Shen Xue-Yan, Huang Jing, Chen Li-Li, Sha Man-Ting, Gao Jing, Xin Hong
Department of Obstetrics, The Fourth Hospital of Shijiazhuang, Hebei Medical University, 16 Tangu North Street, Shijiazhuang City, Hebei Province, P.R. China.
Department of Obstetrics, The Second Hospital of Hebei Medical University, No.215, Heping West Road, Shijiazhuang, 050000, Hebei Province, P.R. China.
J Assist Reprod Genet. 2024 Nov;41(11):3201-3212. doi: 10.1007/s10815-024-03256-w. Epub 2024 Sep 17.
Abnormal cell death due to superficial trophoblast dysfunction caused by placental hypoxia plays a vital role in the development of preeclampsia (PE). Lactic acid stimulates gene transcription in chromatin through lactate modification of histone lysine. Nevertheless, the content and function of lactate in PE development remains largely unclear.
The contents of lactic acid and copper in 30 PE and 30 normal placentas were determined by kit colorimetry. Real-time quantitative fluorescent PCR (qRT-PCR) and Western blot were used to detect the expression of SLC31A1 in cells and tissues. Cell proliferation, apoptosis, and invasion were detected by cell counting kit 8 (CCK-8), MTS assay, colony formation assay, and Transwell assay. The transcriptional regulation between Grhl2 and SLC31A was verified by the luciferase reporter gene method and ChIP. The H3K18la modification level was detected by ChIP-PCR.
Herein, we detected increased lactic acid levels in the PE placental tissue, which inhibit the proliferation and invasion of trophoblasts. Interestingly, lactic acid increases intracellular copper content by enhancing the expression of SLC31A1, a key protein of copper ion transporters. Lentivirus knockdown of SLC31A1 blocked the lactate-induced proliferation and invasion of trophoblasts by inhibiting cell cuproptosis. Mechanically, we identified that Grhl2 mediated SLC31A1 expression through transcription and participated in SLC31A1-inhibited proliferation, invasion, and cuproptosis of trophoblasts. Furthermore, the high lactate content increased Grhl2 expression by enhancing lactate modification of histone H3K18 in the Grhl2 promoter region.
Blocking the lactate-regulated Grhl2/SLC31A1 axis and trophoblastic cuproptosis may be a potential approach to prevent and treat PE.
胎盘缺氧导致的浅表滋养层功能障碍引起的异常细胞死亡在子痫前期（PE）的发生发展中起重要作用。乳酸通过对组蛋白赖氨酸进行乳酸修饰来刺激染色质中的基因转录。然而，乳酸在PE发生发展中的含量及功能仍不清楚。
采用试剂盒比色法测定30例PE胎盘组织和30例正常胎盘组织中乳酸和铜的含量。采用实时定量荧光PCR（qRT-PCR）和蛋白质免疫印迹法检测细胞和组织中SLC31A1的表达。采用细胞计数试剂盒8（CCK-8）、MTS法、集落形成试验和Transwell试验检测细胞增殖、凋亡和侵袭情况。采用荧光素酶报告基因法和染色质免疫沉淀法（ChIP）验证Grhl2与SLC31A之间的转录调控。采用ChIP-PCR检测H3K18la修饰水平。
在此，我们检测到PE胎盘组织中乳酸水平升高，其抑制滋养层细胞的增殖和侵袭。有趣的是，乳酸通过增强铜离子转运关键蛋白SLC31A1的表达来增加细胞内铜含量。慢病毒敲低SLC31A1可通过抑制细胞铜死亡来阻断乳酸诱导的滋养层细胞增殖和侵袭。机制上，我们发现Grhl2通过转录介导SLC31A1表达，并参与SLC31A1抑制的滋养层细胞增殖、侵袭和铜死亡。此外，高乳酸含量通过增强Grhl2启动子区域组蛋白H3K18的乳酸修饰来增加Grhl2表达。
阻断乳酸调节的Grhl2/SLC31A1轴和滋养层细胞铜死亡可能是预防和治疗PE的潜在途径。
