α1-抗胰蛋白酶联合骨髓间充质干细胞通过p38 MAPK/NF-κB信号通路调节糖尿病大鼠视网膜病变的研究  

α1-antitrypsin combined with bone marrow mesenchymal stem cells regulates retinopathy in diabetic rats via p38 MAPK/NF-κB signaling pathway

作　　者：陈红 李春花[1] 王文军[1] 曾戎[1] 闫欢欢 张宏[2] CHEN Hong;LI Chun-hua;WANG Wen-jun;ZENG Rong;YAN Huan-huan;ZHANG Hong(Department of Ophthalmology, Xi'an Fourth Hospital, Xi'an, Shaanxi 710004, China;School of Basic Medicine,Xi'an Jiaotong University,Xi'an,Shaanxi 710049,China)

机构地区：[1]西安市第四医院眼科,陕西西安710004 [2]西安交通大学基础医学院,陕西西安710049

出　　处：《海南医学院学报》2021年第1期11-17,共7页Journal of Hainan Medical University

基　　金：陕西省重点研发项目(2019SF-084)。

摘　　要：目的:探讨α1-抗胰蛋白酶联合骨髓间充质干细胞(BMSC)对糖尿病大鼠视网膜病变的影响及作用机制。方法:通过注射链脲佐菌素法构建糖尿病视网膜病变模型,30只造模成功的大鼠随机分为模型组、BMSC组、联合组(α1-抗胰蛋白酶联合BMSC),处理后4周,测定糖尿病大鼠血糖、血清胰岛素水平,ELISA测定血清中IL-1β、IL-6和TNF-α含量,HE检测视网膜病理形态,TUNEL染色检测视网膜神经细胞凋亡情况,免疫组化染色检测视网膜组织CD45的表达水平,实时荧光定量PCR检测视网膜VEGF、HIF-1α及ANGⅡmRNA的表达,Western blot法检测视网膜组织中p38 MAPK/NF-κB信号通路相关蛋白表达。结果:与对照组比较,模型组大鼠血糖增加,胰岛素水平下降,血清中IL-1β、IL-6和TNF-α含量升高,视网膜出现明显病变,各层细胞排列杂乱,视网膜神经细胞凋亡较多,视网膜组织中CD45呈高表达,VEGF、HIF-1α、ANGⅡmRNA表达升高,p-p38、p-p65、p-IκBα蛋白表达升高(P均<0.05)。与模型组比较,BMSC组和联合组血糖有所下降,胰岛素水平增加,血清中IL-1β、IL-6和TNF-α含量下降,视网膜病变得以改善,视网膜神经细胞凋亡减少,视网膜组织CD45表达降低,VEGF、HIF-1α、ANGⅡmRNA表达下降,p-p38、p-p65、p-IκBα蛋白表达下降,且与BMSC组比较,联合组的作用效果更明显(P均<0.05)。结论:α1-抗胰蛋白酶联合BMSC移植可改善糖尿病大鼠的视网膜病变程度,机制可能与抑制p38 MAPK/NF-κB信号通路有关。Objective:To investigate the effect ofα1-antitrypsin combined with bone marrow mesenchymal stem cells on retinopathy in diabetic rats and its mechanism.Methods:A model of diabetic retinopathy was established by intraperitoneal injection of streptozotocin.The 30 Wistar rats successfully modeled were randomly divided into a model group,a bone marrow mesenchymal stem cell group and a combined group(α1-antitrypsin combined with bone marrow Mesenchymal stem cells);the blood glucose and serum insulin levels of diabetic rats were measured 4 weeks after treatment.Enzyme-linked immunosorbent assay(ELISA)was for measuring serum inflammatory factors IL-1β,IL-6 and TNF-αin rats.Observing the pathological morphology of rat retina was under hematoxylin-eosin staining(HE).TUNEL staining was to observe the apoptosis of rat retinal nerve cells.Immunohistochemical method was used to detect the expression level of CD45 in retinal tissue.Real-time fluorescence quantitative PCR was used to detect the expression of retinal vascular endothelial growth factor(VEGF),hypoxia-inducible factor-1α(HIF-1α),and angiotensinⅡ(ANGⅡ)mRNA.Western blot was used to detect the expression of p38 MAPK/NF-κB signaling pathway-related proteins in the retinal tissue of each group of rats.Results:Compared with the control group,the rats in the model group had increased blood glucose,decreased insulin levels,increased serum IL-1β,IL-6,and TNF-αlevels,and had obvious lesions in the retina.CD45 showed high expression in retinal tissue,VEGF,HIF-1α,ANGⅡmRNA expression increased,p-p38,p-p65,p-IκBαprotein expression increased(P<0.05).Compared with the model group,the bone marrow mesenchymal stem cell group and the combined group have decreased blood glucose,increased insulin levels,and decreased serum IL-1β,IL-6 and TNF-αlevels.Retinopathy was improved,apoptosis of retinal nerve cells was reduced,CD45 expression in retinal tissue was reduced,VEGF,HIF-1α,ANGⅡmRNA expression was decreased,and p-p38,p-p65,p-IκBαprotein expression was decrease

关 键 词：Α1-抗胰蛋白酶 骨髓间充质干细胞 糖尿病 视网膜病变 血管内皮生长因子 p38 MAPK/NF-κB通路 

分 类 号：R587.2[医药卫生—内分泌] R774.1[医药卫生—内科学]