海南医学
海南醫學
해남의학
Hainan Medical Journal
2015年
19期
2813-2817
,共5页
张婷%马晓昀%张昀%刘林%丁雯芝%邹俊
張婷%馬曉昀%張昀%劉林%丁雯芝%鄒俊
장정%마효윤%장윤%류림%정문지%추준
角膜新生血管%碱烧伤%雷尼单抗%低氧诱导因子-1α%血管内皮生长因子
角膜新生血管%堿燒傷%雷尼單抗%低氧誘導因子-1α%血管內皮生長因子
각막신생혈관%감소상%뢰니단항%저양유도인자-1α%혈관내피생장인자
Corneal neovascularization%Alkali burn%Ranibizumab%Hypoxia-inducible factor-1α(HIF-1α)%Vascular endothelial growth factor (VEGF)
目的 观察雷尼单抗(Ranibizumab)抑制碱烧伤诱导小鼠角膜新生血管(CorNV)的作用.方法 取清洁级BALB/c小鼠36只,将加有NaOH溶液(1 mol/L)的直径2 mm圆形滤纸片轻贴角膜中央烧灼30 s,制备角膜碱烧伤CorNV模型:右眼接受碱烧伤处理,左眼不予处理.后随机将小鼠分为两组,于碱烧伤后2 d、8 d时右眼分别行球结膜下注射0.9%生理盐水(阴性对照组,18只小鼠)、10 mg/ml Ranibizumab (Ranibizumab治疗组,18只小鼠).碱烧伤后4 d、7d、14 d时行眼前节照相大体观察CorNV生长变化并计算CorNV面积;分别于碱烧伤后7 d、14 d时行苏木素-伊红(HE)染色观察角膜组织病理结构及CorNV变化;免疫组织化学(IHC)染色检测角膜HIF-1α、VEGF-A蛋白表达;实时荧光定量聚合酶链式反应(Real-time PCR)检测角膜HIF-1αmRNA、VEGF-A mRNA表达.结果 眼前节照相、HE染色结果显示:碱烧伤后7 d、14 d时Ranibizumab治疗组较阴性对照组角膜新生血管减少,细小稀疏,CorNV面积明显减小,差异均有统计学意义(P<0.05).PCR结果显示:Ranibizumab治疗组中角膜VEGF-A mRNA相对表达于7 d、14 d时均较阴性对照组降低,7 d时HIF-1αmRNA相对表达降低;差异均有统计学意义(P<0.05).免疫组化结果显示:Ranibizumab治疗组VEGF-A蛋白表达于7 d、14 d时较阴性对照组均降低,差异均有统计学意义(P<0.05);HIF-1α表达未明显改变.结论 Lucentis通过下调VEGF对碱烧伤诱导小鼠角膜CorNV具有一定抑制作用.
目的 觀察雷尼單抗(Ranibizumab)抑製堿燒傷誘導小鼠角膜新生血管(CorNV)的作用.方法 取清潔級BALB/c小鼠36隻,將加有NaOH溶液(1 mol/L)的直徑2 mm圓形濾紙片輕貼角膜中央燒灼30 s,製備角膜堿燒傷CorNV模型:右眼接受堿燒傷處理,左眼不予處理.後隨機將小鼠分為兩組,于堿燒傷後2 d、8 d時右眼分彆行毬結膜下註射0.9%生理鹽水(陰性對照組,18隻小鼠)、10 mg/ml Ranibizumab (Ranibizumab治療組,18隻小鼠).堿燒傷後4 d、7d、14 d時行眼前節照相大體觀察CorNV生長變化併計算CorNV麵積;分彆于堿燒傷後7 d、14 d時行囌木素-伊紅(HE)染色觀察角膜組織病理結構及CorNV變化;免疫組織化學(IHC)染色檢測角膜HIF-1α、VEGF-A蛋白錶達;實時熒光定量聚閤酶鏈式反應(Real-time PCR)檢測角膜HIF-1αmRNA、VEGF-A mRNA錶達.結果 眼前節照相、HE染色結果顯示:堿燒傷後7 d、14 d時Ranibizumab治療組較陰性對照組角膜新生血管減少,細小稀疏,CorNV麵積明顯減小,差異均有統計學意義(P<0.05).PCR結果顯示:Ranibizumab治療組中角膜VEGF-A mRNA相對錶達于7 d、14 d時均較陰性對照組降低,7 d時HIF-1αmRNA相對錶達降低;差異均有統計學意義(P<0.05).免疫組化結果顯示:Ranibizumab治療組VEGF-A蛋白錶達于7 d、14 d時較陰性對照組均降低,差異均有統計學意義(P<0.05);HIF-1α錶達未明顯改變.結論 Lucentis通過下調VEGF對堿燒傷誘導小鼠角膜CorNV具有一定抑製作用.
