Abstract：
OBJECTIVE: Innate lymphoid cells (ILCs) are a class of newly discovered immunocytes. Group 1 ILCs (ILC1s) are identified in the decidua of humans and mice. High mobility group box 1 (HMGB1) is predicted to be one of the target genes of miR-142-3p, which is closely related to pregnancy-related diseases. Furthermore, miR-142-3p and HMGB1 are involved in regulating the NF-κB signaling pathway. This study aimed to examine the regulatory effect of miR-142-3p on ILC1s and the underlying mechanism involving HMGB1 and the NF-κB signaling pathway.
METHODS: Mouse models of normal pregnancy and abortion were constructed, and the alterations of ILC1s, miR-142-3p, ILC1 transcription factor (T-bet), and pro-inflammatory cytokines of ILC1s (TNF-α, IFN-γ and IL-2) were detected in mice from different groups. The targeting regulation of HMGB1 by miR-142-3p in ILC1s, and the expression of HMGB1 in normal pregnant mice and abortive mice were investigated. In addition, the regulatory effects of miR-142-3p and HMGB1 on ILC1s were detected in vitro by CCK-8, Annexin-V/PI, ELISA, and RT-PCR, respectively. Furthermore, changes of the NF-κB signaling pathway in ILC1s were examined in the different groups. For the in vivo studies, miR-142-3p-Agomir was injected in the uterus of abortive mice to evaluate the abortion rate and alterations of ILC1s at the maternal-fetal interface, and further detect the expression of HMGB1, pro-inflammatory cytokines, and the NF-κB signaling pathway.
RESULTS: The number of ILC1s was significantly increased, the level of HMGB1 was significantly upregulated, and that of miR-142-3p was considerably downregulated in the abortive mice as compared with the normal pregnant mice (all P<0.05). In addition, miR-142-3p was found to drastically inhibit the activation of the NF-κB signaling pathway (P<0.05). The number of ILC1s and the levels of pro-inflammatory cytokines were significantly downregulated and the activation of the NF-κB signaling pathway was inhibited in the miR-142-3p Agomir group (all P<0.05).
CONCLUSIONS: miR-142-3p can regulate ILC1s by targeting HMGB1 via the NF-κB signaling pathway, and attenuate the inflammation at the maternal-fetal interface in abortive mice.
METHODS: Mouse models of normal pregnancy and abortion were constructed, and the alterations of ILC1s, miR-142-3p, ILC1 transcription factor (T-bet), and pro-inflammatory cytokines of ILC1s (TNF-α, IFN-γ and IL-2) were detected in mice from different groups. The targeting regulation of HMGB1 by miR-142-3p in ILC1s, and the expression of HMGB1 in normal pregnant mice and abortive mice were investigated. In addition, the regulatory effects of miR-142-3p and HMGB1 on ILC1s were detected in vitro by CCK-8, Annexin-V/PI, ELISA, and RT-PCR, respectively. Furthermore, changes of the NF-κB signaling pathway in ILC1s were examined in the different groups. For the in vivo studies, miR-142-3p-Agomir was injected in the uterus of abortive mice to evaluate the abortion rate and alterations of ILC1s at the maternal-fetal interface, and further detect the expression of HMGB1, pro-inflammatory cytokines, and the NF-κB signaling pathway.
RESULTS: The number of ILC1s was significantly increased, the level of HMGB1 was significantly upregulated, and that of miR-142-3p was considerably downregulated in the abortive mice as compared with the normal pregnant mice (all P<0.05). In addition, miR-142-3p was found to drastically inhibit the activation of the NF-κB signaling pathway (P<0.05). The number of ILC1s and the levels of pro-inflammatory cytokines were significantly downregulated and the activation of the NF-κB signaling pathway was inhibited in the miR-142-3p Agomir group (all P<0.05).
CONCLUSIONS: miR-142-3p can regulate ILC1s by targeting HMGB1 via the NF-κB signaling pathway, and attenuate the inflammation at the maternal-fetal interface in abortive mice.
摘要：
目的：固有淋巴细胞（ILC）是一类新发现的免疫细胞。在人和小鼠的蜕膜中鉴定出第1组ILC(ILCls)。高迁移率族蛋白1（HMGB1）被预测为miR-142-3p的靶基因之一，这与妊娠相关疾病密切相关。此外，miR-142-3p和HMGB1参与调控NF-κB信号通路。本研究旨在探讨miR-142-3p对ILC1s的调控作用及其涉及HMGB1和NF-κB信号通路的潜在机制。
方法：构建正常妊娠和流产小鼠模型，以及ILC1的改变，miR-142-3p,ILC1转录因子(T-bet),和ILC1s的促炎细胞因子（TNF-α，在来自不同组的小鼠中检测到IFN-γ和IL-2）。miR-142-3p在ILC1s中对HMGB1的靶向调控,研究HMGB1在正常妊娠小鼠和流产小鼠中的表达。此外,CCK-8、Annexin-V/PI在体外检测miR-142-3p和HMGB1对ILC1s的调节作用,ELISA,和RT-PCR，分别。此外，在不同的组中检查了ILC1s中NF-κB信号通路的变化。对于体内研究，将miR-142-3p-Agomir注射到流产小鼠的子宫中，以评估母胎界面的流产率和ILC1s的变化，并进一步检测HMGB1、促炎细胞因子的表达,和NF-κB信号通路。
结果：ILC1的数量显著增加，HMGB1水平显著上调,与正常妊娠小鼠相比，流产小鼠的miR-142-3p明显下调（均P<0.05）。此外,miR-142-3p可显著抑制NF-κB信号通路的激活(P<0.05)。miR-142-3pAgomir组ILC1s数量和促炎细胞因子水平显著下调，NF-κB信号通路的激活受到抑制（均P<0.05）。
结论：miR-142-3p可通过NF-κB信号通路靶向HMGB1调控ILC1s,并减轻流产小鼠母胎界面的炎症。
方法：构建正常妊娠和流产小鼠模型，以及ILC1的改变，miR-142-3p,ILC1转录因子(T-bet),和ILC1s的促炎细胞因子（TNF-α，在来自不同组的小鼠中检测到IFN-γ和IL-2）。miR-142-3p在ILC1s中对HMGB1的靶向调控,研究HMGB1在正常妊娠小鼠和流产小鼠中的表达。此外,CCK-8、Annexin-V/PI在体外检测miR-142-3p和HMGB1对ILC1s的调节作用,ELISA,和RT-PCR，分别。此外，在不同的组中检查了ILC1s中NF-κB信号通路的变化。对于体内研究，将miR-142-3p-Agomir注射到流产小鼠的子宫中，以评估母胎界面的流产率和ILC1s的变化，并进一步检测HMGB1、促炎细胞因子的表达,和NF-κB信号通路。
结果：ILC1的数量显著增加，HMGB1水平显著上调,与正常妊娠小鼠相比，流产小鼠的miR-142-3p明显下调（均P<0.05）。此外,miR-142-3p可显著抑制NF-κB信号通路的激活(P<0.05)。miR-142-3pAgomir组ILC1s数量和促炎细胞因子水平显著下调，NF-κB信号通路的激活受到抑制（均P<0.05）。
结论：miR-142-3p可通过NF-κB信号通路靶向HMGB1调控ILC1s,并减轻流产小鼠母胎界面的炎症。
