中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2008年
6期
431-435
,共5页
邹贺%黎晓新%于文贞%闫征%张同河%张晓光
鄒賀%黎曉新%于文貞%閆徵%張同河%張曉光
추하%려효신%우문정%염정%장동하%장효광
血管内皮生长因子类%视网膜新生血管化%整合素类%信号传导%动物实验
血管內皮生長因子類%視網膜新生血管化%整閤素類%信號傳導%動物實驗
혈관내피생장인자류%시망막신생혈관화%정합소류%신호전도%동물실험
Vascular endothelial growth factors%Retinal neovascularization%Integrins%Signal transduction%Animal experimentation
目的 观察整合连接激酶(ILK)在血管内皮生长因子(VEGF)诱导的视网膜新生血管形成过程中的作用.方法 在体外培养的视网膜脉络膜血管内皮细胞(RF/6A细胞系)中,用LY294002抑制ILK活性或siRNA转染敲减ILK表达后,检测ILK对VEGF诱导的细胞黏附、增生、迁移及内皮细胞管状结构形成的作用;并在动物模型水平观察LY294002抑制ILK活性后对视网膜新生血管形成的影响.结果 正常对照组、VEGF处理组、LY294002抑制组、siRNA转染组细胞黏附实验结果分别为(0.0726±0.01961),(0.1137±0.02631)、(0.0837±0.01503)、(0.0853±0.02454),VEGF处理组与正常对照组比较,差异有统计学意义(t=4.211,P<0.01);LY294002抑制组以及siRNA转染组与VEGF处理组相比,差异有统计学意义(t=3.074和2.91,P<0.01).正常对照组、VEGF处理组、LY294002抑制组细胞增生实验结果分别为(0.4162±0.1392)、(0.6412±0.2420)、(0.4476±0.1834),VEGF处理组较正常对照组比较,差异有统计学意义(t=2.608,P<0.05);LY294002抑制组与VEGF处理组相比,差异也有统计学意义(t=2.244,P<0.05).正常对照组、VEGF处理组、LY294002抑制组细胞迁移实验结果分别为(83.66±30.283)、(248±74.748)、(138.5±38.167),VEGF处理组与正常对照组比较,差异有统计学意义(t=5.436,P<0.01);LY294002抑制组与VEGF处理组相比,差异也具有统计学意义(t=3.682,P<0.01).血管内皮细胞管状结构形成实验显示,ILK活性或表达受到抑制后无明显管状结构形成.动物实验显示腹膜下注射LY294002使视网膜后极部无灌注区面积从(62 798±16 995.62)μm2增加至(84 722.65±10 435.01)μm2,二者比较,差异有统计学意义(t=3.476,P<0.01).结论 应用LY294002抑制ILK活性或siRNA转染敲减ILK表达使细胞黏附率、细胞增牛率及细胞迁移率均显著下降.ILK通过参与调控VEGF诱导的视网膜血管内皮细胞黏附、增生、迁移及内皮细胞管状结构形成过程而对VEGF诱导的视网膜新生血管形成过程发挥着重要作用.
目的 觀察整閤連接激酶(ILK)在血管內皮生長因子(VEGF)誘導的視網膜新生血管形成過程中的作用.方法 在體外培養的視網膜脈絡膜血管內皮細胞(RF/6A細胞繫)中,用LY294002抑製ILK活性或siRNA轉染敲減ILK錶達後,檢測ILK對VEGF誘導的細胞黏附、增生、遷移及內皮細胞管狀結構形成的作用;併在動物模型水平觀察LY294002抑製ILK活性後對視網膜新生血管形成的影響.結果 正常對照組、VEGF處理組、LY294002抑製組、siRNA轉染組細胞黏附實驗結果分彆為(0.0726±0.01961),(0.1137±0.02631)、(0.0837±0.01503)、(0.0853±0.02454),VEGF處理組與正常對照組比較,差異有統計學意義(t=4.211,P<0.01);LY294002抑製組以及siRNA轉染組與VEGF處理組相比,差異有統計學意義(t=3.074和2.91,P<0.01).正常對照組、VEGF處理組、LY294002抑製組細胞增生實驗結果分彆為(0.4162±0.1392)、(0.6412±0.2420)、(0.4476±0.1834),VEGF處理組較正常對照組比較,差異有統計學意義(t=2.608,P<0.05);LY294002抑製組與VEGF處理組相比,差異也有統計學意義(t=2.244,P<0.05).正常對照組、VEGF處理組、LY294002抑製組細胞遷移實驗結果分彆為(83.66±30.283)、(248±74.748)、(138.5±38.167),VEGF處理組與正常對照組比較,差異有統計學意義(t=5.436,P<0.01);LY294002抑製組與VEGF處理組相比,差異也具有統計學意義(t=3.682,P<0.01).血管內皮細胞管狀結構形成實驗顯示,ILK活性或錶達受到抑製後無明顯管狀結構形成.動物實驗顯示腹膜下註射LY294002使視網膜後極部無灌註區麵積從(62 798±16 995.62)μm2增加至(84 722.65±10 435.01)μm2,二者比較,差異有統計學意義(t=3.476,P<0.01).結論 應用LY294002抑製ILK活性或siRNA轉染敲減ILK錶達使細胞黏附率、細胞增牛率及細胞遷移率均顯著下降.ILK通過參與調控VEGF誘導的視網膜血管內皮細胞黏附、增生、遷移及內皮細胞管狀結構形成過程而對VEGF誘導的視網膜新生血管形成過程髮揮著重要作用.
