Aucubin ameliorates the LPS-induced inflammatory response in bovine endometrial epithelial cells by inhibiting NF-κB and activating the Keap1/Nrf2 signalling pathway
Aucubin ameliorates the LPS-induced inflammatory response in bovine endometrial epithelial cells by inhibiting NF-κB and activating the Keap1/Nrf2 signalling pathway
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DOI：
10.1111/rda.13939
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发表时间：
2021-04-27
影响因子：
1.7
通讯作者：
Fu, Kaiqiang
中科院分区：
文献类型：
--
作者：
Gao, Feng;Li, Huatao;Fu, Kaiqiang
奶牛在怀孕后容易受到病原性细菌感染，导致子宫内膜炎症。桃叶珊瑚苷（Au）已被证明具有高效的抗炎活性，但其预防奶牛乳房炎的能力尚不清楚。因此，本研究的目的是评估Au对LPS诱导的牛子宫内膜上皮细胞（BEECs）炎症反应的保护作用。用Au（10、20和50 μ M）预处理BEEC 6 h后，用LPS刺激细胞3 h，随后通过流式细胞术分析BEEC细胞凋亡，通过qRT-PCR检测促炎细胞因子mRNA的表达，并通过免疫印迹和免疫荧光分析NF-κ B和Keap 1/Nrf 2信号传导的变化。结果表明，Au可降低BEEC中TNF-α、IL-1 β、IL-6、考克斯-2和iNOSmRNA的表达，减少细胞凋亡。此外，Au显著降低NF-κ B p65和I κ B磷酸化水平，并抑制NF-κ B p65的核转位。Au还激活了Keap 1/Nrf 2通路，促进Nrf 2的核转移，并增加Keap 1、Nrf 2、HO-1和NQO 1的mRNA和蛋白水平。综上所述，这些结果表明，Au通过抑制NF-κ B和激活Keap 1/Nrf 2信号通路来改善LPS诱导的炎症反应，这对BEEC具有保护作用。
Cows are susceptible to pathogenic bacterial infection after pregnancy, leading to inflammation of the endometrium. Aucubin (AU) has been proven to exhibit highly effective anti-inflammatory activity, but its ability to protect against endometritis in dairy cows remains unclear. Therefore, the goal of the present study was to evaluate the protective effect of AU on the LPS-induced inflammatory response of bovine endometrial epithelial cells (BEECs). After pre-treating BEECs with AU (10, 20 and 50 mu M) for 6 hr, the cells were stimulated with LPS for 3 hr. Subsequently, BEECs apoptosis was analysed by flow cytometry, the expression of pro-inflammatory cytokine mRNA was detected by qRT-PCR, and changes in NF-kappa B and Keap1/Nrf2 signalling were analysed by western blotting and immunofluorescence analyses. The results showed that AU can reduce TNF-alpha, IL-1 beta, IL-6, COX-2 and iNOS mRNA expression in BEECs and reduce cell apoptosis. Furthermore, AU significantly reduced the level of NF-kappa B p65 and I kappa B phosphorylation and inhibited the nuclear translocation of NF-kappa B p65. AU also activated the Keap1/Nrf2 pathway, promoting the nuclear transfer of Nrf2 and increasing Keap1, Nrf2, HO-1 and NQO1 mRNA and protein levels. Taken together, these results indicate that AU ameliorates the LPS-induced inflammatory response by inhibiting NF-kappa B and activating the Keap1/Nrf2 signalling pathway, which has a protective effect on BEECs.