丹蒌片通过Keap1-Nrf2/HO-1信号通路缓解视网膜缺血-再灌注损伤  

Danlou tablet alleviates retinal ischemia-reperfusion injury through Keap1-Nrf2/HO-1 pathway

作　　者：蔺彦娜 吴惠琴[2] 郑博[2] 陈晓冬 雷鹏 陈梦涵 Lin Yanna;Wu Huiqin;Zheng Bo;Chen Xiaodong;Lei Peng;Chen Menghan(Shaanxi University of Chinese Medicine,Xianyang 712046,Shaanxi Province,China;Seventh Department of Ophthalmology,Xi'an No.1 Hospital,Xi'an 710002,Shaanxi Province,China)

机构地区：[1]陕西中医药大学,陕西省咸阳市712046 [2]中国陕西省西安市第一医院眼七科,710002

出　　处：《国际眼科杂志》2024年第7期1027-1031,共5页International Eye Science

基　　金：陕西省中医药管理局科技项目基金(No.2021-ZZ-JC014)。

摘　　要：目的:探讨丹蒌片对小鼠视网膜缺血-再灌注损伤(RIRI)的保护作用及其机制。方法:将40只ApoE^(-/-)小鼠给予高脂饲料喂养6 wk,通过前房灌注加压法建立RIRI模型,分为对照组(给予生理盐水灌胃8 wk)、RIRI模型组(给予生理盐水灌胃8 wk)及丹蒌片低、中、高剂量组[分别给予丹蒌片溶液1、2、4 g/(kg·d)灌胃8 wk]。采用苏木精-伊红(HE)染色观察各组小鼠视网膜组织形态学变化情况,TUNEL染色检测各组小鼠视网膜细胞凋亡情况,Western-blot法检测各组小鼠视网膜组织中Kelch样环氧氯丙烷相关蛋白-1(Keap1)、转录因子核因子E2相关因子2(Nrf2)、血红素加氧酶-1(HO-1)、超氧化物歧化酶(Sod2)蛋白表达情况。结果:与对照组比较,RIRI模型组小鼠视网膜萎缩,厚度变薄,细胞凋亡增加,视网膜组织中Sod2蛋白表达下调,Keap1蛋白表达上调(均P<0.01);与RIRI模型组比较,丹蒌片中、高剂量组小鼠视网膜厚度增加(均P<0.01),丹蒌片低、中、高剂量组小鼠视网膜细胞凋亡降低(均P<0.05),丹蒌片低剂量组小鼠视网膜组织中Keap1和HO-1蛋白表达水平无明显差异(P>0.05),丹蒌片中、高剂量组小鼠视网膜组织中Sod2、Nrf2、HO-1蛋白表达上调,Keap1蛋白表达下调(均P<0.05)。结论:丹蒌片能缓解RIRI小鼠模型视网膜萎缩变薄,减少视网膜细胞凋亡,其作用是通过调控Keap1-Nrf2/HO-1信号通路降低RIRI氧化应激反应来实现。AIM:To investigate the protective effect and mechanism of Danlou tablet on retinal ischemia-reperfusion injury(RIRI)in mice.METHODS:A total of 40 ApoE^(-/-)mice were fed with high fat diet for 6 wk,and the RIRI model was established by anterior chamber infusion of pressurized saline.The mice were divided into control group(normal saline for 8 wk),RIRI model group(normal saline for 8 wk),and low-,medium-,and high-dose Danlou tablets groups[1,2,and 4 g/(kg·d),respectively,for 8 wk].The morphological changes of retina were observed by hematoxylin-eosin(HE)staining,retinal cell apoptosis was detected by terminal-deoxynucleoitidyl transferase mediated Nick-End Labeling(TUNEL)staining.The Western-blot assay was used to detect the expression of retinal tissue sample Kelch-like ech-associated protein 1(Keap1),nuclear factor E2 related factor 2(Nrf2),heme oxygenase 1(HO-1),and superoxide dismutase(Sod2)proteins.RESULTS:Compared with the control group,the mouse retina was atrophic with thinning thickness and increasing cell apoptosis,down-regulation of Sod2 protein expression,and up-regulation of Keap1 protein expression in the RIRI model group(all P<0.01).Compared with the RIRI model group,the retinal thickness increased in the medium-and high-dose of Danlou tablets groups(all P<0.01),and the cell apoptosis of retina decreased in the low-,medium-and high-dose of Danlou tablets groups(all P<0.05).There were no significant differences in the expression of Keap1 and HO-1 proteins of mouse retina tissue in the low-dose of Danlou tablets group(P>0.05).The expression of Sod2,Nrf2 and HO-1 proteins up regulated,and the expression of Keap1 protein down regulated in the medium-and high-dose of Danlou tablets groups(all P<0.05).CONCLUSION:Danlou tablet can alleviate RIRI-induced atrophy and thinning of retina and retinal cell apoptosis by regulating Keap1-Nrf2/HO-1 signal pathway and reducing oxidative stress.

关 键 词：丹蒌片 ApoE^(-/-)小鼠 高脂血症 视网膜缺血-再灌注损伤 Keap1-Nrf2/HO-1通路 

分 类 号：R285.5[医药卫生—中药学]