Background

Neisseria gonorrhoeae (N.g) is Gram-negative bacteria and can lead to endometritis in female. Toll-like receptors regulate immune response in various diseases. However, the roles of TLR2 and TLR4 in. Neisseria gonorrhoeae-induced infection damage in human endometrial epithelia were investigated.

Methods

hEECs were infected with N.g (MOI 10 and 100) and cell viability and apoptosis were measured by CCK8 and flow cytometry assays in both infected groups with the uninfected normal hEECs as negative control. TLR2/TLR4 proteins were measured by ELISA method. Pro-inflammatory markers NLRP3, PGES (PGE2) and TNF-α were assessed by RT-qPCR (mRNA expression) and Elisa (protein concentrations). Transfection assays were performed to up- or down- regulate expression of TLR2 and TLR4 so as to study the functions of TLR2/TLR4 in. N.g-infected hEECs, followed by apoptosis and inflammation assessment. Similarly, we explored the interactions between TLR2/TLR4 and Nrf2/NF-κB/p65 by knocking down TLR2/TLR4 to detect the signaling and further regulating the signaling to evaluate TLR2/ TLR4, apoptosis and inflammation in cells.

Results

N.g suppressed cell viabilities and induced cell apoptosis and inflammation. TLR2/TLR4 downregulation inhibited the infection damage. Nrf2 was activated while NF-κB/p65 was depleted as TLR2/ TLR4 was knocked down. Activation of Nrf2 and inhibition of NF-κB resulted in decrease of TLR2/TLR4, which could retard apoptosis and inflammation induced by N.g infection.

Conclusion

TLR2/TLR4 depletion could alleviate the N.g-infected hEECs via Nrf2/NF-kB signaling, suggesting that TLR2/TLR4 inhibitors might serve as a treatment to reduce N.g infection in human endometrial epithelia.

中文翻译：

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抑制 TLR2/TLR4 通过 Nrf2 和 NF-Kβ 信号传导减轻人子宫内膜上皮细胞中淋病奈瑟菌感染的损伤。

背景

淋病奈瑟菌 (Ng) 是革兰氏阴性菌，可导致女性子宫内膜炎。Toll 样受体调节各种疾病的免疫反应。然而，研究了 TLR2 和 TLR4 在淋病奈瑟菌诱导的人类子宫内膜上皮感染损伤中的作用。

方法

hEEC 用 Ng（MOI 10 和 100）感染，并通过 CCK8 和流式细胞术测定在两个感染组中测量细胞活力和细胞凋亡，以未感染的正常 hEEC 作为阴性对照。通过ELISA方法测量TLR2/TLR4蛋白。促炎标志物 NLRP3、PGES (PGE2) 和 TNF-α 通过 RT-qPCR（mRNA 表达）和 Elisa（蛋白质浓度）进行评估。进行转染测定以上调或下调 TLR2 和 TLR4 的表达，以研究 TLR2/TLR4 在 Ng 感染的 hEEC 中的功能，然后进行细胞凋亡和炎症评估。同样，我们通过敲低 TLR2/TLR4 来检测信号并进一步调节信号以评估 TLR2/TLR4、细胞凋亡和炎症，从而探索了 TLR2/TLR4 和 Nrf2/NF-κB/p65 之间的相互作用。

结果

Ng 抑制细胞活力并诱导细胞凋亡和炎症。TLR2/TLR4 下调抑制感染损伤。Nrf2 被激活，而 NF-κB/p65 被耗尽，因为 TLR2/TLR4 被击倒。Nrf2 的激活和 NF-κB 的抑制导致 TLR2/TLR4 的减少，这可以延缓 Ng 感染诱导的细胞凋亡和炎症。

结论

TLR2/TLR4 耗竭可以通过 Nrf2/NF-kB 信号减轻 Ng 感染的 hEEC，表明 TLR2/TLR4 抑制剂可能作为一种治疗方法来减少人子宫内膜上皮细胞中的 Ng 感染。