中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
Chinese Journal of Experimental Ophthalmology
2015年
11期
985-990
,共6页
刘巾男%何宇%张军军%范玮
劉巾男%何宇%張軍軍%範瑋
류건남%하우%장군군%범위
视网膜缺血-再灌注%视网膜神经节细胞%肿瘤坏死因子-α%微小RNA-181a
視網膜缺血-再灌註%視網膜神經節細胞%腫瘤壞死因子-α%微小RNA-181a
시망막결혈-재관주%시망막신경절세포%종류배사인자-α%미소RNA-181a
Retinal ischemia-reperfusion%Retinal ganglion cells%Tumor necrosis factor-α%microRNA-181a
背景 视网膜缺血-再灌注(RIR)损伤是眼科常见病理改变之一,但其发病机制尚未明确.目的探讨大鼠RIR模型中微小RNA-181a(miR-181a)与视网膜神经节细胞(RGCs)凋亡之间的关系及可能的靶向机制.方法 构建68只SD大鼠RIR模型,按随机数字表法随机分为对照组和造模后即刻组、造模后24 h组及造模后72 h组,每组17只,根据MiRanda、Targetscan和miRBase 3个靶基因数据库的共同预测结果,选取肿瘤坏死因子-α(TNF-α)作为miR-181a的下游靶基因研究对象,通过免疫荧光标记法Western blot 及实时荧光定量PCR法观察miR-181a、TNF-α在各组的表达情况及其与RGCs凋亡的关系. 结果 RGCs计数在造模后24 h和72 h显著减少,与对照组比较差异有统计学意义(P＜0.001).各造模组miR-181a表达量随造模时间明显下降,与对照组比较差异有统计学意义(P＜0.05).此外,随RIR时间的延长,miR-181a的表达量及RGCs的数量均逐渐减少,两者呈正相关(r=0.995,P=0.005).TNF-α主要在内层视网膜表达,与miR-181a分布重叠;RIR即刻至24 h之间TNF-α表达明显升高,与RGCs计数及miR-181a表达的变化趋势相反,但总体相关性分析无统计学意义. 结论 在RIR模型中,miR-181a可能参与RGCs凋亡的调控,TNF-α是其可能的下游靶基因之一,RIR 24 h前是重要的干预时机.深入研究miR-181a及其靶向基因有助于探寻新的神经保护治疗靶点.
揹景 視網膜缺血-再灌註(RIR)損傷是眼科常見病理改變之一,但其髮病機製尚未明確.目的探討大鼠RIR模型中微小RNA-181a(miR-181a)與視網膜神經節細胞(RGCs)凋亡之間的關繫及可能的靶嚮機製.方法 構建68隻SD大鼠RIR模型,按隨機數字錶法隨機分為對照組和造模後即刻組、造模後24 h組及造模後72 h組,每組17隻,根據MiRanda、Targetscan和miRBase 3箇靶基因數據庫的共同預測結果,選取腫瘤壞死因子-α(TNF-α)作為miR-181a的下遊靶基因研究對象,通過免疫熒光標記法Western blot 及實時熒光定量PCR法觀察miR-181a、TNF-α在各組的錶達情況及其與RGCs凋亡的關繫. 結果 RGCs計數在造模後24 h和72 h顯著減少,與對照組比較差異有統計學意義(P＜0.001).各造模組miR-181a錶達量隨造模時間明顯下降,與對照組比較差異有統計學意義(P＜0.05).此外,隨RIR時間的延長,miR-181a的錶達量及RGCs的數量均逐漸減少,兩者呈正相關(r=0.995,P=0.005).TNF-α主要在內層視網膜錶達,與miR-181a分佈重疊;RIR即刻至24 h之間TNF-α錶達明顯升高,與RGCs計數及miR-181a錶達的變化趨勢相反,但總體相關性分析無統計學意義. 結論 在RIR模型中,miR-181a可能參與RGCs凋亡的調控,TNF-α是其可能的下遊靶基因之一,RIR 24 h前是重要的榦預時機.深入研究miR-181a及其靶嚮基因有助于探尋新的神經保護治療靶點.
배경 시망막결혈-재관주(RIR)손상시안과상견병리개변지일,단기발병궤제상미명학.목적탐토대서RIR모형중미소RNA-181a(miR-181a)여시망막신경절세포(RGCs)조망지간적관계급가능적파향궤제.방법 구건68지SD대서RIR모형,안수궤수자표법수궤분위대조조화조모후즉각조、조모후24 h조급조모후72 h조,매조17지,근거MiRanda、Targetscan화miRBase 3개파기인수거고적공동예측결과,선취종류배사인자-α(TNF-α)작위miR-181a적하유파기인연구대상,통과면역형광표기법Western blot 급실시형광정량PCR법관찰miR-181a、TNF-α재각조적표체정황급기여RGCs조망적관계. 결과 RGCs계수재조모후24 h화72 h현저감소,여대조조비교차이유통계학의의(P＜0.001).각조모조miR-181a표체량수조모시간명현하강,여대조조비교차이유통계학의의(P＜0.05).차외,수RIR시간적연장,miR-181a적표체량급RGCs적수량균축점감소,량자정정상관(r=0.995,P=0.005).TNF-α주요재내층시망막표체,여miR-181a분포중첩;RIR즉각지24 h지간TNF-α표체명현승고,여RGCs계수급miR-181a표체적변화추세상반,단총체상관성분석무통계학의의. 결론 재RIR모형중,miR-181a가능삼여RGCs조망적조공,TNF-α시기가능적하유파기인지일,RIR 24 h전시중요적간예시궤.심입연구miR-181a급기파향기인유조우탐심신적신경보호치료파점.
Background Retinal ischemia-reperfusion (RIR) injury is a common pathologic change.Its mechanism has not been identified.Objective This study was to investigate the relationship of microRNA-181a (miR-181a) ,tumor necrosis factor-α (TNF-α) and retinal ganglial cells (RGCs) in RIR injury.Methods RIR models were induced in 68 rats,then the rats were randomly divided into control group and RIR groups,including 0hour group,24-hour group and 72-hour group by random number table.Predicted target gene TNF-α was chosen,according to M iRanda,Targetscan and miRBase databases.Immunofluorescent labeling, Western blot and quantitative real-time PCR were used to identify the expression levels of miR-181a,TNF-α and RGCs.Immunofluorescent labeling of RGCs in retinal flat mounts was analyzed for RGCs counts.Results Compared with the control group, RGCs densitiy was obviously decreased in 24-hour and 72-hour RIR groups (P＜0.001).The expression level of mir-181a significantly decreased with reperfusion time in the RIR groups (P＜0.05).Futhermore, the expression level of miR181a was positively correlated with RGCs numbers (r=0.995 ,P=0.005).TNF-α and miR-181a were mainly located in inner layers of retina.As opposed to the changes in RGCs numbers and miR-181a expression,TNF-α in 24-hour group was obviously higher than that of the 0-hour group, though there was no statistical significance in overall correlation analysis.Conclusions In RIR,miR-181a may be involved in regulating RGCs apoptosis.TNF-α may be a target gene of miR-181 a.Interventions within 24 hours after reperfusion might be critical.Further study of miR181 a may help to explore new molecular targets for neuroprotection treatment.
