摘要
目的建立视网膜缺血-再灌注损伤(retinal ischemia reperfusion injury,RIRI)模型,观察坏死性凋亡是否参与其病理过程。方法野生型C57小鼠随机分为对照组及实验组。对照组不做任何处理,实验组采用前房灌注法建立RIRI模型,并于RIRI后1 d、2d、4 d、7 d分别收集视网膜或眼球,行荧光定量PCR、Western blot、免疫荧光染色检测受体相互作用蛋白(receptor interacting protein,RIP)3及RIP1、白细胞介素-1β、白细胞介素-6、肿瘤坏死因子-α、Caspase8的表达变化。结果荧光定量PCR检测RIP3、RIP1、白细胞介素-1β、白细胞介素-6、肿瘤坏死因子-α、Caspase8的mRNA表达,与对照组比较,实验组在不同时间点均有显著升高,差异均有统计学意义(均为P<0.001)。RIP3及RIP1表达以早期升高为主,后期逐渐下降。Western blotting检测发现RIP3在RIRI后1 d及2 d显著升高,随后呈下降趋势。组织切片免疫荧光染色也发现RIP3在RIRI后1 d及2 d显著升高,随后呈下降趋势。结论实验性RIRI模型中,坏死性凋亡参与了其病理过程。坏死性凋亡特异性蛋白RIP3表达以早期升高为主,后期逐渐下降。
Objective To observe whether necroptosis involved in the pathologi- cal process of retinal ischemia reperfusion injury （RIRI） or not by establishing an reti- nal ischemia reperfusion injury model. Methods Wild type C57 mice were divided in- to control group and experimental group randomly. The mice in control group were treated with nothing. RIRI models in experimental group were induced with anterior chamber perfusion. The retina or eyeballs were harvested, Q-PCR, Western blot and im- munofluorescence staining were performed to test receptor interacting protein 3 （ RIP3 ）, RIP1, IL-1β, TNF-α, Caspase-8, and IL-5 changes at 1 day, 2 days, 4 days, 7 days. Results The mRNA expression change of RIP3, RIP1, IL-1β, TNF-α, Caspase-8, and IL-6 were detected by Q-PCR. Compared with control group, all genes mRNA ex- pression were increased significantly at different time points （ all P 〈0.001 ）. RIP3 ,RIP1 mRNA expressions were increased at the early phase of RIRI, and decreased in the late phase slowly. Western blot found RIP3 expression increased dramatically at 1 day and 2 days, then decreased gradually. Retinal immunofluorescence staining also found RIP3 expression increased dramatically at 1 day and 2 days ,then decreased slowly. Conclu- sion Necroptosis participates in the pathological process of experimental RIRI. As a specified marker of necroptosis, RIP3 expression increase at the early phase of RIRI, and decrease at the later phase gradually.
出处
《眼科新进展》
CAS
北大核心
2016年第12期1109-1112,共4页
Recent Advances in Ophthalmology
基金
国家自然科学基金资助(编号:81560168)~~