목적 관찰정합련접격매(ILK)재혈관내피생장인자(VEGF)유도적시망막신생혈관형성과정중적작용.방법 재체외배양적시망막맥락막혈관내피세포(RF/6A세포계)중,용LY294002억제ILK활성혹siRNA전염고감ILK표체후,검측ILK대VEGF유도적세포점부、증생、천이급내피세포관상결구형성적작용;병재동물모형수평관찰LY294002억제ILK활성후대시망막신생혈관형성적영향.결과 정상대조조、VEGF처리조、LY294002억제조、siRNA전염조세포점부실험결과분별위(0.0726±0.01961),(0.1137±0.02631)、(0.0837±0.01503)、(0.0853±0.02454),VEGF처리조여정상대조조비교,차이유통계학의의(t=4.211,P<0.01);LY294002억제조이급siRNA전염조여VEGF처리조상비,차이유통계학의의(t=3.074화2.91,P<0.01).정상대조조、VEGF처리조、LY294002억제조세포증생실험결과분별위(0.4162±0.1392)、(0.6412±0.2420)、(0.4476±0.1834),VEGF처리조교정상대조조비교,차이유통계학의의(t=2.608,P<0.05);LY294002억제조여VEGF처리조상비,차이야유통계학의의(t=2.244,P<0.05).정상대조조、VEGF처리조、LY294002억제조세포천이실험결과분별위(83.66±30.283)、(248±74.748)、(138.5±38.167),VEGF처리조여정상대조조비교,차이유통계학의의(t=5.436,P<0.01);LY294002억제조여VEGF처리조상비,차이야구유통계학의의(t=3.682,P<0.01).혈관내피세포관상결구형성실험현시,ILK활성혹표체수도억제후무명현관상결구형성.동물실험현시복막하주사LY294002사시망막후겁부무관주구면적종(62 798±16 995.62)μm2증가지(84 722.65±10 435.01)μm2,이자비교,차이유통계학의의(t=3.476,P<0.01).결론 응용LY294002억제ILK활성혹siRNA전염고감ILK표체사세포점부솔、세포증우솔급세포천이솔균현저하강.ILK통과삼여조공VEGF유도적시망막혈관내피세포점부、증생、천이급내피세포관상결구형성과정이대VEGF유도적시망막신생혈관형성과정발휘착중요작용.
Objective To evaluate the effect of integrin-linked kinase(ILK)in the process ofretinal neovascularization induced by vascular endothelial growth factor(VEGF).Methods The ILKactivities of retinal choriodal endothelial cell line RF/6A were inhibited by LY294002 or siRNAknockdown.VEGF-induced changes of cell adhesion,proliferation,migration and endothelial cell tube-formation were measured then.The in-vivo effects of ILK were also assessed by intraperitoneal injection ofLY294002 into an animal model of RNV.Results The cell adhesion measurements of control group,VEGF group,VEGF+LY294002 group and VEGF+siRNA group were 0.0726±0.019 61,0.1137±0.026 31,0.0837±0.01 5 03 and 0.0853±0.024 54,respectively.The difference was statisticallysignificant between VEGF group and control group(t=4.211,P<0.01),and between(VEGF+LY294002)group or(VEGF+siRNA)group and control group(t=3.074,2.91,P<0.01).The cellproliferation results of control group,VEGF group and VEGF+LY294002 group were 0.4162±0.1392,0.6412±0.2420,0.4476±0.1834,respectively.The difference was statistically significant betweenVEGF group and control group(t=2.608,P<0.05),and between(VEGF+LY294002)group and VEGFgroup(t=2.244,P<0.05).The cell migration results of control group,VEGF group and VEGF+LY294002 group were 83.66±30.283,248±74.748,138.5±38.167,respectively.The difference wasstatistically significant between VEGF group and control group(t=5.436,P<0.01),and between(VEGF+LY294002)group and VEGF group(t=3.682,P<0.01).There was no obvious tube-formationafter ILK activity was inhibited or knocked down.The non-perfusion areas were increased from(62 798±16 995.62)μm2 to(84 722.65±10 435.01)μm2 after intraperitoneal injection of LY294002 into animalmodel of RNV,the difference was statistically significant(t=3.476,P<0.01).Conclusions ILK mayplay an important role in the process of VEGF-induced retinal neovascularization by regulating the cellularadhesion,proliferation,migration and tube-formation,as all those cellular functions were supressedobviously after the ILK activity was inhibited by LY294002 or the ILK expression was knocked down bysiRNA.
