补肾活血方对aCL诱导滋养细胞损伤的保护作用及对P2X7介导的NLRP3通路的影响  

Protective effect of Bushen Huoxuefang on aCL-induced damage of trophoblasts and impact on P2X7-mediated NLRP3 pathways

作　　者：宋艳丽[1] 崔天薇 任杏杏[1] 石少琦 万彦荣[1] 张超男 王会娟 卫爱武[1] SONG Yan-li;CUI Tian-wei;REN Xing-xing;SHI Shao-qi;WAN Yan-rong;ZHANG Chao-nan;WANG Hui-juan;WEI Ai-wu(The First Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou,Henan 450002,China;不详)

机构地区：[1]河南中医药大学第一附属医院生殖医学科,河南郑州450002 [2]河南中医药大学第一临床医学院,河南郑州450002

出　　处：《中华医院感染学杂志》2025年第1期7-12,共6页Chinese Journal of Nosocomiology

基　　金：国家自然科学基金(82104911);河南中医药大学第一附属医院博士科研基金(KY-B0367-20);中国博士后科学基金第四批特别资助(站前)(2022TQ0099);中国博士后科学基金第72批面上资助二等(2022M721064)。

摘　　要：目的探索补肾活血方-丹寿汤对抗心磷脂抗体(aCL)诱导滋养细胞损伤的保护作用及对嘌呤能离子通道型7(P2X7)介导的Nod样受体蛋白3(NLRP3)通路的影响。方法建立aCL诱导滋养细胞损伤模型,通过CCK-8检测各组细胞活力,计算滋养细胞模型的最佳aCL-Ig G剂量,分别给予高、中、低浓度丹寿汤含药血清治疗,探索丹寿汤含药血清合适的治疗剂量,选择高浓度丹寿汤含药血清为最佳干预剂量,分别给予丹寿汤含药血清、P2X7受体拮抗剂(BBG)处理aCL诱导的模型细胞,TUNEL染色法检测各组细胞凋亡情况,酶联免疫吸附试验检测细胞中白细胞介素(IL)-1β、IL-18含量,免疫印迹法(WB)检测cleavedcaspase-1、ASC、NLRP3、Bcl2、Bax、cleavedcaspase-3、P2X7蛋白的表达。结果与Control组比较,aCL组细胞凋亡数目,细胞中IL-1β(59.08±9.42)ng/ml、IL-18(621.65±103.51)ng/ml,cleavedcaspase-1、ASC、NLRP3、Bax、cleavedcaspase-3、P2X7蛋白表达明显增加(均P<0.05),Bcl2蛋白水平明显降低(P<0.05);与aCL组比较,aCL+BBG组细胞中IL-1β(20.63±2.84)ng/ml、IL-18(263.71±47.55)ng/ml,cleavedcaspase-1、ASC、NLRP3、Bax、cleavedcaspase-3、P2X7蛋白表达明显降低(均P<0.05),Bcl2蛋白水平明显增加(P<0.05);aCL+丹寿汤组细胞存活率提高(P<0.05),细胞凋亡数目,细胞中IL-1β(33.04±3.71)ng/ml、IL-18(352.03±66.23)ng/ml,cleavedcaspase-1、ASC、NLRP3、Bax、cleavedcaspase-3、P2X7蛋白表达明显降低(P<0.05),Bcl2蛋白水平明显增加(P<0.05)。结论丹寿汤能抑制血小板活化,下调P2X7的表达,抑制NLRP3的激活,抑制滋养细胞的损伤,可能为减少流产的发生提供理论依据。OBJECTIVE To explore the protective effect of Bushenhuoxue Fang-Danshou decoction on anti-cardiolipin(aCL)-induced damage of trophoblasts and observe the impact on purinergic ion channel type 7(P2X7)-mediated Nod-like receptor protein 3(NLRP3)pathways.METHODS The model of aCL-induced damage of trophocytes was established,the cell viability was determined by CCK-8,the optimal dose of aCL-IgG of the trophoblast model was calculated.The high,medium and low concentrations of Danshou decoction-medicated serum were respectively administrated to explore the appropriate therapeutic dose of Danshou decoction-medicated serum,the high concentration of Danshou decoction-medicated serum was chosen as the optimal intervention dose,Danshou decoction-mediated serum and purinergic ion channel type 7(P2X7)receptor antagonist(BBG)were respectively given to treat the aCL-induced model cells,the apoptosis of the cells was detected by TUNEL staining method.The levels of interleukin(IL)-1βand IL-18 in the cells were detected by enzyme-linked immunosorbent assay(ELISA);the expression levels of cleaved caspase-l,ASC,NLRP3,Bcl2,Bax,cleaved caspase-3 and P2X7 proteins were detected by Western blot(WB).RESULTS The number of apoptotic cells and expression levels of IL-1β(59.08±9.42)ng/ml,IL-18(621.65±103.51)ng/ml,cleaved caspase-1,ASC,NLRP3,Bax,cleaved caspase-3 and P2X7 proteins were remarkably higher in the aCL group than in the Control group(P<0.05),while the Bcl2 protein level of the aCL group was remarkably lower than that of the Control group(P<0.05).The expression levels of IL-1β(20.63±2.84)ng/ml,IL-18(263.71±47.55)ng/ml,cleaved caspase-1,ASC,NLRP3,Bax,cleaved caspase-3 and P2X7 of the aCL+BBG group were remarkably lower than those of the aCL group(P<0.05),while the Bcl2 level of the aCL+BBG group was remarkably higher than that of the aCL group(P<0.05).The survival rate of cells of the aCL+Danshou decoction group was higher than that of the aCL group(P<0.05);the number of apoptotic cells and expression levels of IL-1β(33.04±

关 键 词：补肾活血方 丹寿汤 抗心磷脂抗体 嘌呤能离子通道型7 Nod样受体蛋白3 

分 类 号：R-33[医药卫生]