摘要
目的观察N-乙酰-5-羟色胺(NAS)对视网膜缺血再灌注损伤(RIRI)大鼠视网膜中肿瘤坏死因子(TNF)-α蛋白表达的影响,探讨其可能机制。方法采用随机数字表法将90只健康雄性Sprague-Dawley大鼠分为假手术组(10只)、RIRI组(40只)、NAS组(40只)。以右眼为实验眼。RIRI组、NAS组大鼠采用前房高眼压法建立RIRI模型。NAS组大鼠分别于建模前及建模后30 min腹腔注射10 mg/kg NAS。建模后6、12、24、72 h,苏木精-伊红染色观察各组大鼠视网膜组织病理变化,计数视网膜神经节细胞(RGC);免疫组织化学染色、蛋白质免疫印迹法(Western blot)检测各组大鼠视网膜中TNF-α、核因子E2相关因子2(Nrf2)、血红素氧合酶-1(HO-1)蛋白相对表达量。组间差异采用单因素方差分析。采用一般线性回归法分析NAS干预后TNF-α蛋白相对表达量变化与Nrf2、HO-1蛋白相对表达量变化的相关性。结果光学显微镜观察发现,建模后6、12、24 h,RIRI组大鼠视网膜水肿;NAS组大鼠视网膜厚度显著薄于RIRI组,差异有统计学意义(F=9.645、477.150、2.432,P<0.01)。建模后6、12、24、72 h,NAS组大鼠视网膜RGC计数显著高于RIRI组,差异有统计学意义(F=12.225、12.848、117.655、306.394,P<0.05)。免疫组织化学染色、Western blot检测结果显示,建模后6 h,RIRI组大鼠视网膜中TNF-α蛋白相对表达量较假手术组显著增加,12 h达较高水平,24、72 h下降,但均显著高于假手术组,差异有统计学意义(免疫组织化学染色:F=105.893、1356.076、434.026、337.351,P<0.01;Western blot:F=92.906、534.948、327.600、385.324,P<0.01);建模后不同时间点,NAS组大鼠视网膜中TNF-α蛋白相对表达量显著低于RIRI组(免疫组织化学染色:F=15.408、570.482、21.070、13.767,P<0.05;Western blot:F=12.618、115.735、13.176、111.108,P<0.05),但仍高于假手术组(免疫组织化学染色:F=40.709、151.032、156.321、216.035,P<0.01;Western blot:F=33.943、79.729、74.057、64.488,P<
Objective To dynamically observe the effect of N-acetylserotonin(NAS)on the expression of tumor necrosis factor-α(TNF-α)protein in retina of retinal ischemia reperfusion injury(RIRI)rats,and to explore the mechanism.Methods By using random number table method,90 healthy male Sprague-Dawley rats were divided into sham operation group(n=10),RIRI group(n=40),and NAS group(n=40).The right eye was as the experimental eye.In the RIRI group and NAS group,the anterior chamber high intraocular pressure method was used to establish the RIRI model.In the NAS group,10 mg/kg NAS was injected intraperitoneally before modeling and 30 minutes after modeling.At 6,12,24,72 h after modeling,hematoxylin-eosin staining was used to observe the pathological changes of the retina,and the retinal ganglion cells(RGC)were counted.Each group was detected by immunohistochemical staining and Western blot about the relative expression of TNF-α,nuclear factor E2-related factor 2(Nrf2),and heme oxygenase-1(HO-1)protein in the rat retina.Oneway analysis of variance was used for differences between groups.The general linear regression method was used to analyze the correlation between the relative expression changes of TNF-αprotein and the changes of Nrf2 and HO-1 protein expression after NAS intervention.Results Optical microscope observation revealed that the retinal edema of rats in the RIRI group was observed at 6,12,and 24 h after modeling;the thickness of the retina in the NAS group was significantly thinner than that in the RIRI group,and the difference was statistically significant(F=9.645,477.150,2.432;P<0.01).At 6,12,24,and 72 h after modeling,the retinal RGC counts in the NAS group were significantly higher than those in the RIRI group,and the difference was statistically significant(F=12.225,12.848,117.655,306.394;P<0.05).The results of immunohistochemical staining and Western blot showed that 6 h after modeling,the relative expression of TNF-αprotein in the retina of the RIRI group increased significantly compared with that in th
作者
殷慧文
张宁
尹伊
张学宁
刘建晓
刘晓阳
王晓莉
赵岩松
Yin Huiwen;Zhang Ning;Yin Yi;Zhang Xuening;Liu Jianxiao;Liu Xiaoyang;Wang Xiaoli;Zhao Yansong(Teaching and Research Office of Ophthalmology,Weifang Medical University,Weifang 261053,China;School of Medical Imaging,Weifang Medical University,Weifang 261053,China;Department of Ophthalmology,The Affiliated Hospital of Weifang Medical University,Weifang 261031,China)
出处
《中华眼底病杂志》
CAS
CSCD
北大核心
2021年第6期462-469,共8页
Chinese Journal of Ocular Fundus Diseases
基金
国家自然科学基金(82071888)
山东省自然科学基金(ZR2020MH074)
山东省医药卫生发展计划项目(2017WS738)
山东省中医药科技发展计划项目(2019-0421)。
