中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2014年
4期
390-394
,共5页
贺涛%江黎%程谷萌%邢怡桥
賀濤%江黎%程穀萌%邢怡橋
하도%강려%정곡맹%형이교
视网膜新生血管化/预防和控制%花生四烯酸盐5脂氧合酶%细胞外信号调节MAP激酶类%动物实验
視網膜新生血管化/預防和控製%花生四烯痠鹽5脂氧閤酶%細胞外信號調節MAP激酶類%動物實驗
시망막신생혈관화/예방화공제%화생사희산염5지양합매%세포외신호조절MAP격매류%동물실험
Retinal neovascularization/prevention & control%Arachidonate 5-lipoxygenase%Extracellular signal-regulated MAP kinases%Animal experimentation
目的 观察5-脂氧合酶(5-LOX)对缺氧诱导的视网膜新生血管形成的抑制作用,探讨视网膜新生血管形成的可能机制.方法 鼠龄为7d的C57BL/6J幼鼠随机分为正常组、氧诱导视网膜病变(OIR)模型组、大剂量干预组、小剂量干预组、干预对照组.各组分别为12只.除正常组外,其余各组小鼠与哺乳母鼠共同置于氧浓度为(75±2)％的氧箱内饲养5d后转移至正常环境中饲养5d,建立OIR模型.12～17日龄时,大剂量干预组、小剂量干预组小鼠腹腔每天分别注射100、50 mg/kg 5-LOX抑制剂去甲二氢愈创木酸;干预对照组小鼠腹腔每天注射等体积1％二甲基亚砜;OIR模型组小鼠出氧箱后不作任何处理.石蜡切片苏木精-伊红染色并计数突破视网膜内界膜的血管内皮细胞核,CD34标记鼠新生血管内皮细胞,逆转录-聚合酶链反应(RT PCR)检测视网膜中5-LOX、血管内皮生长因子(VEGF)-a、VEGF受体-2(VEGFR-2)的mRNA含量.蛋白质免疫印迹法(Western blot)检测视网膜中5-LOX、VEGF-a、VEGFR-2、细胞外磷酸化信号调节蛋白激酶(P-ERK) 1/2的蛋白表达含量.结果 不同剂量干预组突破内界膜的视网膜新生血管内皮细胞核数较OIR模型组、干预对照组明显减少,差异有统计学意义(F=73.390,P＜0.05).RT-PCR、Western blot检测结果显示,不同剂量干预组小鼠视网膜5-LOX、VEGF-a、VEGFR-2的mRNA和蛋白表达均较OIR模型组、干预对照组减少,差异有统计学意义(F=92.668,P＜0.05);大剂量干预、小剂量干预组之间VEGFR-2蛋白相对表达量比较,差异无统计学意义(F=2.118,P＞0.05);5-LOX、VEGF-a、P-ERK 1/2相对蛋白表达比较,差异均有统计学意义(F=86.490、165.128、139.424,P＜0.05).结论 缺氧可以诱导视网膜中5-LOX的高表达;5 LOX选择性抑制可以降低其表达并减少缺氧诱导的新生血管形成.
目的 觀察5-脂氧閤酶(5-LOX)對缺氧誘導的視網膜新生血管形成的抑製作用,探討視網膜新生血管形成的可能機製.方法 鼠齡為7d的C57BL/6J幼鼠隨機分為正常組、氧誘導視網膜病變(OIR)模型組、大劑量榦預組、小劑量榦預組、榦預對照組.各組分彆為12隻.除正常組外,其餘各組小鼠與哺乳母鼠共同置于氧濃度為(75±2)％的氧箱內飼養5d後轉移至正常環境中飼養5d,建立OIR模型.12～17日齡時,大劑量榦預組、小劑量榦預組小鼠腹腔每天分彆註射100、50 mg/kg 5-LOX抑製劑去甲二氫愈創木痠;榦預對照組小鼠腹腔每天註射等體積1％二甲基亞砜;OIR模型組小鼠齣氧箱後不作任何處理.石蠟切片囌木精-伊紅染色併計數突破視網膜內界膜的血管內皮細胞覈,CD34標記鼠新生血管內皮細胞,逆轉錄-聚閤酶鏈反應(RT PCR)檢測視網膜中5-LOX、血管內皮生長因子(VEGF)-a、VEGF受體-2(VEGFR-2)的mRNA含量.蛋白質免疫印跡法(Western blot)檢測視網膜中5-LOX、VEGF-a、VEGFR-2、細胞外燐痠化信號調節蛋白激酶(P-ERK) 1/2的蛋白錶達含量.結果 不同劑量榦預組突破內界膜的視網膜新生血管內皮細胞覈數較OIR模型組、榦預對照組明顯減少,差異有統計學意義(F=73.390,P＜0.05).RT-PCR、Western blot檢測結果顯示,不同劑量榦預組小鼠視網膜5-LOX、VEGF-a、VEGFR-2的mRNA和蛋白錶達均較OIR模型組、榦預對照組減少,差異有統計學意義(F=92.668,P＜0.05);大劑量榦預、小劑量榦預組之間VEGFR-2蛋白相對錶達量比較,差異無統計學意義(F=2.118,P＞0.05);5-LOX、VEGF-a、P-ERK 1/2相對蛋白錶達比較,差異均有統計學意義(F=86.490、165.128、139.424,P＜0.05).結論 缺氧可以誘導視網膜中5-LOX的高錶達;5 LOX選擇性抑製可以降低其錶達併減少缺氧誘導的新生血管形成.
