虎杖苷激活MEK/ERK信号通路改善东莨菪碱诱导的干眼症大鼠眼表功能障碍和病变  

Polydatin activates MEK/ERK signaling pathway to improve ocular surface dysfunction and pathology in rats with scopolamine-induced dry eye disease

作　　者：曹瑾 龚兰兰 吕旭东 CAO Jin;GONG Lanlan;LYU Xudong(Department of Ophthalmology,Xianning Central Hospital,Xianning 437000,China)

机构地区：[1]咸宁市中心医院眼科,咸宁437000

出　　处：《中国免疫学杂志》2024年第5期1089-1095,共7页Chinese Journal of Immunology

基　　金：咸宁市中心医院湖北科技学院附属第一医院科研项目(2021XYB017)。

摘　　要：目的:探究虎杖苷对干眼症(DED)大鼠眼表功能障碍和病变的影响及其保护机制。方法:90只SD大鼠随机选取18只为对照组,其余皮下注射氢溴酸东莨菪碱建立DED模型,建模后分为模型组、0.05%虎杖苷组、0.5%虎杖苷组、0.5%虎杖苷+U0126[丝裂原活化蛋白激酶激酶(MEK)抑制剂]组,每组18只;各组给予相应剂量药物进行干预,对照组和模型组大鼠给予等量生理盐水。干预第7、14、21、28天,进行Schirmer测试检测各组大鼠泪液分泌量,荧光素染色检测各组大鼠泪膜破裂时间(BUT)和角膜损伤;RT-qPCR检测角膜和结膜组织促炎因子TNF-α、IFN-γ和IL-1βmRNA表达;HE染色观察角膜组织病理学变化;PAS染色评估结膜杯状细胞数量;Western blot检测角膜和结膜组织MEK/细胞外调节蛋白激酶(ERK)通路相关蛋白表达。结果:与对照组相比,模型组大鼠泪液分泌量、BUT、结膜杯状细胞数量显著减少(P<0.05),角膜损伤评分、TNF-α、IFN-γ和IL-1βmRNA水平显著升高(P<0.05),角膜上皮层变薄,可见大量炎症细胞浸润和新生毛细血管;与模型组相比,0.05%虎杖苷组和0.5%虎杖苷组泪液分泌量、BUT、结膜杯状细胞数量、p-MEK1/2/MEK1/2和p-ERK1/2/ERK1/2显著升高(P<0.05),角膜损伤评分、TNF-α、IFN-γ和IL-1βmRNA水平显著降低(P<0.05),角膜组织病理损伤明显缓解;U0126可显著减弱虎杖苷对DED大鼠眼表功能障碍和病变的保护作用。结论:虎杖苷可能通过激活MEK/ERK通路改善东莨菪碱诱导的DED大鼠眼表功能障碍和病变。Objective:To explore effect of polydatin on ocular surface dysfunction and pathology in dry eye disease(DED)rats and its protective mechanism.Methods:Eighteen from 90 SD rats were randomly chosen as control group,and other rats were injected subcutaneously with scopolamine hydrobromide to establish DED model.After modeling,rats were separated into model group,0.05%polydatin group,0.5%polydatin group,0.5%polydatin+U0126[mitogen-activated protein kinase kinase(MEK)inhibitor]group,with 18 rats in each group;each administration group was given corresponding doses of drugs for intervention,and rats in control group and model group were given same amount of normal saline.On the 7th,14th,21st,and 28th day of intervention,Schirmer test was performed to measure tear secretion amount of rats in each group,fluorescein staining was performed to measure tear film breakup time(BUT)and corneal damage of rats in each group;RT-qPCR was performed to measure mRNA expression of proinflammatory factors TNF-α,IFN-γand IL-1βin cornea and conjunctival tissues;HE staining was performed to observe histopathological changes of cornea;PAS staining was performed to evaluate number of conjunctival goblet cells;Western blot was performed to detect expressions of MEK/extracellular regulatory protein kinase(ERK)pathway related proteins.Results:Compared with control group,tear secretion amount BUT and number of conjunctival goblet cells in model group were obviously reduced(P<0.05),corneal injury score,TNF-α,IFN-γand IL-1βmRNA levels were obviously increased(P<0.05),corneal epithelial layer was thinned,a large number of inflammatory cell infiltration and new capillaries could be seen;compared with model group,tear secretion amount,BUT,number of conjunctival goblet cells,p-MEK1/2/MEK1/2 and p-ERK1/2/ERK1/2 in 0.05%polydatin group and 0.5%polydatin group were obviously increased(P<0.05),corneal injury score,TNF-α,IFN-γand IL-1βmRNA levels were obviously reduced(P<0.05),pathological damage of corneal tissue was obviously relieved;U0126 coul

关 键 词：干眼症 虎杖苷 炎症 眼表功能障碍 丝裂原活化蛋白激酶激酶 细胞外调节蛋白激酶 

分 类 号：R777.34[医药卫生—眼科]