中华眼科杂志
中華眼科雜誌
중화안과잡지
Chinese Journal of Ophthalmology
2012年
4期
344-349
,共6页
视网膜新生血管化%黏着斑激酶2%血管内皮生长因子A%视网膜病,早产儿%疾病模型,动物
視網膜新生血管化%黏著斑激酶2%血管內皮生長因子A%視網膜病,早產兒%疾病模型,動物
시망막신생혈관화%점착반격매2%혈관내피생장인자A%시망막병,조산인%질병모형,동물
Retinal neovascularization%Focal adhesion kinase 2%Vascular endothelial growth factor A%Retinopathy of prematurity%Disease models,animal
目的 探讨高氧诱导小鼠视网膜新生血管模型中富含脯氨酸的酪氨酸激酶2(Pyk2)基因表达水平的变化.方法 实验研究.选取7日龄C57 BL/6J新生小鼠144只,分为高氧组和对照组,每组72只.高氧组小鼠在密闭氧箱内以(75±2)％氧浓度饲养5d后置于室内空气环境中；对照组小鼠一直处于室内空气环境中饲养.两组小鼠于出生后12、14、17 d分别处死8只,进行组织病理学切片检查和视网膜铺片.实时荧光定量PCR检测Pyk2和血管内皮生长因子(VEGF)在视网膜组织中的表达水平.多组间均数比较采用两因素方差分析,组间均数比较采用SNK-q检验；同一时间点两组间数据比较采用t检验.结果 组织病理学切片检查结果可见,高氧组小鼠出生后14 d(15.36±3.69)和17 d(29.63 ±4.69)有大量突破视网膜内界膜的内皮细胞核及血管芽,与对照组(0.97±1.00、0.83 ±0.79)相比差异有统计学意义(t14 d=- 20.629,P14d =0.000;t17d=-33.814,P17d=0.000).视网膜铺片检查结果可见,高氧组小鼠出生后12 d视网膜血管明显收缩、阻塞,视网膜大片区域无灌注,出生后14 d新生血管开始形成,出生后17 d新生血管形成达到高峰.实时荧光定量PCR检测结果可见,与对照组(Pyk2:12 d:1.00 ±0.14,14 d:0.71±0.20,17 d:0.67±0.15;VEGF:12 d:1.00±0.13,14 d:0.88 ±0.14,17 d:0.94 ±0.15)相比,高氧组出生后12 d Pyk2(0.05±0.03)和VEGF(0.10 ±0.06) mRNA表达量均降低(t=15.706,P=0.000;t=15.911,P=0.000),而出生后14 d和17 d Pyk2(1.11 ±0.22、1.68±0.30)和VEGF(2.10±0.41、4.85±0.46)表达量增高(Pyk2:t14d=-3.376,P14d =0.007;t17d=-7.358,P17d =0.000;VEGF:t14d=-6.904,P14d=0.000;f17d=- 19.667,P17d=0.000).结论 高氧诱导小鼠出生后14 d和17 d视网膜组织中Pyk2的表达水平增高,Pvk2可能参与高氧诱导的视网膜新生血管形成.
目的 探討高氧誘導小鼠視網膜新生血管模型中富含脯氨痠的酪氨痠激酶2(Pyk2)基因錶達水平的變化.方法 實驗研究.選取7日齡C57 BL/6J新生小鼠144隻,分為高氧組和對照組,每組72隻.高氧組小鼠在密閉氧箱內以(75±2)％氧濃度飼養5d後置于室內空氣環境中；對照組小鼠一直處于室內空氣環境中飼養.兩組小鼠于齣生後12、14、17 d分彆處死8隻,進行組織病理學切片檢查和視網膜鋪片.實時熒光定量PCR檢測Pyk2和血管內皮生長因子(VEGF)在視網膜組織中的錶達水平.多組間均數比較採用兩因素方差分析,組間均數比較採用SNK-q檢驗；同一時間點兩組間數據比較採用t檢驗.結果 組織病理學切片檢查結果可見,高氧組小鼠齣生後14 d(15.36±3.69)和17 d(29.63 ±4.69)有大量突破視網膜內界膜的內皮細胞覈及血管芽,與對照組(0.97±1.00、0.83 ±0.79)相比差異有統計學意義(t14 d=- 20.629,P14d =0.000;t17d=-33.814,P17d=0.000).視網膜鋪片檢查結果可見,高氧組小鼠齣生後12 d視網膜血管明顯收縮、阻塞,視網膜大片區域無灌註,齣生後14 d新生血管開始形成,齣生後17 d新生血管形成達到高峰.實時熒光定量PCR檢測結果可見,與對照組(Pyk2:12 d:1.00 ±0.14,14 d:0.71±0.20,17 d:0.67±0.15;VEGF:12 d:1.00±0.13,14 d:0.88 ±0.14,17 d:0.94 ±0.15)相比,高氧組齣生後12 d Pyk2(0.05±0.03)和VEGF(0.10 ±0.06) mRNA錶達量均降低(t=15.706,P=0.000;t=15.911,P=0.000),而齣生後14 d和17 d Pyk2(1.11 ±0.22、1.68±0.30)和VEGF(2.10±0.41、4.85±0.46)錶達量增高(Pyk2:t14d=-3.376,P14d =0.007;t17d=-7.358,P17d =0.000;VEGF:t14d=-6.904,P14d=0.000;f17d=- 19.667,P17d=0.000).結論 高氧誘導小鼠齣生後14 d和17 d視網膜組織中Pyk2的錶達水平增高,Pvk2可能參與高氧誘導的視網膜新生血管形成.
