Department of Obstetrics and Gynecology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Republic of Korea.
Sungkyunkwan University School of Medicine, Samsung Biomedical Research Institute, Samsung Medical Center, Seoul, Republic of Korea.
Sci Rep. 2020 Aug 20;10(1):14033. doi: 10.1038/s41598-020-70959-2.
Preeclampsia is one of the most serious complications of pregnancy, affecting 5-10% of parturients worldwide. Recent studies have suggested that autophagy is involved in trophoblast invasion and may be associated with defective placentation underlying preeclampsia. We thus aimed to understand the mechanistic link between autophagy and trophoblast invasion. Using the two most commonly used trophoblast cell lines, JEG-3 and HTR-8/SVneo, we inhibited autophagy by ATG5 and beclin-1 shRNA. Conversion of LC3-II was evaluated in ATG5 and beclin-1 knock-down cells in the presence of the lysosomal protease inhibitors E-64d and pepstatin A, to detect the efficiency of autophagy inhibition. Upon autophagy inhibition, we measured cell invasion, activity of NF-κB and related signaling pathways, MMP-2, MMP-9, sFlt-1, and TNF-α levels. Autophagy inhibition increased the invasiveness of these trophoblastic cell lines and increased Akt and NF-κB activity as well as p65 expression. Of note, an NF-κB inhibitor significantly attenuated the trophoblast invasion induced by autophagy inhibition. Autophagy inhibition was also associated with increased MMP-2 and MMP-9 levels and decreased the production of sFlt-1 and TNF-α. Collectively, our results indicate that autophagy regulates trophoblast invasiveness in which the NF-κB pathway and MMP-2, MMP-9, sFlt-1 and TNF-α levels are affected.
子痫前期是妊娠最严重的并发症之一，影响全球 5-10%的产妇。最近的研究表明自噬参与滋养细胞浸润，可能与子痫前期基础的胎盘形成缺陷有关。因此，我们旨在了解自噬与滋养细胞浸润之间的机制联系。我们使用两种最常用的滋养层细胞系 JEG-3 和 HTR-8/SVneo，通过 ATG5 和 beclin-1 shRNA 抑制自噬。在溶酶体蛋白酶抑制剂 E-64d 和胃蛋白酶抑制剂 A 的存在下，检测 LC3-II 的转化，以检测自噬抑制的效率。在自噬抑制后，我们测量了细胞侵袭、NF-κB 及其相关信号通路的活性、MMP-2、MMP-9、sFlt-1 和 TNF-α 水平。自噬抑制增加了这些滋养层细胞系的侵袭性，并增加了 Akt 和 NF-κB 活性以及 p65 表达。值得注意的是，NF-κB 抑制剂显著减弱了自噬抑制诱导的滋养细胞侵袭。自噬抑制还与 MMP-2 和 MMP-9 水平升高以及 sFlt-1 和 TNF-α 产生减少有关。总之，我们的结果表明自噬调节滋养细胞的侵袭性，其中 NF-κB 途径和 MMP-2、MMP-9、sFlt-1 和 TNF-α 水平受到影响。
