目的 明确尿酸和常见高尿酸治疗药物对人绒毛膜癌（Bewo）细胞活力及融合功能的影响。方法 在加入或不加入50 μmol·L^-1毛喉素诱导Bewo细胞融合的情况下，分别使用（30、50、70、100、160、200、400、600 mg·L^-1）尿酸或者别嘌呤醇、非布司他、苯溴马隆、丙磺舒（1、10、20、50、100 μmol·L^-1）处理Bewo细胞48 h，通过CCK-8法检测细胞活力并筛选药物合适浓度，采用酶联免疫吸附测定（ELISA）法检测人绒毛膜促性腺激素（hCG）分泌量；使用激光共聚焦显微镜观察细胞融合情况并计算融合率；通过实时荧光定量PCR（qRT-PCR）技术分析融合关键基因合胞素1（ERVW-1）、合胞素2（ERVFRD-1）、合胞素2受体（MFSD2A）、丙氨酸/丝氨酸/半胱氨酸/苏氨酸转运蛋白（ASCT1）、ASCT2 mRNA表达量。结果 各浓度尿酸对Bewo细胞活力没有明显影响；与对照组相比，100 μmol·L^-1别嘌呤醇组的Bewo细胞活力显著降低（P＜0.05）；在加入毛喉素后，100 μmol·L^-1别嘌呤醇组的Bewo细胞活力在一定程度上下降，但没有显著差异；加或不加毛喉素，100 μmol·L^-1非布司他，50、100 μmol·L^-1苯溴马隆都显著降低Bewo细胞活力（P＜0.05、0.001）。与毛喉素组相比，尿酸及4种高尿酸治疗药物均显著降低hCG分泌量（P＜0.01、0.001），显著抑制细胞的融合（P＜0.001）；尿酸显著抑制ERVW-1、ERVFRD-1、MFSD2A mRNA表达（P＜0.05、0.01、0.001），4种高尿酸治疗药物显著抑制VFRD-1、MFSD2A mRNA表达（P＜0.001），非布司他显著抑制ERVW-1 mRNA表达（P＜0.05），别嘌呤醇、非布司他、丙磺舒显著上调ASCT2的mRNA表达（P＜0.01、0.001）。结论 尿酸降低滋养细胞融合程度进而导致其分泌hCG功能下降；高尿酸药物会加剧这种功能的缺陷，针对于高尿酸孕妇急需开发新药。

Objective To clarify the effects of uric acid and common hyperuricemia treatment drugs on the viability of human choriocarcinoma (Bewo) cells and their fusion-related functions. Methods Bewo cells were treated with (30, 50, 70, 100, 160, 200, 400, 600 mg·L^-1) uric acid or allopurinol, febuxostat, benzbromarone, probenecid (1, 10, 20, 50, 100 μmol·L^-1) for 48 h with or without 50 μmol·L^-1 thapsigargin-induced fusion. Cell viability was detected by CCK-8 assay and the appropriate drug concentrations were screened. The secretion of human chorionic gonadotropin (hCG) was detected by ELISA. Cell fusion was observed by laser confocal microscopy and the fusion rate was calculated. The mRNA expression of key fusion genes, syncytin-1 (ERVW-1), syncytin-2 (ERVFRD-1), syncytin-2 receptor (MFSD2A), alanine/serine/cysteine/threonine transporter (ASCT1), and ASCT2 was analyzed by realtime fluorescence quantitative PCR (qRT-PCR). Results Uric acid at various concentrations had no significant effect on the viability of Bewo cells. Compared with the control group, the viability of Bewo cells in the 100 μmol·L^-1 allopurinol group was significantly reduced (P < 0.05). After the addition of thapsigargin, the viability of Bewo cells in the 100 μmol·L^-1 allopurinol group decreased to some extent, but there was no significant difference. With or without thapsigargin, 100 μmol·L^-1 febuxostat, 50 and 100 μmol·L^-1 benzbromarone significantly reduced the viability of Bewo cells (P < 0.05, 0.001). Compared with the thapsigargin group, uric acid and four hyperuricemia treatment drugs significantly reduced the secretion of hCG (P < 0.01, 0.001) and significantly inhibited cell fusion (P < 0.001). Uric acid significantly inhibited the mRNA expression of ERVW-1, ERVFRD-1, and MFSD2A (P < 0.05, 0.01, 0.001). The four hyperuricemia treatment drugs significantly inhibited the mRNA expression of ERVFRD-1 and MFSD2A (P < 0.001). Febuxostat significantly inhibited the mRNA expression of ERVW-1 (P < 0.05). Allopurinol, febuxostat, and probenecid significantly upregulated the mRNA expression of ASCT2 (P < 0.01, 0.001). Conclusion Uric acid can decrease the degree of fusion of trophoblast cell line and lead to the decrease of hCG secretory function. However, hyperuricemic drugs may exacerbate this functional deficit, and there is an urgent need to develop new drugs for pregnant women with high uric acid.

R965

青海省科学技术厅自然科学基金资助项目（2024-ZJ-923）；青海省“昆仑英才-高端创新创业人才”项目