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In Vitro Analysis of LPS-Induced miRNA Differences in Bovine Endometrial Cells and Study of Related Pathways
by
and
Xinmiao Li
1,2,†,
Zhihao Zhang
1,3,†,
Xiangnan Wang
1,
Ligang Lu
4,
Zijing Zhang
1,
Geyang Zhang
3,
Jia Min
3,
Qiaoting Shi
1,
Shijie Lyu
1,
Qiuxia Chu
1,
Xingshan Qi
5,
Huimin Li
6,
Yongzhen Huang
2,*
Eryao Wang
1,* 1
Institute of Animal Husbandry, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China
2
College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China
3
College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China
4
Bijie Academy of Agricultural Sciences, Bijie 551700, China
5
Bureau of Animal Husbandry of Biyang County, Biyang 463700, China
6
Agricultural Comprehensive Administrative Law Enforcement Detachment of Zhengzhou, Zhengzhou 450044, China
*
Authors to whom correspondence should be addressed.
†
Co-first author.
Animals 2024, 14(23), 3367; https://doi.org/10.3390/ani14233367
Submission received: 28 September 2024
/
Revised: 6 November 2024
/
Accepted: 15 November 2024
/
Published: 22 November 2024
(This article belongs to the Section Cattle)
Simple Summary
Lipopolysaccharide (LPS) is one of the main triggering factors for endometritis in cows. However, the specific pathogenesis of LPS-induced endometritis in dairy cows is not yet fully understood. The aim of this study is to establish an in vitro endometritis model using lipopolysaccharide-induced bovine endometrial epithelial (BEND) cells. BEND cells were treated with different concentrations and durations of LPS. Additionally, miRNA sequencing and target gene prediction were performed on both normal BEND cells and LPS-treated endometrial epithelial cells to screen for differentially expressed microRNAs. This study provides a theoretical basis for further exploring the pathogenesis of endometritis.
Abstract
Lipopolysaccharide (LPS) is one of the main factors inducing endometritis in dairy cows. However, the specific pathogenesis of LPS-induced endometritis in dairy cows is not fully understood. The objective of this study was to establish an in vitro endometritis model using LPS-induced bovine endometrial epithelial (BEND) cells. BEND cells were treated with LPS of different concentrations and times. The cell-counting kit-8 (CCK-8) was used to detect the cell survival rate after LPS treatment, and quantitative real-time PCR (RT-qPCR) was used to detect the expression of control group and LPS-treated group of inflammatory factors interleukin-1 beta (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF-α). The results showed that the survival rate of endometrial epithelial cells stimulated by 5 μg/mL LPS for 6 h was 75.13%, and the expression of inflammatory factors was significantly increased. Therefore, 5 μg/mL LPS for 6 h could be selected as a suitable model for the study of inflammation. In addition, miRNA sequencing and target gene prediction was performed on normal and LPS-treated BEND cells. Among twenty-one differentially expressed miRNAs, six miRNAs were selected and their expression levels were detected by RT-qPCR, which were consistent with the sequencing results. Twenty-one differentially expressed miRNAs collectively predicted 17,050 target genes. This study provides a theoretical basis for further investigation of the pathogenesis of endometritis.
Keywords:
LPS; endometritis; inflammatory models; miRNA