목적 관찰뢰니단항(Ranibizumab)억제감소상유도소서각막신생혈관(CorNV)적작용.방법 취청길급BALB/c소서36지,장가유NaOH용액(1 mol/L)적직경2 mm원형려지편경첩각막중앙소작30 s,제비각막감소상CorNV모형:우안접수감소상처리,좌안불여처리.후수궤장소서분위량조,우감소상후2 d、8 d시우안분별행구결막하주사0.9%생리염수(음성대조조,18지소서)、10 mg/ml Ranibizumab (Ranibizumab치료조,18지소서).감소상후4 d、7d、14 d시행안전절조상대체관찰CorNV생장변화병계산CorNV면적;분별우감소상후7 d、14 d시행소목소-이홍(HE)염색관찰각막조직병리결구급CorNV변화;면역조직화학(IHC)염색검측각막HIF-1α、VEGF-A단백표체;실시형광정량취합매련식반응(Real-time PCR)검측각막HIF-1αmRNA、VEGF-A mRNA표체.결과 안전절조상、HE염색결과현시:감소상후7 d、14 d시Ranibizumab치료조교음성대조조각막신생혈관감소,세소희소,CorNV면적명현감소,차이균유통계학의의(P<0.05).PCR결과현시:Ranibizumab치료조중각막VEGF-A mRNA상대표체우7 d、14 d시균교음성대조조강저,7 d시HIF-1αmRNA상대표체강저;차이균유통계학의의(P<0.05).면역조화결과현시:Ranibizumab치료조VEGF-A단백표체우7 d、14 d시교음성대조조균강저,차이균유통계학의의(P<0.05);HIF-1α표체미명현개변.결론 Lucentis통과하조VEGF대감소상유도소서각막CorNV구유일정억제작용.
Objective To observe the inhibition effect of Ranibizumab on alkali burn induced-corneal neo-vascularization (CorNV) in mice. Methods Thirty-six healthy BALB/c mice received 1 mol/L NaOH for 30 s to establish models of alkali induced-CorNV of the right eyes. All the mice were randomly divided into two groups, negative control group (eighteen cases, the right eyes received subconjuctival injections of 0.9%normal saline), Ra-nibizumab treatment group (eighteen cases, the right eyes received subconjuctival injections of 10 mg/ml Ranibi-zumab). On days 2, 8 after alkali burn, CorNV was observed and photographed on days 4, 7, 14. The areas of CorNV were calculated. On days 7, 14, hematoxylin-eosin (HE) and immunohistochemical (IHC) staining were con-ducted to observe histopathological change and to detect corneal hypoxia-inducible factor-1α(HIF-1α) and vascu-lar endothelial growth factor (VEGF) expression. Quantitative real-time polymerase chain reaction (real-time PCR) was applied to detect the expression of HIF-1αmRNA and VEGF-A mRNA. Results Frontal section photograph and HE staining revealed that on days 7 and 14 after alkali burn, areas of CorNV in Ranibizumab treatment group were significantly lower than that in negative control group (P<0.05). Real-time PCR showed that relative expres-sion levels of VEGF-A mRNA in Ranibizumab treatment group were all significantly lower than negative control group on day 7 and day 14 after alkali burn, and relative expression levels of HIF-1αmRNA were lower than nega-tive control group on day 7 (all P<0.05). IHC staining revealed that on days 7 and 14 after alkali burn, VEGF-A pro-tein expression in Ranibizumab treatment group decreased compared to negative control group (P<0.05), while HIF-1αprotein expression did not show any significant change. Conclusion Ranibizumab has some inhibition effect on mouse CorNV induced by alkali burn through down-regulation of VEGF.