IntroductionHyperglycemia is closely related to trophoblast dysfunction during pregnancy and results in suppressed invasion, migration, and pro‐inflammatory cell death of trophoblasts. Hyperglycemia is a dependent risk factor for gestational hypertension accompanied by decreased placental growth factor (PLGF), which is important for maternal and fetal development. However, there is currently a lack of evidence to support whether PLGF can alleviate trophoblast cell dysfunction caused by high blood sugar. Here, we aim to clarify the effect of hyperglycemia on trophoblast dysfunction and determine how PLGF affects this process.MethodsThe changes in placental tissue histomorphology from gestational diabetes mellitus (GDM) patients were compared with those of normal placentas. HTR8/SVneo cells were cultured in different amounts of glucose to examine cellular pyroptosis, migration, and invasion as well as PLGF levels. Furthermore, the levels of pyroptosis‐related proteins (NLRP3, pro‐caspase1, caspase1, IL‐1β, and Gasdermin D [GSDMD]) as well as autophagy‐related proteins (LC3‐II, Beclin1, and p62) were examined by Western blotting. The GFP‐mRFP‐LC3‐II system and transmission electron microscopy were used to detect mitophagy levels, and small interfering RNAs targeting BCL2 Interacting Protein 3 (siBNIP3) and PTEN‐induced kinase 1 (siPINK1) were used to determine the role of mitophagy in pyroptotic death of HTR‐8/SVneo cells.ResultsOur results show that hyperglycemia upregulates NLRP3, pro‐caspase1, caspase1, IL‐1β at the protein level in GDM patients. High glucose (HG, 25 mM) inhibits viability, invasion, and migration of trophoblast cells while suppressing superoxide dismutase levels and promoting malondialdehyde production, thus leading to a senescence associated beta‐gal‐positive cell burst. PLGF levels in nucleus and the cytosol are also inhibited by HG, whereas PLGF treatment inhibited pyroptosis‐related protein levels of NLRP3, pro‐caspase1, caspase1, IL‐1β, and GSDMD, Gasdermin D N‐terminal domain (GSDMD‐N). HG‐induced mitochondrial dysfunction and BNIP3 and PINK1/Parkin expression. Knocking down BINP3 and PINK1 abolished the protective role of PLGF by preventing mitophagy.ConclusionPLGF inhibited hyperglycemia, while PLGF reversed hyperglycemic injury by promoting mitophagy via the BNIP3/PINK1/Parkin pathway. Altogether, these results suggest that PLGF may protect against trophoblast dysfunction in diabetes.

中文翻译：

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胎盘生长因子通过调节线粒体自噬减轻高血糖诱导的滋养层细胞焦亡


简介高血糖与妊娠期滋养细胞功能障碍密切相关，导致滋养细胞的侵袭、迁移和促炎细胞死亡受到抑制。高血糖是妊娠期高血压的一个独立危险因素，伴有胎盘生长因子（PLGF）减少，而胎盘生长因子对母体和胎儿的发育很重要。但目前缺乏证据支持PLGF是否可以缓解高血糖引起的滋养层细胞功能障碍。在此，我们的目的是阐明高血糖对滋养层功能障碍的影响，并确定PLGF如何影响这一过程。方法比较妊娠期糖尿病（GDM）患者与正常胎盘的胎盘组织形态学变化。 HTR8/SVneo 细胞在不同量的葡萄糖中培养，以检测细胞焦亡、迁移和侵袭以及 PLGF 水平。此外，Western blot检测了细胞焦亡相关蛋白（NLRP3、pro-caspase1、caspase1、IL-1β和Gasdermin D [GSDMD]）以及自噬相关蛋白（LC3-II、Beclin1和p62）的水平。印迹。使用GFP-mRFP-LC3-II系统和透射电子显微镜检测线粒体自噬水平，并使用靶向BCL2相互作用蛋白3（siBNIP3）和PTEN诱导激酶1（siPINK1）的小干扰RNA来确定线粒体自噬在线粒体自噬中的作用。 HTR-8/SVneo 细胞焦亡。结果我们的结果表明，高血糖在 GDM 患者的蛋白质水平上调 NLRP3、pro-caspase1、caspase1、IL-1β。 高葡萄糖（HG，25 mM）抑制滋养层细胞的活力、侵袭和迁移，同时抑制超氧化物歧化酶水平并促进丙二醛产生，从而导致与衰老相关的 β-gal 阳性细胞爆发。细胞核和细胞质中的 PLGF 水平也受到 HG 的抑制，而 PLGF 治疗抑制了 NLRP3、pro-caspase1、caspase1、IL-1β 和 GSDMD、Gasdermin DN 末端结构域 (GSDMD-N) 的焦亡相关蛋白水平。 HG 诱导的线粒体功能障碍以及 BNIP3 和 PINK1/Parkin 表达。敲低BINP3和PINK1可消除PLGF通过阻止线粒体自噬的保护作用。结论PLGF抑制高血糖，而PLGF通过BNIP3/PINK1/Parkin通路促进线粒体自噬而逆转高血糖损伤。总而言之，这些结果表明 PLGF 可以预防糖尿病中的滋养层功能障碍。