脂多糖对体外培养奶牛子宫内膜上皮细胞子宫容受性因子表达的影响

doi:10.16431/j.cnki.1671-7236.2019.08.012

Abstract

Abstract:

The aim of this study was to establish an in vitro infection model of endometritis by inducing endometrial epithelial cells of dairy cows with lipopolysaccharide (LPS) and study the effects of endometritis on the relative expression of uterine receptive factors MUC-1, Wnt-7a, IFNAR1, IFNAR2, Integrin αvβ3 mRNA and protein in dairy cows.The mechanism of repeated sterility and low reproductive rate in dairy cows caused by endometritis were further clarified.Endometrial epithelial cells of dairy cows were isolated and cultured by tissue block method;The purity of the cells was determined by immunofluorescence;Endometrial epithelial cells were stimulated by LPS at different concentrations;The cell survival rate was measured by CCK8 at different time points;The secretion of inflammatory factors interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), IL-6 and IL-8 were detected by ELISA.Real-time PCR and Western blotting were used to detect the mRNA and protein expression of uterine receptive factors MUC-1, Wnt-7a, IFNAR1, IFNAR2, Integrin αvβ3 in endometrial infection model.The results showed that the number of bovine uterine epithelial cells purified by tissue block method was more, and the purity was higher by immunofluorescence keratin staining.CCK8 assay showed that compared with control group, LPS at 0, 5, 10, 50 μg/mL had no significant change in cell viability after 6, 12, 24 and 48 h (P>0.05).When LPS concentration was 100 μg/mL for 24 h, the cell survival rate was significantly lower than that of control group (P<0.05).The contents of IL-1β, TNF-α, IL-6 and IL-8 in cell culture supernatant were significantly higher than those in control and low concentration groups (P<0.05).Compared with control group, the relative expression of MUC-1 mRNA and protein in the model group extremely significantly increased (P<0.01), Wnt-7a mRNA and protein extremely significantly decreased (P<0.01).The mRNA expression of Integrin αvβ3, IFNAR1 and IFNAR2 extremely significantly decreased (P<0.01), and their protein expression were significantly decreased (P<0.05).The results showed that LPS with concentration of 100 μg/mL for 24 h could successfully construct the infection model of endometritis in vitro.The effects of endometritis on the expression of MUC-1, Wnt-7a, IFNAR1, IFNAR2, Integrin αvβ3 mRNA and protein might be the important reasons for repeated sterility and low reproductive rate of dairy cows.

Key words: dairy cow; endometrial epithelial cells; lipopolysaccharide (LPS); uterine receptive factor

CLC Number:

S823.9⁺1

FENG Yan, NI Hemin, GUO Yong, SHENG Xihui, QI Xiaolong, XING Kai, WANG Xiangguo. Effect of Lipopolysaccharide on Expression of Uterine Receptive Factors in Dairy Cow Endometrial Epithelial Cells Cultured in vitro[J]. China Animal Husbandry & Veterinary Medicine, 2019, 46(8): 2288-2295.

References 32

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Effect of Lipopolysaccharide on Expression of Uterine Receptive Factors in Dairy Cow Endometrial Epithelial Cells Cultured in vitro

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