子痫前期(PE)是一种发病机制复杂的妊娠特异性综合征。本研究旨在使用缺氧/复氧(H/R)处理的HTR8/SVneo细胞模型探讨热休克蛋白B8(HSPB8)和c-Myc在滋养细胞功能障碍中的作用。使用基因表达综合数据库分析PE患者组织中的HSPB8表达。在H/R刺激的HTR8/SVneo细胞中检测到HSPB8表达后,通过用HSPB8特异性质粒转染该基因来过表达HSPB8。细胞计数试剂盒-8,伤口愈合和Transwell测定用于评估增殖,暴露于H/R条件的HTR8/SVneo细胞的迁移和侵袭。通过2,7-二氯二氢荧光素二乙酸染色确定活性氧(ROS)。5,5\',6,6'-四氯-1,1',应用3,3'-四乙基苯并咪唑碳碘化花青(JC-1)染色评估线粒体膜电位。使用相应的商业试剂盒检测丙二醛(MDA)和超氧化物歧化酶(SOD)水平。此外,通过末端脱氧核苷酸转移酶dUTP缺口末端标记(TUNEL)染色评估细胞凋亡的诱导。此外,Biogrid数据库预测HSPB8与c-Myc绑定,并且使用共免疫沉淀(Co-IP)测定来验证这种相互作用。随后,c-Myc表达被沉默以在暴露于H/R条件和上调HSPB8表达的HTR8/SVneo细胞中进行拯救实验。值得注意的是,PE组织和H/R刺激的HTR8/SVneo细胞中HSPB8表达降低。HSPB8强制表达促进增殖,HTR8/SVneo细胞的迁移和侵袭。此外,H/R导致ROS和MDA水平增加以及TUNEL染色和聚集的JC-1荧光和SOD活性水平降低,HSPB8过表达后恢复。Co-IP证实了HSPB8和c-Myc之间的相互作用。此外,c-Myc表达的敲低损害了HSPB8上调对H/R诱导后滋养细胞功能障碍的影响。总的来说,数据表明,HSPB8可以通过与c-Myc结合来改善线粒体氧化应激,从而减轻滋养细胞功能障碍。这些发现可能为PE的发病机制提供新的见解,并强调HSPB8/c-Myc在未来PE的预防和治疗中的作用。
Preeclampsia (PE) is a pregnancy-specific syndrome with complex pathogenesis. The present study aimed to explore the role of heat shock protein B8 (
HSPB8) and c-Myc in trophoblast cell dysfunction using a hypoxia/reoxygenation (H/R)-treated HTR8/SVneo cell model. HSPB8 expression in tissues of patients with PE was analyzed using the Gene Expression Omnibus database. Following detection of
HSPB8 expression in H/R-stimulated HTR8/SVneo cells,
HSPB8 was overexpressed by transfection of the gene with a
HSPB8-specific plasmid. Cell Counting Kit-8, wound healing and Transwell assays were used to evaluate the proliferation, migration and invasion of HTR8/SVneo cells exposed to H/R conditions. Reactive oxygen species (ROS) were determined by 2,7-dichlorodihydrofluorescein diacetate staining. 5,5\',6,6\'-tetrachloro-1,1\',3,3\'-tetraethylbenzimidazolocarbo-cyanine iodide (JC-1) staining was applied to assess mitochondrial membrane potential. Malondialdehyde (MDA) and superoxide dismutase (SOD) levels were detected using the corresponding commercial kits. In addition, the induction of apoptosis was assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Moreover, the Biogrid database predicted that HSPB8 was bound to c-Myc, and a co-immunoprecipitation (Co-IP) assay was used to verify this interaction. Subsequently, c-Myc expression was silenced to conduct rescue experiments in HTR8/SVneo cells exposed to H/R conditions and upregulated HSPB8 expression. Notably, reduced
HSPB8 expression was noted in PE tissues and H/R-stimulated HTR8/SVneo cells. HSPB8 enforced expression promoted the proliferation, migration and invasion of HTR8/SVneo cells. Moreover, H/R caused an increase in ROS and MDA levels as well as in TUNEL staining and a decrease in aggregated JC-1 fluorescence and SOD activity levels, which were restored following HSPB8 overexpression. Co-IP confirmed the interaction between
HSPB8 and c-Myc. Moreover, knockdown of c-Myc expression compromised the effects of HSPB8 upregulation on trophoblast cell dysfunction following induction of H/R. Collectively, the data indicated that HSPB8 could improve mitochondrial oxidative stress by binding to c-Myc to alleviate trophoblast cell dysfunction. The findings may provide new insights into the pathogenesis of PE and highlight the role of HSPB8/c-Myc in the prevention and treatment of PE in the future.