Chu Xiaodan, Xu Jie, Shen Xinggui, Sheng Wenji, Sun Jingxia, Gu Yang, Lewis David F, Cooper Danielle, Zoorob Dani, Wang Yuping
Department of Obstetrics and Gynecology, Louisiana State University Health Sciences Center, Shreveport, LA 71103, USA.
Department of Obstetrics and Gynecology, The Second Affiliated Hospital, Harbin Medical University, Harbin 150086, China.
Pathophysiology. 2025 Mar 4;32(1):10. doi: 10.3390/pathophysiology32010010.
BACKGROUND/OBJECTIVES: Hydrogen sulfide (HS) is a vasorelaxant gas and exerts anti-oxidative, anti-inflammatory, and cytoprotective effects. HS has been implicated in regulating placental vaso-activity and angiogenesis. It is believed that abnormal trophoblast production of vasodilators and angiogenic factors contributes to pre-eclampsia development. However, little is known about whether aberrant HS production is present in placental trophoblasts from pre-eclamptic pregnancies.
Trophoblasts were isolated from normal and pre-eclamptic placentas. After incubation, cell production of HS in the culture medium and the cellular levels of HS were analyzed by reversed phase high-performance liquid chromatography (RP-HPLC). Expression levels of the three key HS converting enzymes, cystathionine-β-synthase (CBS), cystathionine-γ-lyase (CSE), and 3-mercaptopyruvate sulfurtransferase (3-MST), were determined by immunohistochemistry. The protein expression of CBS and CSE was assessed by Western blot analysis.
(1) Trophoblast production and cellular levels of HS were significantly reduced in cells from pre-eclamptic vs. normal placentas; (2) free HS production was increased in a time-dependent manner in cultured trophoblasts from normal, but not from pre-eclamptic, placentas; and (3) strong CBS and CSE expression was seen in trophoblasts from normal, as opposed to pre-eclamptic, placentas. Reduced CBS and CSE expression in trophoblasts from pre-eclamptic vs. normal placentas were confirmed by Western blot analysis; and (4) 3-MST expression was undetachable in both normal and pre-eclamptic placentas, but 3-MST expression was strongly expressed in the first and second trimester placentas.
These data provide plausible evidence that downregulation of CBS and CSE, but not 3-MST, expression may be responsible for reduced free HS production and decreased cellular HS levels in pre-eclamptic placentas. Our data provide further evidence that expression of 3-MST in placental trophoblasts is likely gestational age (developmental)-dependent.
背景/目的：硫化氢（HS）是一种血管舒张性气体，具有抗氧化、抗炎和细胞保护作用。HS与调节胎盘血管活性和血管生成有关。据信，滋养层细胞产生血管舒张剂和血管生成因子异常会导致子痫前期的发展。然而，对于子痫前期妊娠胎盘滋养层细胞中HS产生异常的情况知之甚少。
从正常和子痫前期胎盘中分离出滋养层细胞。培养后，通过反相高效液相色谱法（RP-HPLC）分析培养基中HS的细胞产生量以及细胞内HS水平。通过免疫组织化学法测定三种关键HS转化酶，即胱硫醚-β-合酶（CBS）、胱硫醚-γ-裂解酶（CSE）和3-巯基丙酮酸硫转移酶（3-MST）的表达水平。通过蛋白质印迹分析评估CBS和CSE的蛋白质表达。
（1）与正常胎盘来源的细胞相比，子痫前期胎盘来源的细胞中HS的细胞产生量和细胞内水平显著降低；（2）正常胎盘来源的培养滋养层细胞中，游离HS的产生量随时间呈依赖性增加，而子痫前期胎盘来源的培养滋养层细胞则不然；（3）正常胎盘来源的滋养层细胞中可见强烈的CBS和CSE表达，与之相反，子痫前期胎盘来源的滋养层细胞中则不然。蛋白质印迹分析证实，子痫前期胎盘来源的滋养层细胞中CBS和CSE表达降低；（4）在正常和子痫前期胎盘中均未检测到3-MST表达，但在妊娠早期和中期胎盘中3-MST表达强烈。
这些数据提供了合理的证据，表明CBS和CSE而非3-MST表达下调可能是子痫前期胎盘中游离HS产生减少和细胞内HS水平降低的原因。我们的数据进一步证明，胎盘滋养层细胞中3-MST的表达可能与胎龄（发育）有关。
