小檗碱调节ROS/TXNIP/NLRP3通路对高糖诱导的视网膜神经节细胞凋亡的影响OA
Impact of berberine on high glucose induced apoptosis of retinal ganglion cells by regulating the ROS/TXNIP/NLRP3 pathway
目的 探讨小檗碱调节活性氧(ROS)/硫氧还蛋白结合蛋白(TXNIP)/NOD样受体相关蛋白3(NLRP3)通路对高糖诱导的视网膜神经节细胞(RGCs)凋亡的影响.方法 将RGC-5细胞分为对照组、高糖组、小檗碱组、空载质粒(pcDNA-NC)组、TXNIP过表达质粒(pcDNA-TXNIP)组及小檗碱+pcDNA-TXNIP组,细胞活力检测用细胞计数试剂盒(CCK-8)法,细胞凋亡率检测用流式细胞术,细胞裂解液中丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、超氧化物岐化酶(SOD)、白细胞介素1β(IL-1β)、白细胞介素18(IL-18)水平检测用酶联免疫吸附法(ELISA),ROS检测用2,7-二氯荧光素二乙酸酯(DCFH-DA)法,Bcl-2相关X蛋白(Bax)、Caspase-3、Caspase-9、TXNIP、NLRP3蛋白表达检测用Western blot法.结果 与对照组相比,高糖组RGC-5细胞450 nm处的光密度(OD450)值、GSH-Px、SOD水平降低,细胞凋亡率、MDA、IL-1β、IL-18、ROS水平及Bax、Caspase-3、Caspase-9、TXNIP、NLRP3蛋白表达量升高,差异均有统计学意义(均P<0.05);与高糖组相比,小檗碱组RGC-5细胞OD450值、GSH-Px、SOD水平升高,细胞凋亡率、MDA、IL-1β、IL-18、ROS水平及凋亡和TXNIP/NLRP3通路蛋白表达量降低,差异均有统计学意义(均P<0.05);pcDNA-TXNIP组RGC-5细胞OD450值、GSH-Px、SOD水平下降,但细胞凋亡率、MDA、IL-1β、IL-18、ROS 水平及凋亡蛋白、TXNIP/NLRP3 通路蛋白表达量增加(均 P<0.05);与小檗碱组相比,小檗碱+pcDNA-TXNIP组RGC-5细胞OD450值、GSH-Px、SOD水平降低,而细胞凋亡率、MDA、IL-1β、IL-18、ROS水平及凋亡蛋白、TXNIP/NLRP3通路蛋白表达量升高(均P<0.05).结论 小檗碱可通过抑制ROS/TXNIP/NLRP3通路抑制高糖诱导RGCs凋亡.
Objective To investigate the impact of berberine on high glucose induced apoptosis of retinal gan-glion cells(RGCs)by regulating reactive oxygen species(ROS)/thioredoxin-binding protein(TXNIP)/NOD-like receptor pyrin domain containing 3(NLRP3)pathway.Methods RGC-5 cells were assigned into the control group,high glucose group,berberine group,negative control plasmid(pcDNA-NC)group,TXNIP overexpression plasmid(pcDNA-TXNIP)group,and berberine+pcDNA-TXNIP group.Cell viability was detected using the Cell Counting Kit(CCK-8)method.Cell apoptosis rate was detected using flow cytometry.The levels of malondialdehyde(MDA),glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),interleukin-1β(IL-1β),and interleukin-18(IL-18)in cell lysate were detected us-ing enzyme-linked immunosorbent assay(ELISA).The detection of ROS was performed using the 2',7'-Dichlorodihydro-fluorescein diacetate(DCFH-DA)method.Western blot was carried out to measure the expression of Bcl-2 associated X protein(Bax),Caspase-3,Caspase-9,TXNIP,and NLRP3 proteins.Results The high glucose group showed a decrease in optical density at 450 nm(OD450)value,GSH-Px,and SOD levels in RGC-5 cells,and an increase in apoptosis rate,MDA,IL-1β,IL-18,ROS levels,and Bax,Caspase-3,Caspase-9,TXNIP,and NLRP3 protein expression levels than the control group,and all differences were statistically significant(all P<0.05).The berberine group showed an increase in OD450 value,GSH-Px,and SOD levels in RGC-5 cells,and a decrease in apoptosis rate,MDA,IL-1β,IL-18,ROS levels and the apoptosis and TXNIP/NLRP3 pathway protein expression compared with the high glucose group,and all differences were statisticallg significant(all P<0.05);while the pcDNA-TXNIP group showed a decrease in OD450 value,GSH-Px,and SOD levels in RGC-5 cells,and an increase in apoptosis rate,MDA,IL-1β,IL-18,ROS levels,and the apoptosis protein and TXNIP/NLRP3 pathway protein expression(all P<0.05).The berberine+pcDNA-TXNIP group had a decrease in OD450 value,GSH-Px,and SOD levels in RGC-5 cells,and an increase in apoptosis rate,MDA,IL-1β,IL-18,ROS levels,and the apoptosis protein and TXNIP/NLRP3 pathway protein expression compared with the berberine group(all P<0.05).Conclusions Berberine can inhibit high glucose induced apoptosis of RGCs by suppressing the ROS/TXNIP/NLRP3 pathway.
高毓妍;高惠英
050091 河北石家庄,石家庄工商职业学院050000 河北石家庄,石家庄爱尔眼科医院小儿眼科
活性氧硫氧还蛋白结合蛋白NOD样受体相关蛋白3小檗碱高糖视网膜神经节细胞
Reactive oxygen speciesThioredoxin-binding proteinNOD-like receptor pyrin domain containing 3BerberineHigh glucoseRetinal ganglion cells
《临床眼科杂志》 2026 (2)
110-115,6
河北省医学科学研究重点课题计划项目(20230227)
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