【摘要】 目的 探讨血清微小RNA(miRNA)-146、脂多糖(LPS)和活性氧(ROS)在早期妊娠胚胎丢失(EPEL)患者中的水平及临床意义。方法 选取2021年3月至2023年3月广东省东莞市妇幼保健院收治的100例EPEL患者作为研究组，另外选取同期在广东省东莞市妇幼保健院就诊的100例正常妊娠女性作为对照组，检测并比较研究组和对照组血清指标水平。培养HTR-8/SVneo细胞，按照不同转染方式分成miRNA-146抑制剂组、miRNA-146模拟物组及NC组。比较miRNA-146抑制剂组、miRNA-146模拟物组Toll样受体4(TLR4)蛋白和核因子-κB(NF-κB)蛋白水平，比较NC组、miRNA-146模拟物组和miRNA-146抑制剂组miRNA-146水平及HTR-8/SVneo细胞的凋亡率，比较不同水平LPS处理的HTR-8/SVneo细胞生长活力。采用Pearson相关分析EPEL患者miRNA-146水平与ROS、LPS、白细胞介素(IL)-6、肿瘤坏死因子(TNF)-α、TLR4蛋白和NF-κB蛋白水平的相关性。结果 研究组LPS、ROS、IL-6和TNF-α水平高于对照组，且雌二醇、孕酮、miRNA-146水平低于对照组，差异均有统计学意义(P<0.05)。miRNA-146模拟物组miRNA-146水平高于miRNA-146抑制剂组和NC组，差异均有统计学意义(P<0.05)。miRNA-146模拟物组TLR4蛋白、NF-κB蛋白水平低于miRNA-146抑制剂组，差异均有统计学意义(P<0.05)。不同水平LPS(8、12、16、20μg/mL)处理的HTR-8/SVneo细胞生长活力低于20μg/mL LPS+miRNA-146模拟物处理的HTR-8/SVneo细胞生长活力，差异均有统计学意义(P<0.05)。miRNA-146抑制剂组HTR-8/SVneo细胞的凋亡率高于NC组和miRNA-146模拟物组，且NC组高于miRNA-146模拟物组，差异均有统计学意义(P<0.05)。Pearson相关分析结果显示，EPEL患者miRNA-146水平与ROS、LPS、IL-6、TNF-α、TLR4蛋白和NF-κB蛋白水平呈负相关(r=-0.737、-0.614、-0.712、-0.729、-0.739、-0.708,P<0.05)。结论 EPEL患者血清中miRNA-146水平明显下降，并与LPS、ROS、 IL-6、TNF-α的水平呈负相关，可能是EPEL发生的影响因素之一，有望为临床预测孕妇发生EPEL提供新思路。更多还原

【Abstract】 Objective To investigate the levels and clinical significance of serum microRNA(miRNA)-146,lipopolysaccharide(LPS) and reactive oxygen species(ROS) in patients with early pregnancy embryo loss(EPEL).Methods A total of 100 EPEL patients admitted to Dongguan Maternal and Child Health Hospital from March 2021 to March 2023 were selected as the study group, and 100 normal pregnant women admitted to Dongguan Maternal and Child Health Hospital during the same period were selected as the control group.The levels of serum indicators in the study group and the control group were detected and compared.HTR-8/SVneo cells were cultured and divided into miRNA-146 inhibitor group, miRNA-146 mimic group and NC group according to different transfection methods.The levels of Toll-like receptor 4(TLR4) protein and nuclear factor-κB(NF-κB) protein were compared between the miRNA-146 inhibitor group and the miRNA-146 mimic group.The miRNA-146 expession level and apoptosis rates of HTR-8/SVneo cells in NC group, miRNA-146 mimic group and miRNA-146 inhibitor group were compared, and the growth activity of HTR-8/SVneo cells treated with different levels of LPS was compared.Pearson correlation was used to analyze the correlation between miRNA-146 level and ROS,LPS,interleukin(IL)-6,tumor necrosis factor(TNF)-α,TLR4 protein and NF-κB protein levels in EPEL patients.Results The levels of LPS,ROS,IL-6 and TNF-α in the study group were higher than those in the control group, and the levels of estradiol, progesterone and miRNA-146 in the study group were lower than those in the control group, and the differences were statistically significant(P<0.05).The level of miRNA-146 in miRNA-146 mimic group was higher than that in miRNA-146 inhibitor group and NC group, and the differences were statistically significant(P<0.05).The levels of TLR4 protein and NF-κB protein in miRNA-146 mimics group were lower than those in miRNA-146 inhibitor group, and the differences were statistically significant(P<0.05).The growth activity of HTR-8/SVneo cells treated with different levels of LPS(8,12,16,20 μg/mL) were lower than those of HTR-8/SVneo cells treated with 20 μg/mL LPS+miRNA-146 mimic, respectively, and the differences were statistically significant(P<0.05).The apoptosis rate of HTR-8/SVneo cells in miRNA-146 inhibitor group was higher than that in NC group and miRNA-146 mimic group, and that in NC group was higher than that in miRNA-146 mimic group, and the differences were statistically significant(P<0.05).Pearson correlation analysis showed that the level of miRNA-146 was negatively correlated with the levels of ROS,LPS,IL-6,TNF-α,TLR4 protein and NF-κB protein in EPEL patients(r=-0.737,-0.614,-0.712,-0.729,-0.739,-0.708,P<0.05).Conclusion The serum miRNA-146 level in EPEL patients is significantly decreased and negatively correlated with LPS,ROS,IL-6 and TNF-α levels, which may be one of the influence factors of EPEL.It may provide a new idea for clinical prediction of EPEL in pregnant women.更多还原