목적 관찰5-지양합매(5-LOX)대결양유도적시망막신생혈관형성적억제작용,탐토시망막신생혈관형성적가능궤제.방법 서령위7d적C57BL/6J유서수궤분위정상조、양유도시망막병변(OIR)모형조、대제량간예조、소제량간예조、간예대조조.각조분별위12지.제정상조외,기여각조소서여포유모서공동치우양농도위(75±2)％적양상내사양5d후전이지정상배경중사양5d,건립OIR모형.12～17일령시,대제량간예조、소제량간예조소서복강매천분별주사100、50 mg/kg 5-LOX억제제거갑이경유창목산;간예대조조소서복강매천주사등체적1％이갑기아풍;OIR모형조소서출양상후불작임하처리.석사절편소목정-이홍염색병계수돌파시망막내계막적혈관내피세포핵,CD34표기서신생혈관내피세포,역전록-취합매련반응(RT PCR)검측시망막중5-LOX、혈관내피생장인자(VEGF)-a、VEGF수체-2(VEGFR-2)적mRNA함량.단백질면역인적법(Western blot)검측시망막중5-LOX、VEGF-a、VEGFR-2、세포외린산화신호조절단백격매(P-ERK) 1/2적단백표체함량.결과 불동제량간예조돌파내계막적시망막신생혈관내피세포핵수교OIR모형조、간예대조조명현감소,차이유통계학의의(F=73.390,P＜0.05).RT-PCR、Western blot검측결과현시,불동제량간예조소서시망막5-LOX、VEGF-a、VEGFR-2적mRNA화단백표체균교OIR모형조、간예대조조감소,차이유통계학의의(F=92.668,P＜0.05);대제량간예、소제량간예조지간VEGFR-2단백상대표체량비교,차이무통계학의의(F=2.118,P＞0.05);5-LOX、VEGF-a、P-ERK 1/2상대단백표체비교,차이균유통계학의의(F=86.490、165.128、139.424,P＜0.05).결론 결양가이유도시망막중5-LOX적고표체;5 LOX선택성억제가이강저기표체병감소결양유도적신생혈관형성.
Objective To investigate the inhibitory effects of 5-lipoxygenase (5-LOX) on oxygeninduced retinal neovascularization in mice and to explore its possible mechanisms.Methods 7-day-old C57BL/6J mice were randomly divided into normal group,oxygen induced retinopathy (OIR) model group,large-dose group,small-dose group and control group with 12 mice in each group.The mice with their mothers were kept in (75±2)％ of oxygen environment for 5 days and then returned to normoxia for 5 days to establish the OIR model except for normal group.From postnatal day 12 to 17,the large-dose group and small-dose group received intravitreous injection of 5-LOX at dose of 100 mg/kg and 50 mg/kg respectively,while the control group received the same volume of 1 ％ dimethyl sulfoxide.The mice in the OIR group received no treatment.The number of endothelium cell nuclei breaking through the inner limiting membrane (ILM) was counted on hematoxylin and eosin-stained retinal section.The mRNA expression of 5 LOX,vascular endothelial growth factor (VEGF) a,VEGF receptor 2 (VEGFR-2) on retinal tissue were detected by reverse transcription polymerase chain reaction (RT-PCR).The protein expression of 5 LOX,VEGF a,VEGFR-2 and phosphorylation extracellular signal-regulated kinase (P-ERK) 1/2 on retinal tissue were detected by Western blot.Results The number of vascular cell nuclei breaking through the ILM in the large-dose group and small dose group decreased significantly compared with the OIR group and control group (F=73.390,P＜0.05).The mRNA expression and protein expression of 5-LOX,VEGFa,VEGFR 2 on retinal tissue were decreased significantly in the large-dose group and small dose group as compared with the OIR group and control group (F =92.668,P＜ 0.05).The difference of VEGFR-2 protein expression between large-dose group and small dose group was not significant (F=2.118,P＞0.05).The differences of 5-LOX,VEGF a,P-ERK 1/2 protein expression between large-dose group and small-dose group were significant (F=86.490,165.128,139.424; P＜0.05).Conclusion Hypoxia may induce 5 LOX expression in the retina.Retinal neovascularization was significantly inhibited by selective inhibition of 5 LOX.
