AUTHOR=Lv Haimiao , Yan Chenbo , Deng Lixin , Peng Zhan , Yang Dexin , Hu Wenjv , Ding Xuefen , Tong Chao , Wang Xinzhuang 

TITLE=Role of MicroRNAs in Protective Effects of Forsythoside A Against Lipopolysaccharide-Induced Inflammation in Bovine Endometrial Stromal Cells

JOURNAL=Frontiers in Veterinary Science

VOLUME=Volume 8 - 2021

YEAR=2021

URL=https://www.frontiersin.org/journals/veterinary-science/articles/10.3389/fvets.2021.642913

DOI=10.3389/fvets.2021.642913

ISSN=2297-1769

ABSTRACT=Bovine endometrial stromal cells (bESCs) are exposed to a complex environment of
bacteria and viruses due to the rupture of epithelial cells after delivery. Inflammatory
responses are elicited by the activation of host pattern recognition receptors through
pathogen-related molecules such as lipopolysaccharides (LPS) on the cell membrane.
Forsythoside A (FTA) is a major active constituent of Forsythia suspensa (Thunb.)
Vahl. Forsythia suspensa is a flowering plant widely employed as a traditional
Chinese herbal medicine to treat various inflammatory diseases such as nephritis, eye2
swelling, scabies, ulcers, and mastitis; however, the molecular mechanisms
underlying its therapeutic effects on bovine endometritis are still unclear. The aim of
this study was to explore the role of miRNA and the mechanisms underlying the
protective activity of FTA on the inflammation of bovine endometrial stromal cells
induced by LPS. Based on previous research, we isolated and cultured bESCs in vitro
and categorised them into LPS and LPS+FTA groups with three replicates. Upon
reaching 80% confluence, the bESCs were treated with 0.5 µg/mL of LPS or 0.5
µg/mL of LPS + 100 µg/mL of FTA. We, then, performed high-throughput
sequencing (RNA-Seq) to investigate the effects of FTA on LPS-stimulated primary
bESCs and their underlying mechanisms. We identified 167 miRNAs to be
differentially expressed in the LPS groups; 72 miRNAs were up-regulated and 95
were down-regulated. Gene ontology enrichment analysis revealed that differentially
expressed microRNA (DEGs) were most enriched during the cellular metabolic
process; they were mostly located intracellularly and participated in protein, enzyme,
and ion binding. Kyoto Encyclopedia of Genes and Genomes pathway analysis
revealed that the DEGs were most enriched in the mitogen-activated protein kinase,
tumour necrosis factor, and Interleukin-17 signalling pathways. These results reveal
the complex molecular mechanism involved in the FTA and provide a basis for future
studies of the treatment of bovine endometritis with traditional Chinese medicine
monomer.