摘要
目的本研究旨在探讨高糖环境下活性氧(ROS)介导的胰岛β细胞焦亡对妊娠糖尿病(GDM)发生的潜在机制。方法采用小鼠胰岛β细胞进行体外实验,设置不同的实验组别:对照组、高糖组(HG)、Epal(醛糖还原酶抑制剂)组、FPS-ZM1(晚期糖化终产物受体抑制剂)组和DPI(NADPH氧化酶抑制剂)组。通过CCK-8法检测细胞活力,DCFH-DA荧光探针法测定ROS产生,微量法和可见分光光度法定量分析抗氧化酶超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性,Western blot检测焦亡相关蛋白表达,葡萄糖刺激胰岛素分泌实验检测胰岛素分泌水平。结果与Control组相比,HG组胰岛β细胞形态差,活力下降,ROS产生增加,抗氧化酶(SOD和GSH-Px)活性降低,焦亡相关蛋白表达升高,胰岛素分泌减少(P<0.05)。与HG组相比,同样在高糖环境处理下的Epal组、FPS-ZM1组和DPI组细胞生长状态、细胞活力、抗氧化酶活性和胰岛素分泌水平均有所改善,且ROS产生和焦亡相关蛋白表达均降低(P<0.05)。其中FPS-ZM1组和DPI组的改善效果均优于Epal组处理效果(P<0.05)。结论高糖环境通过ROS介导的途径加剧胰岛β细胞的损伤,导致GDM的发生,其内在机制可能主要是通过激活NADPH氧化酶途径、糖基化终末产物形成途径,以及抑制抗氧化酶活性来实现的。抑制ROS产生或其上游信号通路能够减轻胰岛β细胞的焦亡,为GDM的临床治疗提供了新的策略和潜在靶点。
Objective This study aims to investigate the potential mechanisms by which reactive oxygen species(ROS)-mediated pyroptosis of pancreaticβ-cells contributes to the development of gestational diabetes mellitus(GDM)under high-glucose conditions.Methods Mouse pancreaticβ-cells were used for in vitro experiments and different experimental groups were set up:Control,high-glucose(HG),Epal(aldose reductase inhibitor),FPS-ZM1(advanced glycation end-product receptor inhibitor),and DPI(NADPH oxidase inhibitor).Cell viability was assessed using the CCK-8 assay,ROS production was measured via DCFH-DA fluorescence,and activities of antioxidant enzymes superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px)were quantatively analyzed using microassay and visible spectrophotometry.Pyroptosis-related protein expression was evaluated by Western blot,and insulin secretion was determined through glucose-stimulated insulin secretion assays.Results Compared with the Control group,the HG group exhibited impaired cell morphology,reduced cell viability,increased ROS production,decreased antioxidant enzymes(SOD and GSH-Px)activities,elevated pyroptosis-related protein expression,and diminished insulin secretion(P<0.05).In contrast,the Epal group,FPS-ZM1 group,and DPI group,under high-glucose conditions,showed improved cell morphology,enhanced cell viability,increased antioxidant enzyme activities,and higher insulin secretion level,along with reduced ROS production and pyroptosis-related protein expression(P<0.05).The improvement effect of FPS-ZM1 group and DPIgroup was better than that of Epal group(P<0.05).Conclusion High-glucose conditions exacerbate pancreaticβ-cell damage and leads to the occurrence of GDM through ROS-mediated pathways,the internal mechanism may mainly be through activating NADPH oxidase pathway,the formation pathway of advanced glycation end-products,and suppressing the activity of antioxidant enzyme activities.Inhibiting ROS production or its upstream signaling pathways can mitigateβ-cell pyroptosis,offering novel strategies and potential therapeutic targets for the clinical management of GDM.
作者
陈蓓
田婷
CHEN Bei;TIAN Ting(Department of Obstetrics,The Second Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang,Shaanxi 712000,China)
出处
《中国优生与遗传杂志》
2025年第10期2135-2140,共6页
Chinese Journal of Birth Health & Heredity
基金
陕西省卫生计生科研基金项目(2016D029)。
关键词
妊娠糖尿病
高糖环境
活性氧
胰岛β细胞焦亡
胰岛素分泌
gestational diabetes mellitus
high-glucose environment
reactive oxygen species
pancreaticβ-cell apoptosis
insulin secretion
