水飞蓟宾通过PI3K/Akt/mTOR信号通路降低蓝光诱导的视网膜神经节细胞凋亡和炎症研究  

Silybin reduces blue-induced apoptosis and inflammation of retinal ganglion cells through PI3K/Akt/mTOR signaling pathway

作　　者：包娅利 张惠君 孔宁 Bao Yali;Zhang Huijun;Kong Ning(Panyu Central Hospital,Guangzhou Medical University,Guangzhou,Guangdong 511400,China)

机构地区：[1]广州医科大学附属番禺中心医院,广东广州511400

出　　处：《首都食品与医药》2024年第24期11-15,共5页Capital Food Medicine

基　　金：广州市科技计划项目(编号:202201011689);番禺区科技计划项目(编号:2020-Z04-039)。

摘　　要：目的探究水飞蓟宾对蓝光诱导的视网膜神经节细胞(RGC)损伤所展现出的防护效应.方法采用蓝光照射制备RGC体外损伤模型,将其随机分成处理组(蓝光照射+RGC)和实验组(蓝光照射+RGC+水飞蓟宾),另外设立了一组对照组,为正常培养条件下生长的、未经任何特定干预措施的RGC.通过MTT法来量化细胞活力;利用流式细胞术和TUNEL染色分析测定细胞凋亡的比例和凋亡细胞的形态特征;运用ELISA法检测关键炎症因子TNF-α、IL-1β、IL-6以及抗炎因子IL-10的表达水平;借助Western blot技术分析与细胞凋亡密切相关的蛋白表达量以及PI3K/Akt/mTOR信号通路中关键蛋白的表达情况.结果蓝光照射导致RGC活力降低,而水飞蓟宾(50、100μM)处理可以显著降低蓝光照射引起的RGC损伤.Western blot、流式细胞术和TUNEL染色结果统一显示,水飞蓟宾可以大部分挽救蓝光照射引起的RGC凋亡.在蓝光照射过程中,RGC中TNF-α、IL-1β和IL-6的分泌量上调,IL-10的分泌量下调;而水飞蓟宾处理展现出对这些炎症因子表达的显著调控作用.蓝光照射会抑制RGC中PI3K/Akt/mTOR的表达,而水飞蓟宾处理则可以再次激活PI3K/Akt/mTOR信号通路.结论水飞蓟宾能够显著减轻蓝光诱导的RGC凋亡进程及伴随的炎症反应,其潜在的保护机制可能归因于对PI3K/Akt/mTOR信号通路的激活.Objective To investigate the protective effect of silybin on blue-induced retinal ganglion cell(RGC)damage.Methods The damage model of RGC in vitro was prepared by blue light irradiation and randomly divided into treatment group(blue light irradiation+RGC)and experimental group(blue light irradiation+RGC+silybin).In addition,a control group was set up,which was RGC growing under normal culture conditions without any specific intervention measures.Cell viability was quantified by MTT assay.Flow cytometry and TUNEL staining were used to determine the proportion of apoptosis and the morphological characteristics of apoptotic cells.The expression levels of key inflammatory factors TNF-α,IL-1β,IL-6 and anti-inflammatory factor IL-10 were detected by ELISA.Western blot technique was used to analyze the expression levels of proteins closely related to apoptosis and the expression of key proteins in the PI3K/Akt/mTOR signaling pathway.Results Blue light irradiation reduced RGC viability,and Silibin(50,100μM)treatment significantly reduced RGC damage caused by blue light irradiation.The results of Western blot,flow cytometry and TUNEL staining showed that silybin could mostly save the apoptosis of RGC induced by blue light irradiation.During blue light irradiation,the secretion of TNF-α,IL-1βand IL-6 in RGC was up-regulated,and the secretion of IL-10 was down-regulated.Silybin treatment showed significant regulatory effects on the expression of these inflammatory factors.Blue light irradiation can inhibit the expression of PI3K/Akt/mTOR in RGC,while silybin treatment can reactivate the PI3K/Akt/mTOR signaling pathway.Conclusion Silybin can significantly reduce the apoptosis process of RGC induced by blue light and the associated inflammatory response,and its potential protective mechanism may be attributed to the activation of PI3K/Akt/mTOR signaling pathway.

关 键 词：水飞蓟宾 青光眼 视网膜神经节细胞 细胞凋亡 炎症反应 

分 类 号：R775[医药卫生—眼科]