基于ROS/TXNIP/NLRP3信号通路探究山茱萸新苷对糖尿病视网膜病变大鼠的治疗效果  

Exploring the therapeutic effect of cornuside on diabetic retinopathy in rats based on the ROS/TXNIP/NLRP3 signaling pathway

作　　者：陈梅 秦学维 姚贤凤 李宗智 CHEN Mei;QIN Xuewei;YAO Xianfeng;LI Zongzhi(Department of Ophthalmology,the First Affiliated Hospital of Guizhou University of Traditional Chinese Medicine,Guiyang 550001,Guizhou Province,China)

机构地区：[1]贵州中医药大学第一附属医院眼科,贵州省贵阳市550001

出　　处：《眼科新进展》2025年第5期348-353,共6页Recent Advances in Ophthalmology

基　　金：贵州省中医药管理局中医药、民族医药科学技术研究课题(编号:QZYY-2019-005)。

摘　　要：目的探究山茱萸新苷对糖尿病视网膜病变(DR)大鼠的治疗效果,并分析活性氧(ROS)/硫氧还蛋白相互作用蛋白(TXNIP)/NOD样受体热蛋白结构域相关蛋白3(NLRP3)信号通路是否是其作用机制。方法将造模成功的56只DR大鼠随机分成模型组、山茱萸新苷20 mg·kg^(-1)组、山茱萸新苷40 mg·kg^(-1)组、羟苯磺酸钙5.8 mg·kg^(-1)组4组,每组14只;另取14只健康大鼠作为对照组。各组给予相应干预后,ELISA法检测视网膜组织炎症及氧化应激指标和血清空腹血糖(FBG)、血管生成因子水平;HE染色、TUNEL染色分别观察视网膜组织病理及视网膜细胞凋亡情况;荧光定量PCR法检测视网膜组织ROS及TXNIP、NLRP3 mRNA表达;蛋白印迹法检测视网膜细胞凋亡和ROS/TXNIP/NLRP3信号通路相关蛋白表达。结果与对照组比较,模型组大鼠视网膜细胞排列紊乱,数量明显减少,出现核固缩和水肿现象,白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、丙二醛(MDA)、FBG、血管生成素-1(Ang-1)、血管内皮生长因子(VEGF)、视网膜细胞凋亡率、Bax蛋白及ROS、TXNIP、NLRP3 mRNA和蛋白表达水平均显著升高,超氧化物歧化酶(SOD)、Bcl-2蛋白表达水平均显著降低(均为P<0.05)。与模型组比较,山茱萸新苷20 mg·kg^(-1)组、山茱萸新苷40 mg·kg^(-1)组、羟苯磺酸钙5.8 mg·kg^(-1)组大鼠视网膜细胞排列逐渐正常,数量增多,核固缩和水肿现象缓解,IL-6、TNF-α、MDA、FBG、Ang-1、VEGF、视网膜细胞凋亡率、Bax蛋白及ROS、TXNIP、NLRP3 mRNA和蛋白表达水平均显著降低,SOD、Bcl-2蛋白表达水平均显著升高(均为P<0.05)。DR大鼠视网膜组织病理损伤及上述定量指标的改善程度组间对比,山茱萸新苷40 mg·kg^(-1)组优于山茱萸新苷20 mg·kg^(-1)组(均为P<0.05);羟苯磺酸钙5.8 mg·kg^(-1)组优于山茱萸新苷20 mg·kg^(-1)组,但不如山茱萸新苷40 mg·kg^(-1)组(均为P<0.05)。结论山茱萸新苷能够改善DR大鼠视网膜炎Objective To explore the therapeutic effect of cornuside on diabetic retinopathy(DR)in rats and analyze the acting mechanism of the reactive oxygen species(ROS)/thioredoxin interacting protein(TXNIP)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)signaling pathway in this process.Methods A total of 56 successfully modeled DR rats were randomly divided into the model group,the cornuside 20 mg·kg^(-1)group,the cornuside 40 mg·kg^(-1)group,the calcium dobesilate 5.8 mg·kg^(-1)group,with 14 rats in each group;while,meanwhile another 14 healthy rats were selected as the control group.After the corresponding intervention of rats in each group,retinal tissue inflammation,oxidative stress indicators,fasting blood glucose(FBG),and angiogenic factor levels were detected by the enzyme-linked immunosorbent assay(ELISA).The hematoxylin-eosin(HE)staining and terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)staining were used to observe retinal histopathology and retinal cell apoptosis,respectively.The mRNA expression of ROS,TXNIP,and NLRP3 in the retinal tissue was detected by the quantitative fluorescence polymerase chain reaction(PCR).The apoptosis of retinal cells and the protein expression of the ROS/TXNIP/NLRP3 signaling pathway were detected by Western blotting.Results Compared with the control group,the model group showed disordered arrangement of retinal cells and a significant decrease in the cell number,accompanied by nuclear condensation and edema;interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),malondialdehyde(MDA),FBG,angiopoietin-1(Ang-1),vascular endothelial growth factor(VEGF),retinal cell apoptosis rate,ROS,and the mRNA and protein expression levels of TXNIP and NLRP3 increased significantly,while superoxide dismutase(SOD)and B cell lymphoma-2(Bcl-2)protein expression levels decreased significantly(all P<0.05).Compared with the model group,the arrangement of retinal cells in the cornuside 20 mg·kg^(-1)group,the cornuside 40 mg·kg^(-1)group,and the calcium dobesil

关 键 词：山茱萸新苷 糖尿病视网膜病变 视网膜细胞凋亡 活性氧 硫氧还蛋白相互作用蛋白 NOD样受体热蛋白结构域相关蛋白3 

分 类 号：R774.1[医药卫生—眼科]