목적 탐토고양유도소서시망막신생혈관모형중부함포안산적락안산격매2(Pyk2)기인표체수평적변화.방법 실험연구.선취7일령C57 BL/6J신생소서144지,분위고양조화대조조,매조72지.고양조소서재밀폐양상내이(75±2)％양농도사양5d후치우실내공기배경중；대조조소서일직처우실내공기배경중사양.량조소서우출생후12、14、17 d분별처사8지,진행조직병이학절편검사화시망막포편.실시형광정량PCR검측Pyk2화혈관내피생장인자(VEGF)재시망막조직중적표체수평.다조간균수비교채용량인소방차분석,조간균수비교채용SNK-q검험；동일시간점량조간수거비교채용t검험.결과 조직병이학절편검사결과가견,고양조소서출생후14 d(15.36±3.69)화17 d(29.63 ±4.69)유대량돌파시망막내계막적내피세포핵급혈관아,여대조조(0.97±1.00、0.83 ±0.79)상비차이유통계학의의(t14 d=- 20.629,P14d =0.000;t17d=-33.814,P17d=0.000).시망막포편검사결과가견,고양조소서출생후12 d시망막혈관명현수축、조새,시망막대편구역무관주,출생후14 d신생혈관개시형성,출생후17 d신생혈관형성체도고봉.실시형광정량PCR검측결과가견,여대조조(Pyk2:12 d:1.00 ±0.14,14 d:0.71±0.20,17 d:0.67±0.15;VEGF:12 d:1.00±0.13,14 d:0.88 ±0.14,17 d:0.94 ±0.15)상비,고양조출생후12 d Pyk2(0.05±0.03)화VEGF(0.10 ±0.06) mRNA표체량균강저(t=15.706,P=0.000;t=15.911,P=0.000),이출생후14 d화17 d Pyk2(1.11 ±0.22、1.68±0.30)화VEGF(2.10±0.41、4.85±0.46)표체량증고(Pyk2:t14d=-3.376,P14d =0.007;t17d=-7.358,P17d =0.000;VEGF:t14d=-6.904,P14d=0.000;f17d=- 19.667,P17d=0.000).결론 고양유도소서출생후14 d화17 d시망막조직중Pyk2적표체수평증고,Pvk2가능삼여고양유도적시망막신생혈관형성.
Objective To analyze the variation of expression of proline-rich tyrosine kinase 2 (Pyk2) in the oxygen-induced retinal neovascularization mice model.Methods Experimental study.One hundred and forty-four C57BL/6J mice were divided equally into the hyperoxia group and the control group.In the hyperoxia group,72 mice (7-day-old) were exposed to (75 ± 2)％ oxygen for 5 days and then moved to room air; in the control group,72 mice were simply raised in room air.These mice were sacrificed on the 12th,14th,17th days and their eyeballs were collected for the preparation of pathological section,retina fiat mounting and RNA extraction.The expression of Pyk2 and vascular endothelial growth factor (VEGF)mRNA in the retina were measured by real-time PCR.ANOVA was used in conjunction with SNK-q test to assess statistical significance at different time within groups ; t-test was used to assess statistical significance between two groups at the same time point.Results Pathological sections showed that there were many endothelial cell nucleus and vascular buds on the 14th day (15.36 ±3.69) and 17th day (29.63 ±4.69) in hyperoxia group.There was significant difference between the control group (0.97 ± 1.00,0.83 ± 0.79 ) and hyperoxia group ( t14 d =- 20.629,P14 d =0.000 ; t17d =- 33.814,P17 d =0.000 ).Retina flat mounting showed that on the 12th day,hyperoxia group showed vascular occlusion,vasoconstriction and large nonperfusion areas,neovascularization appeared and reached the peak on the 17th day.Real-time PCR showed that on the 12th day in hyperoxia group,Pyk2 mRNA (0.05 ±0.03) and VEGF mRNA (0.10 ±0.06) were lower ( Pyk2:t =15.706,P =0.000 ; VEGF:t =15.911,P =0.000 ).However,on the 14 th day and the 17th day,Pyk2 mRNA ( 1.11 ± 0.22,1.68 ± 0.30) and VEGF mRNA (2.10 ± 0.41,4.85 ± 0.46) increased significantly ( Pyk2:t14 d =- 3.376,P14 d =0.007 ; t17 d =- 7.358,P17 d =0.000 ; VEGF,t14 d =- 6.904,P14d =0.000;t17d =- 19.667,P17d =0.000).Conclusion Comparing to the control group,expression of Pyk2 in retinal tissue increases on the 14th day and 17th day in the hyperoxia group,indicating that the expression of Pvk2 is correlated with neovascularization.
