沉默GATA4对多囊卵巢综合征大鼠模型卵巢颗粒细胞的影响  

Effects of silencing GATA4 on ovarian granulosa cells in rats with polycystic ovary syndrome

作　　者：贺聪 黄亚妮 刘晨晨 袁冬[3] 邵金坤 寇灵宙 HE Cong;HUANG Yani;LIU Chenchen;YUAN Dong;SHAO Jinkun;KOU Lingzhou(Department of Oncology,Zhongxiang People's Hospital/Zhongxiang Hospital of People's Hospital Affiliated to Wuhan University,Hubei Jingmen 431900,China.;Cardiac Function Room,Zhongxiang People's Hospital/Zhongxiang Hospital of People's Hospital Affiliated to Wuhan University,Hubei Jingmen 431900,China.;Department of Radiology Imaging,Zhongxiang People's Hospital/Zhongxiang Hospital of People's Hospital Affiliated to Wuhan University,Hubei Jingmen 431900,China.;Department of Ultrasound Imaging,Zhongxiang People's Hospital/Zhongxiang Hospital of People's Hospital Affiliated to Wuhan University,Hubei Jingmen 431900,China.)

机构地区：[1]钟祥市人民医院/武汉大学人民医院钟祥医院肿瘤科,湖北荆门431900 [2]钟祥市人民医院/武汉大学人民医院钟祥医院心功能室,湖北荆门431900 [3]钟祥市人民医院/武汉大学人民医院钟祥医院放射影像科,湖北荆门431900 [4]钟祥市人民医院/武汉大学人民医院钟祥医院超声影像科,湖北荆门431900

出　　处：《现代肿瘤医学》2024年第19期3665-3670,共6页Journal of Modern Oncology

摘　　要：目的:探讨沉默GATA结合蛋白4(GATA-binding protein 4,GATA4)表达对多囊卵巢综合征(polycystic ovary syndrome,PCOS)大鼠模型卵巢颗粒细胞增殖与凋亡、分泌功能及炎症反应的影响及相关机制。方法:制备PCOS模型,分离培养PCOS模型大鼠卵巢颗粒细胞并将其分为对照组(Ctrl组)、siRNA-NC组、siRNA-GATA4组,检测转染后三组卵巢颗粒细胞细胞中GATA4表达和细胞增殖与凋亡情况,采用酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测细胞上清液中激素和炎症因子含量,Western Blotting法检测细胞中活化的半胱氨酸天冬氨酸蛋白酶3(cleaved caspase-3)、磷脂酰肌醇-3-激酶(phosphatidylinositol-3-kinase,PI3K)、磷酸化的丝氨酸苏氨酸激酶(p-serine threonine kinase,p-AKT)蛋白表达。结果:siRNA-GATA4组卵巢颗粒细胞增殖率、GATA4、PI3K、p-AKT蛋白表达量较Ctrl组明显减少,细胞凋亡率、cleaved caspase-3蛋白表达量、细胞上清液中雌二醇(estradiol,E2)、黄体生成素(luteinizing hormone,LH)、睾酮(testosterone,T)、白细胞介素6(interleukin-6,IL-6)、IL-1β及肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)水平较Ctrl组明显升高,差异有统计学意义(P<0.05),siRNA-NC组与Ctrl组卵巢颗粒细胞增殖率、细胞凋亡率、GATA4、cleaved caspase-3、E2、LH、T、IL-6、IL-1β、TNF-α、PI3K及p-AKT比较,差异无统计学意义(P>0.05)。结论:沉默PCOS大鼠模型中GATA4表达,可抑制卵巢颗粒细胞增殖,促进其凋亡,加剧激素分泌异常和炎症反应,其机制可能与下调PI3K/AKT信号通路有关。Objective:To explore the effects and related mechanisms of silencing GATA-binding protein 4(GATA4)on proliferation and apoptosis of ovarian granulosa cells,secretory function and inflammatory response in rats with polycystic ovary syndrome(PCOS).Methods:PCOS models were prepared.The ovarian granulosa cells in PCOS rats were cultured and divided into control group(Ctrl group),siRNA-NC group and siRNA-GATA4 group.The expression of GATA4 in ovarian granulosa cells,cells proliferation and apoptosis in the three groups were detected after transfection.The levels of hormones and inflammatory factors in cell supernatant were detected by enzyme-linked immunosorbent assay(ELISA).The expressions of activated cysteine aspartate protease 3(cleaved caspase-3),phosphatidylinositol-3-kinase(PI3K)and phosphorylated-serine threonine kinase(p-AKT)proteins were detected by Western Blotting.Results:The proliferation rate of ovarian granuleous cells,expressions of GATA4,PI3K and P-Akt in siRNA-GATA4 group were significantly lower than those in Ctrl group,while apoptosis rate,expression of cleaved caspase-3 and levels of estradiol(E2),luteinizing hormone(LH),testosterone(T),interleukin-6(IL-6),IL-1βand tumor necrosis factor-α(TNF-α)in cell supernatant were significantly higher than those in Ctrl group(P<0.05).There was no significant difference in proliferation rate of ovarian granuleous cells,apoptosis rate,expressions of GATA4,cleaved caspase-3,PI3K and P-Akt proteins or levels of E2,LH,T,IL-6,IL-1βand TNF-αbetween siRNA-NC group and Ctrl group(P>0.05).Conclusion:Silencing GATA4 can inhibit proliferation and promote apoptosis of ovarian granulosa cells,aggravate abnormal hormones secretion and inflammatory response in PCOS rats,and the mechanisms may be related to down-regulating PI3K/AKT signaling pathways.

关 键 词：多囊卵巢综合征 大鼠模型 卵巢颗粒细胞 GATA结合蛋白4 增殖 凋亡 半胱氨酸天冬氨酸蛋白酶3 磷脂酰肌醇-3-激酶/丝氨酸苏氨酸激酶信号通路 

分 类 号：R733.31[医药卫生—肿瘤]