miR-15a、miR-466b-3p、miR-328-3p和miR-144-5p对硝酸钕诱导小于胎龄儿绒毛膜细胞凋亡的调控作用  

Regulatory effects of miR-15a,miR-466b-3p,miR-328-3p,and miR-144-5p on neodymium nitrate induced proliferation and apoptosis of chorionic villus in small for gestational age infants

作　　者：张宏波 郭婧[2,3] 张琦 霍宇婷 程霞霞 邰大鹏 乔瑞[1] ZHANG Hongbo;GUO Jing;ZHANG Qi;HUO Yuting;CHENG Xiaxia;TAI Dapeng;QIAO Rui(Baotou Medical College,Baotou,014040,China;SINOPHARM North Hospital of Baotou;Kundulun District Hospital)

机构地区：[1]包头医学院,内蒙古包头014040 [2]包头市国药北方医院 [3]包头市昆都仑区医院

出　　处：《包头医学院学报》2024年第10期6-11,86,共7页Journal of Baotou Medical College

基　　金：国家自然科学基金项目(82060600);内蒙古自治区卫生健康科研计划项目(20221391)。

摘　　要：目的:探究硝酸钕[Nd(NO 3)3]对小于胎龄儿(small for gestational age,SGA)绒毛膜细胞凋亡及标志性microRNA表达变化的影响,揭示钕致SGA的潜在分子作用机制。方法:选取人绒毛膜细胞系(JEG-3),通过CCK-8确定Nd(NO 3)3最佳染毒剂量、时间;流式细胞术检测细胞凋亡情况;提取对照组(0μg/mL)、实验组[低、中、高剂量组(0.2、2、20μg/mL)]细胞的miRNA进行qPCR,检测miR-15a、miR-466b-3p、miR-328-3p、miR-144-5p的表达水平。转染miR-15a、miR-144-5p抑制剂后检测细胞转染情况,再次进行CCK-8、流式细胞术验证细胞凋亡情况。结果:CCK-8结果显示,中剂量组细胞存活率下降至75.53%,流式细胞检测同样发现在中剂量组出现了更多凋亡。对JEG-3细胞染毒24 h提取细胞miRNA检测其miR-15a、miR-466b-3p、miR-328-3p、miR-144-5p表达水平,与对照组相比均出现不同程度上调。JEG-3细胞系转染上述microRNA抑制剂后,CCK-8中剂量组miR-144-5p、miR-15a抑制剂转染组其存活率由原75.53%回升至76.71%、83.03%,而凋亡水平则由原18.70%下降至18.44%、15.34%。结论:硝酸钕对JEG-3毒性影响呈现剂量依赖性,miR-15a、miR-466b-3p、miR-328-3p、miR-144-5p表达水平出现上调,激活下游信号通路调节细胞凋亡,对硝酸钕引起的细胞应激反应具有重要影响,为探究SGA的分子作用机制提供了新的研究方向。Objective:To investigate the effects of neodymium nitrate[Nd(NO 3)3]on chorionic cell apoptosis and the changes of microRNA expression in small for gestational age(SGA)infants,aiming to explore the molecular mechanisms of neodymium inducing SGA.Methods:The human trophoblast cell line(JEG-3)was selected.The optimal dose and time of Nd(NO 3)3 exposure were determined by CCK-8 assay.Apoptosis was measured using flow cytometry.MicroRNA(miRNA)was extracted from the control group(0μg/mL)and experimental group treated with low,medium,and high doses(0.2,2,20μg/mL)of Nd(NO 3)3.The expression levels of miR-15a,miR-466b-3p,miR-328-3p,and miR-144-5p were measured by qPCR.After transfection with miR-15a and miR-144-5p inhibitors,transfection efficiency was evaluated,and CCK-8 and flow cytometry assays were repeated to confirm their effects on apoptosis.Results:The CCK-8 assay showed that cell viability in the medium-dose group decreased to 75.53%,and Flow cytometry also indicated increased apoptosis in this group.After 24 hours of Nd(NO 3)3 exposure,qPCR analysis showed that miR-15a,miR-466b-3p,miR-328-3p,and miR-144-5p were upregulated compared to the control group.After JEG-3 cell line transfected with the above microRNA inhibitors,the survival rate of miR-144-5p and miR-15a inhibitor transfected group in CCK-8 medium-dose group increased from 75.53%to 76.71%and 83.03%,respectively,while the apoptosis level decreased from 18.70%to 18.44%and 15.34%respectively.Conclusion:The toxic effect of[Nd(NO 3)3]on cells is dose-dependent.With increased expression levels of miR-15a,miR-466b-3p,miR-328-3p,and miR-144-5p,downstream signaling pathway is activated and apoptosis mechanism is regulated,which plays an important role in cell stress response induced by(Nd(NO 3)3).This study provides a new research direction for exploring the molecular mechanism of SGA.

关 键 词：稀土元素钕 小于胎龄儿 胎盘 MIRNA表达 凋亡 

分 类 号：R714[医药卫生—妇产科学]