摘要
目的探究死亡受体3(death receptor 3,DR3)在胎盘滋养层细胞中的作用并解释相关机制。方法分别使用噻唑蓝比色法(methyl thiazolyl tetrazolium,MTT)、Transwell和流式细胞术分析评估人HTR-8/SVneo滋养层细胞的存活率、迁移、侵袭和凋亡。通过反转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-qPCR)分析DR3、磷脂酰肌醇3激酶(phosphatidylinositol 3-kinase,PI3K)和蛋白激酶B(protein kinase B,AKT)水平。利用酶联免疫吸附法检测DR3、p-PI3K、p-AKT、PI3K和AKT蛋白表达。结果与对照质粒相比,DR3质粒抑制了滋养层细胞的存活、迁移和侵袭能力,促进了细胞凋亡和活化Caspase8/Caspase8比值。此外,DR3质粒可降低p-PI3K和p-AKT的表达水平,使p-PI3K/PI3K的磷酸化比值由对照组的0.82降为0.19,p-AKT/AKT的磷酸化比值由对照组的0.80降为0.09。进一步实验表明,PI3K/AKT/CREB激活剂1可逆转DR3对上述细胞功能及PI3K/AKT信号通路的抑制作用。结论DR3通过抑制PI3K/AKT通路调节胎盘滋养层细胞的生理功能,对子痫前期进展产生作用。
Objective Investigate the role of death receptor 3(DR3)in placental trophoblast cells and elucidate the re-lated mechanisms.Methods The survival,migration,invasion and apoptosis of human HTR-8/SVneo trophoblastic cells were evaluated by methyl thiazolyl tetrazolium(MTT),Transwell and flow cytometry,respectively.The levels of DR3,phosphatidylinositol 3-kinase(PI3K)and protein kinase B(AKT)were analyzed by reverse transcription-polymerase chain reaction(RT-qPCR).The expression of DR3,p-PI3K,p-AKT,PI3K and AKT protein was detected by enzyme-linked immunosorbent assay.Results Compared with the control plasmid,DR3 plasmid inhibited the sur-vival,migration and invasion of trophoblast cells,promoted apoptosis and cleaved Caspase8/Caspase8 ratio.In addi-tion,DR3 plasmid could reduce the expression levels of p-PI3K and p-AKT.The phosphorylation ratio of p-PI3K/PI3K decreased from 0.82 to 0.19,and the phosphorylation ratio of p-AKT/AKT decreased from 0.80 to 0.09.Further experiments showed that PI3K/AKT/CREB activator 1 could reverse the inhibitory effect of DR3 on the above cell functions and PI3K/AKT signaling pathway.Conclusion DR3 modulates the physiological functions of placental tro-phoblast cells by inhibiting the PI3K/AKT pathway,thereby exerting an effect on the progression of preeclampsia.
作者
尹程
王佳慧
张瑜
张新平
沈彦祥
刘适
刘谡
赵薇
YIN Cheng;WANG Jiahui;ZHANG Yu;ZHANG Xinping;SHEN Yanxiang;LIU Shi;LIU Su;ZHAO Wei(Department of Obstetrics,The Third Affiliated Hospital of Qiqihar Medical University,Qiqihar 161000,Heilongjiang,China;Department of Gynecology,Section One,The Third Affiliated Hospital of Qiqihar Medical University,Qiqihar 161000,Heilongjiang,China;Department of Cardiology,Section Four,The Third Affiliated Hospital of Qiqihar Medical University,Qiqihar 161000,Heilongjiang,China;Central Laboratory,The Third Affiliated Hospital of Qiqihar Medical University,Qiqihar 161000,Heilongjiang,China)
出处
《系统医学》
2025年第12期34-39,共6页
Systems Medicine
基金
2022年度黑龙江省省属高等学校基本科研业务费科研项目(2022-KYYWF-0823)。
