Tlaye Kenean Getaneh, Woldeamanuel Gashaw Garedew, Wong Kenneth Chi-Yin, Chen Lu, Zheng Ruqun, So Pui Kin, Wang Xueqin, Nguyen-Hoang Long, Zhong Mei, So Hon Cheong, Leung Bo Wah, Huang Yu, Wang Yao, Poon Liona C, Wang Chi Chiu
Department of Obstetrics and Gynaecology (K.G.T., G.G.W., L.C., R.Z., X.W., L.N.-H., B.W.L., Y.W., L.C.P., C.C.W.), The Chinese University of Hong Kong, Shatin, Hong Kong.
Department of Biomedical Sciences, Wolkite University, Ethiopia (G.G.W.).
Circ Res. 2025 Jun 20;137(1):69-82. doi: 10.1161/CIRCRESAHA.124.325699. Epub 2025 May 7.
It has become evident that some women develop preeclampsia despite aspirin. This study aimed to examine how such aspirin nonresponsiveness develops in high-risk preeclampsia pregnancies by exploring the role of genetic polymorphisms and aspirin metabolism.
The study involved pregnant women who developed preeclampsia despite low-dose aspirin and those who did not. First, we conducted a pharmacogenomic association study exploring the association of potential genetic variants with aspirin nonresponsiveness. Next, we analyzed the rate of enzymatic aspirin hydrolysis in maternal plasma. The extent of placental exposure to acetylsalicylic acid and its bioactive metabolites, that is, salicylic acid and gentisic acid, was determined by liquid chromatography-mass spectrometry. The expressions of placental AMEs (aspirin metabolizing enzymes), that is, GLYAT (glycine-N-acyltransferase), UGT1A6 (UDP glucuronosyltransferase family 1 member A6), CYP2E1 (cytochrome P450 family 2 subfamily E member 1), and NAT2 (N-acetyltransferase 2) were analyzed by quantitative reverse transcription polymerase chain reaction, immunohistochemistry staining, and enzyme-linked immunosorbent assay. Finally, the effects of AMEs were further studied on HTR-8/SVneo (an immortalized human trophoblast cell line derived from first-trimester placenta) and hPCTB (human primary cytotrophoblast) cells.
Our genetic study showed that single-nucleotide polymorphisms (SNPs) of genes involved in aspirin pharmacokinetics and pharmacodynamics were not associated with aspirin nonresponsiveness in preeclampsia. Rates of aspirin hydrolysis in maternal plasma and the concentrations of acetylsalicylic acid, salicylic acid, and gentisic acid in the placenta did not differ between aspirin-responsive and aspirin-nonresponsive women. Intriguingly, GLYAT was significantly upregulated in the aspirin-nonresponsive placenta and associated with aspirin nonresponsiveness. This overexpression of GLYAT was found to diminish the proangiogenic, anti-inflammatory, and antisenescence effects of salicylic acid in HTR-8/SVneo and hPCTB cells.
Our study revealed that maternal genetic factors and plasma aspirin hydrolysis are not among the decisive factors in determining the effectiveness of low-dose aspirin in preventing preeclampsia among high-risk women. Instead, placental GLYAT appears to play a key role by limiting the effect of salicylic acid in the placenta.
已明显看出，一些女性即便服用了阿司匹林仍会发生先兆子痫。本研究旨在通过探究基因多态性和阿司匹林代谢的作用，来研究在高危先兆子痫妊娠中这种阿司匹林无反应性是如何发生的。
该研究纳入了尽管服用小剂量阿司匹林仍发生先兆子痫的孕妇以及未发生先兆子痫的孕妇。首先，我们进行了一项药物基因组关联研究，探究潜在基因变异与阿司匹林无反应性之间的关联。接下来，我们分析了母体血浆中阿司匹林的酶促水解速率。通过液相色谱 - 质谱法测定胎盘对乙酰水杨酸及其生物活性代谢物（即水杨酸和龙胆酸）的暴露程度。通过定量逆转录聚合酶链反应、免疫组织化学染色和酶联免疫吸附测定法分析胎盘阿司匹林代谢酶（AMEs），即甘氨酸 - N - 酰基转移酶（GLYAT）、尿苷二磷酸葡萄糖醛酸基转移酶1家族A6成员（UGT1A6）、细胞色素P450家族2亚家族E成员1（CYP2E1）和N - 乙酰转移酶2（NAT2）的表达。最后，进一步研究AMEs对HTR - 8/SVneo（一种源自孕早期胎盘的永生化人滋养层细胞系）和人原代细胞滋养层（hPCTB）细胞的影响。
我们的基因研究表明，参与阿司匹林药代动力学和药效学的基因单核苷酸多态性（SNP）与先兆子痫中的阿司匹林无反应性无关。阿司匹林反应者和无反应者之间，母体血浆中阿司匹林的水解速率以及胎盘中乙酰水杨酸、水杨酸和龙胆酸的浓度没有差异。有趣的是，GLYAT在阿司匹林无反应性胎盘中显著上调，且与阿司匹林无反应性相关。发现GLYAT的这种过表达会减弱水杨酸在HTR - 8/SVneo和hPCTB细胞中的促血管生成、抗炎和抗衰老作用。
我们的研究表明，母体遗传因素和血浆阿司匹林水解并非决定小剂量阿司匹林预防高危女性先兆子痫有效性的决定性因素。相反，胎盘GLYAT似乎通过限制水杨酸在胎盘中的作用而发挥关键作用。
