CAJ | 학술논문

目的观察腺苷A2A受体在小鼠视网膜病理性血管形成中的作用.方法将202只新生小鼠分为空气组和氧诱导视网膜病变(OIR)组,分别为66、136只.空气组再分为A2A基因敲除空气组、野生型空气组、咖啡因饮水空气组,分别为18、24、24只,均置于正常空气浓度中饲养.OIR组再分为野生型氧诱导组、A2A基因敲除氧诱导组及咖啡因饮水氧诱导组,分别为48、24、64只,建立小鼠OIR模型.行视网膜石蜡切片,观察病理性新生血管反应;荧光定量聚合酶链反应(PCR)检测小鼠视网膜组织中A2A和血管内皮生长因子(VEGF)的mRNA表达.将0.1、0.3、1.0 g/L剂量的腺苷受体抑制剂咖啡因溶于咖啡因饮水氧诱导组哺乳母鼠饮用水中,定量分析不同剂量及当剂量为1.0 g/L时,出生后0～17、0～7、7～17、7～12、12～17 d等不同饮水时间窗的小鼠视网膜无血管区面积比.结果与野生型氧诱导组相比,A2A基因敲除氧诱导组视网膜中周部无血管区面积明显减少,差异有统计学意义(t=7.694,P＜0.001);突破内界膜的细胞数也明显减少,差异有统计学意义(t=7.747,P＜0.001).荧光定量PCR检测结果显示,与野生型空气组比较,野生型氧诱导组小鼠视网膜中A2A、VEGF mRNA表达量均增高,差异有统计学意义(t=4.036、2.230,P＜0.05).与野生型氧诱导组比较,A2A基因敲除氧诱导组小鼠视网膜中VEGFmRNA表达明显下降,差异有统计学意义(t=3.122,P＜0.01).与野生型氧诱导组比较,0.1、1.0 g/L剂量组小鼠视网膜无血管区面积明显减少,差异有统计学意义(t=2.397、4.533,P＜0.05);出生后0～17、0～7 d饮水时间窗的小鼠视网膜无血管区面积明显减少,差异有统计学意义(t=4.070、2.399,P＜0.05).结论腺苷A2A受体在诱导小鼠视网膜病理性血管形成时表达升高.腺苷A2A受体可调节VEGF的表达.A2A受体失活特异性抑制病理性视网膜新生血管.
목적 관찰선감A2A수체재소서시망막병이성혈관형성중적작용.방법 장202지신생소서분위공기조화양유도시망막병변(OIR)조,분별위66、136지.공기조재분위A2A기인고제공기조、야생형공기조、가배인음수공기조,분별위18、24、24지,균치우정상공기농도중사양.OIR조재분위야생형양유도조、A2A기인고제양유도조급가배인음수양유도조,분별위48、24、64지,건립소서OIR모형.행시망막석사절편,관찰병이성신생혈관반응;형광정량취합매련반응(PCR)검측소서시망막조직중A2A화혈관내피생장인자(VEGF)적mRNA표체.장0.1、0.3、1.0 g/L제량적선감수체억제제가배인용우가배인음수양유도조포유모서음용수중,정량분석불동제량급당제량위1.0 g/L시,출생후0～17、0～7、7～17、7～12、12～17 d등불동음수시간창적소서시망막무혈관구면적비.결과 여야생형양유도조상비,A2A기인고제양유도조시망막중주부무혈관구면적명현감소,차이유통계학의의(t=7.694,P＜0.001);돌파내계막적세포수야명현감소,차이유통계학의의(t=7.747,P＜0.001).형광정량PCR검측결과현시,여야생형공기조비교,야생형양유도조소서시망막중A2A、VEGF mRNA표체량균증고,차이유통계학의의(t=4.036、2.230,P＜0.05).여야생형양유도조비교,A2A기인고제양유도조소서시망막중VEGFmRNA표체명현하강,차이유통계학의의(t=3.122,P＜0.01).여야생형양유도조비교,0.1、1.0 g/L제량조소서시망막무혈관구면적명현감소,차이유통계학의의(t=2.397、4.533,P＜0.05);출생후0～17、0～7 d음수시간창적소서시망막무혈관구면적명현감소,차이유통계학의의(t=4.070、2.399,P＜0.05).결론 선감A2A수체재유도소서시망막병이성혈관형성시표체승고.선감A2A수체가조절VEGF적표체.A2A수체실활특이성억제병이성시망막신생혈관.
Objective To investigate the role of adenosine A2A receptor plays in retinal pathological neovascularization in mice.Methods A total of 202 mice were divided into room-air group (n =66) and oxygen induced retinopathy (OIR) group (n=136).Among room-air group,there were 18 A2A knock-out (KO) mice (KO subgroup) and 24 C57BL/6 mice as wide type (wide type subgroup).OIR group were divided into OIR control subgroup (n =48),A2A-OIR subgroup (n =24) and Caffeine-OIR subgroup (n =64).The retinal neovascularization of OIR group was induced by oxygen.The pathological neovascularization was determined by retinal sections.Fluorescent quantitative polymerase chain reaction (PCR) was used to measure the mRNA expression of A2A and vascular endothelial growth factor (VEGF).0.1,0.3,1.0 g/L Caffeine was dissolve in drinking water of lactating females in Caffeine-OIR subgroup,non-perfusion areas of retina in mice at the age of 0-17,0-7,7-17,7-12,and 12-17 days were analyzed in different dosage and when the dosage as 1.0 g/L.Results Compared with OIR control subgroup,the retinal non-perfusion areas and the numbers of endothelial cell nuclei breaking through the internal limiting membrane in A2A-OIR subgroup were reduced significantly (t =7.694,7.747; P ＜0.001).Compared with wide type subgroup,the level of A2A and VEGF mRNA in OIR control subgroup increased significantly (t=4.036,2.230; P＜0.05).Compared with OIR control subgroup,the level of VEGF mRNA in A2A-OIR subgroup decreased significantly (t=3.122,P＜0.01).Compared with OIR control subgroup,the retinal non perfusion areas in mice at the dosage of 0.1 and 1.0 g/L (t=2.397,4.533) and at the age of 0-17,0-7 days when the dosage as 1.0 g/L (t=4.070,2.399) were reduced significantly (P＜0.05).Conclusions The expression of adenosine A2A receptor increases in oxygeninduced retinal pathological neovascularization..Adenosine A2A receptor may regulate the expression of VEGF.A2A receptor inactivation can inhibit oxygen-induced retinal pathological neovascularization.