摘要
目的急性青光眼期间视网膜神经节细胞(retinal ganglion cells,RGCs)的死亡会导致视网膜神经的进行性变性和不可逆的失明。葛根素被证实具有抗氧化和抗神经炎症的特性,可保护视网膜免受急性青光眼的伤害。此研究旨在探索其作用机制。方法构建急性青光眼大鼠模型,将大鼠随机分为5组:正常对照组(normal)、SnPP组、I/R组、葛根素(100μmol/L)组和葛根素+SnPP组。I/R、葛根素和葛根素+SnPP组的大鼠采用苏木精和伊红染色评价视网膜状况。ELISA和qRT-PCR法检测TNF-α、IL-1β的水平,蛋白质印迹法检测cleaved caspase-8、Nrf2、HO-1蛋白水平。免疫荧光检测Brn-3a、β3-tubulin和RBPMS的表达;流式细胞术分别检测细胞凋亡和细胞内活性氧的水平。结果葛根素治疗后显著减轻了急性青光眼大鼠的视网膜损伤和RGCs死亡,视网膜中活性氧和炎性细胞因子的产生显著减少。此外,葛根素处理直接降低了ROS的产生,并重新平衡了细胞内的氧化还原状态,从而有助于RGCs在体外的存活。葛根素治疗也减少了体外炎性细胞因子的产生。从机制上讲,葛根素通过调节Nrf2/HO-1通路,抑制RGCs的细胞凋亡和炎性细胞因子产生。此外,当使用HO-1抑制剂SnPP时,葛根素介导的急性青光眼的神经保护作用被消除。结论研究确定了RGCs凋亡的潜在机制,葛根素作为一种新的治疗剂,对急性青光眼具有神经保护作用。
Objective Death of retinal ganglion cells(RGCs)during acute glaucoma leads to progressive degeneration of retinal nerves and irreversible blindness.Puerarin has been shown to have antioxidant and anti-neuroinflammatory properties that protect the retina from acute glaucoma.However,the mechanism behind this process remains unclear.Methods In this study,rat model of acute glaucoma was established,and rats were randomly divided into five groups:normal control group,SnPP group,I/R group,puerarin(100μmol/L)group and puerarin+SnPP group.Hematoxylin and eosin staining were used to evaluate the retinal conditions in the I/R group,puerarin group and puerarin+SnPP group.Levels of TNF-αand IL-1βwere detected by ELISA and qRT-PCR.Expression levels of cleaved caspase-8,Nrf2,and HO-1 were detected by Western blotting.The expression levels of Brn-3a,β3-tubulin and RBPMS were detected by immunofluorescence.Cell apoptosis and intracellular reactive oxygen species were detected by flow cytometry.Results Puerarin treatment significantly reduced retinal damage and RGC death in acute glaucoma rats,and significantly reduced the production of reactive oxygen species and inflammatory cytokines in the retina.In addition,the reduction of ROS production by puerarin had re-balanced the intracellular redox status,thus contributing to the survival of RGCs in vitro.Puerarin treatment also reduced the production of inflammatory cytokines in vitro.Puerarin inhibited the cell apoptosis and the production of inflammatory cytokine in RGCs through regulation of Nrf2/HO-1 pathway.Moreover,the neuroprotective effect of puerarin on acute glaucoma was eliminated when SnPP,an HO-1 inhibitor,was administered.Conclusion Our findings identify the underlying mechanism of RGC apoptosis and the neuroprotective effect of a novel therapeutic agent puerarin on acute glaucoma.
作者
刘鹏
曹建
谢梅芬
张映平
彭菲
LIU Peng;CAO Jian;XIE Meifen;ZHANG Yingping;PENG Fei(Department of Ophthalmology,Xiangtan Central Hospital,Xiangtan,Hunan,411100,China;Physical Examination Center,Xiangtan Central Hospital,Xiangtan,Hunan,411100,China)
出处
《医学分子生物学杂志》
CAS
2023年第5期417-425,共9页
Journal of Medical Molecular Biology
基金
湖南省医学科学科技攻关计划项目(No.2018FFC236)。
