清化瘀热方通过IL-33/ST2通路缓解糖尿病视网膜病变大鼠中视网膜细胞炎症反应  

Qinghua Yure Fang Alleviates Inflammatory Response in Retinal Cells of Diabetic Retinopathy Rats via IL-33/ST2 Pathway

作　　者：翟丽萍[1] 王旭 王谦 陆骏[2] ZHAI Li-ping;WANG Xu;WANG Qian;LU Jun(Department of Endocrinology,Taizhou Traditional Chinese Medicine Hospital,Taizhou 225300,China;Department of Ophthalmology,Taizhou Traditional Chinese Medicine Hospital,Taizhou 225300,China)

机构地区：[1]泰州市中医院内分泌科,江苏泰州225300 [2]泰州市中医院眼科,江苏泰州225300

出　　处：《南昌大学学报（医学版）》2024年第6期33-41,F0003,共10页Journal of Nanchang University：Medical Sciences

基　　金：江苏省中医药科技发展计划项目(MS2021085);泰州市科技项目(TS202011)。

摘　　要：目的探讨清化瘀热方(Qinghua Yure Fang,QHYRF)对糖尿病视网膜病变(diabetic retinopathy,DR)大鼠中视网膜细胞炎症、凋亡和氧化应激的影响及信号网络机制。方法通过链脲佐菌素(streptozotocin,STZ)注射建立DR大鼠模型和人视网膜微血管内皮细胞(human retinal microvascular endothelial cells,hRMECs)高糖处理的细胞模型。使用苏木精-伊红(hematoxylin-eosin,HE)染色和透射电镜观察视网膜组织。使用相应试剂盒、酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)和蛋白质免疫印迹分别对hRMEC中氧化应激指标、促炎细胞因子和促凋亡蛋白的表达模式进行表征。蛋白质免疫印迹法分析白细胞介素33(interleukin-33,IL-33)/生长刺激表达基因2(suppression of tumorigenicity 2,ST2)信号通路相关蛋白的表达。结果HE染色结果显示,与假手术大鼠相比,STZ处理的大鼠表现出明显的水肿、毛细血管壁增厚、内皮细胞增生和纤维组织增生,并通过以剂量依赖的方式给予QHRYF治疗后得到部分缓解(P<0.05)。视网膜的超微结构观察结果显示,与假手术大鼠相比,STZ处理大鼠的毛细血管基底膜厚度(basement membrane thickness,BMT)增加,而QHRYF治疗以剂量依赖的方式降低了BMT值(P<0.05)。与假手术大鼠相比,STZ处理的大鼠IL-33和ST2的表达显著降低(P<0.05),QHRYF处理以剂量依赖的方式提高了IL-33和ST2的表达(P<0.05)。与假手术大鼠相比,STZ大鼠视网膜组织中的活性氧(reactive oxygen species,ROS)和3,4-亚甲二氧亚氨基苯丙胺(3,4-methylenedioxyamphetamine,MDA)水平显著升高、超氧化物歧化酶(superoxide dismutase,SOD)水平降低(P<0.05),而这些变化均通过QHRYF以剂量依赖的方式部分逆转(P<0.05)。与假手术大鼠相比,STZ处理的大鼠细胞上清液中白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)和C反应蛋白(C-reactive protein,CRP)的含量显著增加。与注射磷酸缓冲�Objective To investigate the effects of Qinghua Yure Fang(QHYRF)on inflammation,apoptosis and oxidative stress in retinal cells of diabetic retinopathy(DR)rats and the mechanism of signaling network.Methods A DR rat model and a cellular model of human retinal microvascular endothelial cells(hRMECs)treated with high glucose were established by streptozotocin(STZ)injection.Retinal tissue was observed using hematoxylin-eosin(HE)staining and transmission electron microscopy.The expression patterns of oxidative stress indexes,pro-inflammatory cytokines and pro-apoptotic proteins in hRMEC were characterized using corresponding kits,enzyme linked immunosorbent assay(ELISA)and western blot.The expression of proteins associated with Interleukin-33(IL-33)/suppression of Tumorigenicity 2(ST2)signaling pathway was analyzed by Western blot.Results HE staining results showed that STZ-treated rats exhibited significant edema,capillary wall thickening,endothelial cell hyperplasia,and fibrous tissue hyperplasia compared with sham-operated rats,all of which could be partially alleviated by QHRYF treatment given in a dose-dependent manner(P<0.05).Observation of the ultrastructure of the retina showed that STZ-treated rats had increased capillary basement membrane thickness(BMT)values compared with sham-operated rats,while QHRYF treatment decreased BMT values in a dose-dependent manner(P<0.05).The expression of IL-33 and ST2 was significantly reduced in STZ-treated rats compared with sham-operated rats,and QHRYF treatment increased the expression of IL-33 and ST2 in a dose-dependent manner(P<0.05).Compared with sham-operated rats,STZ rats showed significantly higher levels of reactive oxygen species(ROS)and MDA and lower levels of SOD in retinal tissues(P<0.05),all of which were partially reversed by QHRYF in a dose-dependent manner(P<0.05).The levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and C-reactive protein(CRP)were significantly increased in cell supernatants of STZ-treated rats,compared with sham-operated rat

关 键 词：清化瘀热方 IL-33/ST2信号通路 糖尿病视网膜病变 炎症 细胞凋亡 动物实验 大鼠 

分 类 号：R-332[医药卫生]