基于丝裂原活化蛋白激酶信号通路灯盏细辛对大鼠急性高眼压的缓解及视网膜神经节细胞的保护作用  

Effect of erigeron breviscapus on relieving acute ocular hypertension and protecting retinal ganglion cells in glaucoma rats based on mitogen activated protein kinase signaling pathway

作　　者：冉文瑛 朱冬梅 王媛 Ran Wenying;Zhu Dongmei;Wang Yuan(Department of Ophthalmology,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450007,China)

机构地区：[1]郑州大学附属郑州中心医院眼科,450007

出　　处：《中华眼底病杂志》2021年第6期455-461,共7页Chinese Journal of Ocular Fundus Diseases

基　　金：河南省医学科技攻关计划项目(2018020799);河南省高等学校重点科研项目(19A320076)。

摘　　要：目的观察灯盏细辛(EBHM)调控丝裂原活化蛋白激酶(MAPK)信号通路对大鼠急性高眼压的影响及视网膜神经节细胞(RGC)的保护作用。方法雄性Sprague-Dawley大鼠60只,采用随机数字表法分为对照组、模型组、EBHM低剂量组(A组)、EBHM中剂量组(B组)、EBHM高剂量组(C组),每组均为12只;以左眼为实验眼。模型组、A组、B组、C组大鼠通过前房加压灌注生理盐水构建急性高眼压模型;对照组仅给予全身麻醉。建模后2~30 d,对照组、模型组大鼠灌胃生理盐水3 ml,1次/d;A组、B组、C组大鼠分别给予0.30、0.45、0.60 g/100 g EBHM灌胃,1次/d。建模前以及建模后1、14、30 d测量大鼠眼压,并统计高眼压模型比例。建模后30 d,苏木精-伊红(HE)染色观察视网膜组织病理变化;免疫荧光染色检测RGC数量变化;实时荧光定量聚合酶链反应(RT-qPCR)检测各组大鼠视网膜中p38MAPK、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)mRNA相对表达量;蛋白质免疫印迹法(Western blot)检测各组大鼠视网膜中p38MAPK、磷酸化p38MAPK(p-p38MAPK)、Caspase-3蛋白相对表达量。多样本间比较采用单因素方差分析;两两样本间比较采用SNK-q检验。结果建模后1 d,对照组大鼠均未出现急性高眼压,其余各组均建模成功。与模型组比较,B组、C组建模后14 d急性高眼压率较低,差异有统计学意义(χ^(2)=98.701,P<0.05);A组、B组、C组建模后30 d急性高眼压率均为0。A组、B组、C组建模后14、30 d之间急性高眼压率比较,差异均无统计学意义(P>0.05)。HE染色结果显示,对照组大鼠视网膜结构完整,各层清晰可见;RGC计数未见异常,形态饱满圆润。模型组大鼠视网膜明显变薄;RGC数量大量减少,形态空泡化,排列稀疏。A组、B组、C组大鼠视网膜变厚,RGC数量增多,其中C组视网膜结构恢复程度更好。免疫荧光染色结果显示,A组、B组、C组大鼠RGC计数较模型组升高,差异有统计学意义(F=297.514,P<0.0Objective To investigate the effect of erigeron breviscapus(EBHM)on ocular hypertension and the protective effect of retinal ganglion cells(RGCs)in rats by regulating mitogen activated protein kinase(MAPK)signaling pathway.Methods Sixty male Sprague-Dawley rats were divided into control group,model group,low-dose EBHM group(group A),medium-dose EBHM group(group B),and high-dose EBHM group(group C)by random number table method.There were 12 rats in the group,the left eye was used as the experimental eye.The rats of model group,group A,group B,and group C were infused with normal saline through the anterior chamber to construct an acute ocular hypertension model;the control group was given general anesthesia only.Then,2-30 days after modeling,rats in the control group and model group were given 3 ml of normal saline once a day;rats in group A,group B,and group C were given 0.30,0.45,and 0.60 g/100 g EBHM by intragastric administration,respectively,1 time/d.The rat intraocular pressure was measured before modeling and 1,14,and 30 days after modeling,and the proportion of high intraocular pressure model was measured.Thirty days after modeling,hematoxylin-eosin(HE)staining was used to observe the pathological changes of retinal tissue;immunofluorescence staining was used to detect the changes in the number of RGCs;real-time fluorescent quantitative polymerase chain reaction(RT-qPCR)was used to detect p38 in the retinas of rats in each group.The relative expression of MAPK and Caspase-3 mRNA;western blot was used to detect p38MAPK and phosphorylation in the retina of rats in each group relative expression of phosphorylate-p38MAPK(p-p38MAPK)and Caspase-3 protein.One-way analysis of variance was used for multi-sample comparison,and SNK-q test was used for comparison between two samples.Results One day after modeling,none of the rats in the control group developed acute ocular hypertension,and the other groups were successfully modeled.Compared with the model group,the rates of acute ocular hypertension at 14 days after m

关 键 词：灯盏细辛 高眼压 P38丝裂原活化蛋白激酶类 视网膜神经节细胞 

分 类 号：R285.5[医药卫生—中药学]