Abstract
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.
Highlights
In recent years, the incidence of uterine disease in dairy cows has shown an increasing trend, and has become one of the most important factors affecting their reproductive efficiency [1]
Based on the above information, this study focused on the inhibitory effects of bAD-mesenchymal stem cell (MSC) on LPS induced inflammation in bovine endometrial epithelial cells along with the underlying molecular mechanisms in vitro
The results of immunofluorescence microscopy and flow cytometry assays showed that isolated cells were strongly stained with CD73 and CD44 antibodies, the positive surface markers for bAD-MSCs, but showed a low binding affinity to CD45 and CD34
Summary
The incidence of uterine disease in dairy cows has shown an increasing trend, and has become one of the most important factors affecting their reproductive efficiency [1]. Endometritis is a type of refractory uterine disease, which is mainly caused by pathogenic microorganisms (such as Escherichia coli and Streptococcus) infecting the endometrium during or after parturition [3]. Endometritis is one of the most common reasons for infertility in dairy cows and causes huge annual losses to the dairy industry. A survey of the literature shows that in the United States uterine inflammation can cause losses of around US$171.9 ± 47.88 in first lactation primiparous heifers, while this figure is around US$262.65 ± 56.15 per lactation in multiparous cows [4]. There is an urgent need to find new and effective treatments for endometritis in dairy cows
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