基于PI3K/AKT信号通路探讨Netrin-1对糖尿病性白内障大鼠晶状体上皮细胞的保护作用  

To explore the protective effect of Netrin-1 on the lens epithelial cells of rats with diabetic cataract based on PI3K/AKT signaling pathway

作　　者：周米露 赵锐[2] 王大庆[2] 曾晓莉 ZHOU Mi-lu;ZHAO Rui;WANG Da-qing;ZENG Xiao-li(Department of Ophthalmology,Sichuan Nanchong Central Hospital,Nanchong,Sichuan Province 637000,China)

机构地区：[1]四川省南充市中心医院眼科,四川南充637000 [2]四川省南充市川北医学院,四川南充637000

出　　处：《解剖学研究》2021年第6期577-583,共7页Anatomy Research

基　　金：四川省医学科研课题计划(S19055)。

摘　　要：目的基于磷脂酰肌醇3激酶/蛋白激酶B(PI3K/AKT)信号通路探讨Netrin-1对糖尿病性白内障(DC)大鼠晶状体上皮细胞(LEC)的保护作用。方法将80只大鼠随机分为对照组(Control组)、模型组(Vehicle组)、治疗组(Netrin-1组)、PI3K/AKT信号通路干预组(LY294002组)。采用腹腔注射链尿佐菌素(STZ)制作大鼠DC模型,Netrin-1组玻璃体腔注射Netrin-1,LY294002组注射Netrin-1+LY294002,Control组、Vehicle组注射等体积PBS,干预1个月。采用苏木精-伊红(HE)染色观察大鼠晶状体组织病理学改变;免疫组化染色观察晶状体组织E-钙黏蛋白(E-cadherin)、基质金属蛋白酶2(MMP-2)的表达;检测晶状体组织氧化应激指标超氧化物歧化酶(SOD)、丙二醛(MDA)含量;MTT法检测LEC增殖活性;Hochest 33258染色检测LEC凋亡情况;透射电镜观察各组LEC超微结构的变化;Western blot检测晶状体组织p-PI3K、p-AKT、caspase-3蛋白表达。结果与Control组相比,Vehicle组大鼠晶状体变浑浊,条索状、片状的LEC向晶状体成纤维细胞聚集,LEC增殖活力明显降低,凋亡能力明显增加,超微结构损伤明显,E-cadherin、SOD、p-PI3K、p-AKT表达减少,MMP-2、MDA、caspase-3表达增多(P<0.05);与Vehicle组相比,Netrin-1组大鼠晶状体浑浊程度得到明显改善,可见少量LEC向晶状体纤维细胞迁移,增殖活力改善,凋亡细胞减少,细胞形态逐渐改善,E-cadherin、SOD、p-PI3K、p-AKT表达增多,MMP-2、MDA、caspase-3表达减少(P<0.05)。LY294002组晶状体损伤程度明显增加,与Vehicle组差异无统计学意义(P>0.05)。结论Netrin-1通过激活PI3K/AKT信号通路,保护晶状体组织免受高血糖诱导的氧化应激和细胞凋亡,抑制LEC的迁移和EMT,从而起到延缓糖尿病大鼠白内障进展的作用。Objective Based on Phosphatidylinositol 3-kinase/Protein kinase B(PI3K/AKT)signaling pathway to explore the effect of Netrin-1 on diabetic cataract(DC)The protective effect of mouse lens epithelium cell(LEC).Methods 80 rats were randomly divided into control group(Control group),model group(Vehicle group),treatment group(Netrin-1 group),PI3K/AKT signal pathway intervention group(LY294002 group).The rat DC model was made by intraperitoneal injection of streptozotocin(STZ),the Netrin-1 group was injected with Netrin-1 into the vitreous cavity,the LY294002 group was injected with Netrin-1+LY294002,the Control group and the Vehicle group were injected with equal volume of PBS,intervention 1 month.HE staining was used to observe the histopathological changes of rat lens;immunohistochemistry was used to observe the expression of E-cadherin and matrix metalloproteinase 2(MMP-2)in lens tissue;to detect the oxidative stress of lens tissue.The content of superoxide dismutase(SOD)and malondialdehyde(MDA);MTT method to detect LEC proliferation activity;Hochest 33258 staining to detect LEC apoptosis;transmission electron microscopy to observe the ultrastructure of LEC Changes;Western blot detection of p-PI3K,p-AKT,and caspase-3 protein expression in lens tissue.Results Compared with the control group,the lens of the vehicle group became turbid,the cord-like and sheet-like LECs aggregated to the lens fibroblasts,the proliferation of LECs was significantly reduced,the apoptotic capacity was significantly increased,and the ultrastructure damage was obvious.The expression of cadherin,SOD,p-PI3K,and p-AKT decreased,and the expression of MMP-2,MDA,and caspase-3 increased(P<0.05).Compared with the vehicle group,the lens opacity of the rats in the Netrin-1 group was significantly improved,and a small amount of LEC was seen to the lens Fibroblast migration,improved proliferation activity,decreased apoptotic cells,gradually improved cell morphology,increased expression of E-cadherin,SOD,p-PI3K,and p-AKT,and decreased expression of MMP-

关 键 词：糖尿病性白内障 磷脂酰肌醇3激酶/蛋白激酶B NETRIN-1 晶状体上皮细胞 

分 类 号：R587.2[医药卫生—内分泌] R776.1[医药卫生—内科学]