He Mengzhou, Guo Xijiao, Jia Jing, Zhang Jingyi, Zhou Xuan, Wei Lijie, Yu Jun, Wang Shaoshuai, Feng Ling
Department of Gynecology and Obstetrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, PR China.
Department of Gynecology and Obstetrics, Wuhan Maternal and Child Health Hospital, Wuhan, Hubei, PR China.
Placenta. 2023 Oct;142:46-55. doi: 10.1016/j.placenta.2023.08.070. Epub 2023 Aug 23.
We investigated the proinflammatory functions of endoplasmic reticulum stress and peroxisome proliferator-activated receptor α (PPARα) in the development of gestational diabetes mellitus (GDM) and their relationship in regulating inflammation in GDM.
This study was performed on placentas of normal pregnant women, women with GDM, and HTR8 cells. Transmission electron microscopy, immunohistochemistry, Western blot analysis, and RT-PCR were performed to analyze ERS and PPARα expression on both normal and GDM pregnancy placentas. ELISA was performed to analyze inflammatory biomarkers. To generate models of the GDM-like state, placentas of normal pregnancy were treated with LPS and polyinosinic-polycytidylic acid (poly [I:C]). TG, CHOP plasmid, and CHOP siRNA were assessed as to their regulation of HTR8 cells to discern the relationship between ERS and PPARα in regulating the inflammation associated with GDM.
ERS was elevated in GDM placentas, induced the secretion of IL-6 and TNF-α, and attenuated the expression of GLUT-4. PPARα was diminished in GDM placentas and inhibited the inflammatory responses via the NF-κB nuclear-transport process. 4-PBA reduced CHOP and augmented PPARα, and it decreased IL-6 and TNF-α in our GDM-like explant. However, with both 4-PBA and MK886 treatment, we noted no significant difference in CHOP expression. The level of PPARα was reduced, and that of NF-κB p65 in the nucleus was elevated with TG treatment in the HTR8/Svneo. Knockdown of CHOP increased PPARα and reduced NF-κB p65, while expression of PPARα declined, and that of NF-κB p65 rose with the application of CHOP when HTR8 cells were treated with TG.
ERS contributes to the pathophysiology of GDM in pregnancy via the CHOP-PPARα-NF-κB-signalling pathway by inducing aberrant activation of inflammation and insulin resistance.
我们研究了内质网应激和过氧化物酶体增殖物激活受体α(PPARα)在妊娠期糖尿病(GDM)发生发展中的促炎功能及其在调节GDM炎症中的关系。
本研究对正常孕妇、GDM孕妇的胎盘及HTR8细胞进行。采用透射电子显微镜、免疫组织化学、蛋白质免疫印迹分析和逆转录-聚合酶链反应(RT-PCR)分析正常和GDM妊娠胎盘内质网应激(ERS)和PPARα的表达。采用酶联免疫吸附测定(ELISA)分析炎症生物标志物。为建立类GDM状态模型,用脂多糖(LPS)和聚肌苷酸-聚胞苷酸(聚[I:C])处理正常妊娠胎盘。评估三酰甘油(TG)、CHOP质粒和CHOP小干扰RNA(siRNA)对HTR8细胞的调节作用,以明确ERS和PPARα在调节与GDM相关炎症中的关系。
GDM胎盘ERS升高,诱导白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)分泌,并减弱葡萄糖转运蛋白4(GLUT-4)表达。GDM胎盘PPARα减少,并通过核因子κB(NF-κB)核转运过程抑制炎症反应。4-苯基丁酸(4-PBA)降低CHOP并增加PPARα,且降低我们类GDM外植体中的IL-6和TNF-α。然而,4-PBA和MK886处理后,我们发现CHOP表达无显著差异。在HTR8/Svneo细胞中,TG处理使PPARα水平降低,细胞核中NF-κB p65水平升高。敲低CHOP增加PPARα并降低NF-κB p65,而用TG处理HTR8细胞时,应用CHOP后PPARα表达下降,NF-κB p65表达升高。
ERS通过CHOP-PPARα-NF-κB信号通路诱导炎症异常激活和胰岛素抵抗,从而导致妊娠期GDM的病理生理改变。