目的 明確尿酸和常見高尿酸治療藥物對人絨毛膜癌（Bewo）細(xì)胞活力及融合功能的影響。方法 在加入或不加入50 μmol·L^-1毛喉素誘導(dǎo)Bewo細(xì)胞融合的情況下，分別使用（30、50、70、100、160、200、400、600 mg·L^-1）尿酸或者別嘌呤醇、非布司他、苯溴馬隆、丙磺舒（1、10、20、50、100 μmol·L^-1）處理Bewo細(xì)胞48 h，通過CCK-8法檢測細(xì)胞活力并篩選藥物合適濃度，采用酶聯(lián)免疫吸附測定（ELISA）法檢測人絨毛膜促性腺激素（hCG）分泌量；使用激光共聚焦顯微鏡觀察細(xì)胞融合情況并計算融合率；通過實時熒光定量PCR（qRT-PCR）技術(shù)分析融合關(guān)鍵基因合胞素1（ERVW-1）、合胞素2（ERVFRD-1）、合胞素2受體（MFSD2A）、丙氨酸/絲氨酸/半胱氨酸/蘇氨酸轉(zhuǎn)運蛋白（ASCT1）、ASCT2 mRNA表達(dá)量。結(jié)果 各濃度尿酸對Bewo細(xì)胞活力沒有明顯影響；與對照組相比，100 μmol·L^-1別嘌呤醇組的Bewo細(xì)胞活力顯著降低（P＜0.05）；在加入毛喉素后，100 μmol·L^-1別嘌呤醇組的Bewo細(xì)胞活力在一定程度上下降，但沒有顯著差異；加或不加毛喉素，100 μmol·L^-1非布司他，50、100 μmol·L^-1苯溴馬隆都顯著降低Bewo細(xì)胞活力（P＜0.05、0.001）。與毛喉素組相比，尿酸及4種高尿酸治療藥物均顯著降低hCG分泌量（P＜0.01、0.001），顯著抑制細(xì)胞的融合（P＜0.001）；尿酸顯著抑制ERVW-1、ERVFRD-1、MFSD2A mRNA表達(dá)（P＜0.05、0.01、0.001），4種高尿酸治療藥物顯著抑制VFRD-1、MFSD2A mRNA表達(dá)（P＜0.001），非布司他顯著抑制ERVW-1 mRNA表達(dá)（P＜0.05），別嘌呤醇、非布司他、丙磺舒顯著上調(diào)ASCT2的mRNA表達(dá)（P＜0.01、0.001）。結(jié)論 尿酸降低滋養(yǎng)細(xì)胞融合程度進(jìn)而導(dǎo)致其分泌hCG功能下降；高尿酸藥物會加劇這種功能的缺陷，針對于高尿酸孕婦急需開發(fā)新藥。

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

青海省科學(xué)技術(shù)廳自然科學(xué)基金資助項目（2024-ZJ-923）；青海省“昆侖英才-高端創(chuàng)新創(chuàng)業(yè)人才”項目