in pregnancy establishment.

scholarly journals Inhibitory Effect of Bovine Adipose-Derived Mesenchymal Stem Cells on Lipopolysaccharide Induced Inflammation of Endometrial Epithelial Cells in Dairy Cows

2021 ◽  
Vol 8 ◽  
Author(s):  
Wengeng Lu ◽  
Zheng-Mei Xu ◽  
Qing Liu ◽  
Nan-Nan Yu ◽  
Jia-Bin Yu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Epithelial Cells ◽  
Dairy Cows ◽  
Osteogenic Differentiation ◽  
Surface Marker ◽  
Economic Damage ◽  
Expression Levels ◽  
Bax Protein ◽  
Endometrial Epithelial Cells

Endometritis is a disease that affects reproductive health in dairy cows and causes serious economic damage to the dairy industry world-wide. Although in recent years, the application of mesenchymal stem cell (MSC) therapy for the treatment of inflammatory diseases has attracted much attention, there are few reports of the use of MSCs in dairy cows. In the present study, our objective was to explore the inhibitory effects of bovine adipose-derived mesenchymal stem cells (bAD-MSCs) on lipopolysaccharide (LPS) induced inflammation in bovine endometrial epithelial cells (bEECs) along with the potential underlying molecular mechanisms. We characterized isolated bAD-MSCs using cell surface marker staining and adipogenic/osteogenic differentiation, and analyzed them using immunofluorescence, flow cytometry (surface marker staining), and adipogenic and osteogenic differentiation. Furthermore, to understand the anti-inflammatory effects of bAD-MSCs on LPS induced bEEC inflammation, we used a bAD-MSC/bEEC co-culture system. The results showed that bAD-MSC treatments could significantly decrease LPS induced bEEC apoptosis and pro-inflammatory cytokine expression levels, such as interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). Furthermore, our results showed that bAD-MSC treatments could also significantly downregulate LPS induced p38, IkB-a, and JAK1 phosphorylation and Bax protein expression levels, which are closely related to inflammatory progress and cellular apoptosis in bEECs. Our findings demonstrate that bAD-MSCs play an inhibitory role in LPS induced bEEC inflammation and provide new insights for the clinical therapy of endometritis in dairy cows.

scholarly journals Conserved regulation of omptin proteases by the PhoPQ two-component regulatory system in Enterobacteriaceae

2020 ◽  
Author(s):  
Youn Hee Cho ◽  
Monir Riasad Fadle Aziz ◽  
Tanuja Sutradhar ◽  
Jasika Bashal ◽  
Veronica Cojocari ◽  
...  
Keyword(s):  
Binding Sites ◽  
Regulatory System ◽  
Enteric Bacteria ◽  
Gram Negative Bacteria ◽  
Citrobacter Rodentium ◽  
Host Defense Peptides ◽  
E Coli ◽  
Protein Levels ◽  
Regulatory Systems ◽  
Two Component

AbstractBacteria that colonize eukaryotic surfaces interact with numerous host-produced molecules that have antimicrobial activity. Bacteria have evolved numerous strategies to both detect and resist these molecules, and in gram-negative bacteria these include alterations of the cell surface lipopolysaccharide structure and/or charge and the production of proteases that can degrade these antimicrobial molecules. Many of the lipopolysaccharide alterations found in enteric bacteria are controlled by the PhoPQ and PmrAB two-component regulatory systems. Here, we show that omptin family proteases from Escherichia coli and Citrobacter rodentium are induced by growth in low Mg2+. We further show that deletion of PhoP eliminates omptin protease activity, transcriptional regulation and protein levels. We identify conserved PhoP-binding sites in the promoters of the E. coli omptin genes, ompT, ompP and arlC as well as in croP of Citrobacter rodentium and show that mutation of the putative PhoP-binding site in the ompT promoter abrogates PhoP-dependent expression. Finally, we show that despite the conserved PhoP-dependent regulation, each of the E. coli omptin proteins has differential activity toward a particular substrate, suggesting that each omptin may contribute to resistance to a particular repertoire of host-defense peptides, depending on the particular environment in which each evolved.

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