中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2015年
1期
72-76
,共5页
视网膜新生血管化/药物疗法%富含半胱氨酸蛋白质61%RNA干扰%动物实验
視網膜新生血管化/藥物療法%富含半胱氨痠蛋白質61%RNA榦擾%動物實驗
시망막신생혈관화/약물요법%부함반광안산단백질61%RNA간우%동물실험
Retinal neovascularization/drug therapy%Cysteine-rich protein 61%RNA interference%Animal experimentation
目的 观察特异性抑制富含半胱氨酸蛋白61(CCN1)对氧诱导视网膜病变(OIR)模型小鼠视网膜新生血管(RNV)的抑制作用.方法 7日龄C57BL/6J小鼠120只,随机分为对照组和实验组,每组各60只.参照文献方法制备OIR模型.小鼠出氧箱前1d即鼠龄11d时,对照组、实验组小鼠玻璃体腔分别注射空载体质粒、CCN1siRNA表达质粒各1μl.视网膜铺片观察视网膜血管形态,病理切片计数突破视网膜内界膜的血管内皮细胞核数,免疫组织化学、蛋白免疫印迹法、实时定量聚合酶链反应检测CCN1、血管内皮生长因子(VEGF)蛋白及mRNA的表达情况.结果 与对照组比较,实验组小鼠视网膜血管分布规则、分支良好、新生血管密度减少.两组间突破视网膜内界膜的血管内皮细胞核数量比较,差异有统计学意义(t=8.756,P＜0.05);CCN1、VEGF蛋白表达量比较,差异均有统计学意义(t=3.253、5.365,P＜0.05);CCN1、VEGF蛋白相对表达量比较,差异均有统计学意义(t=4.573、5.323,P＜0.05);CCN1、VEGFmRNA相对表达量比较,差异均有统计学意义(t=6.724、9.153,P＜0.05).结论 特异性抑制CCN1能有效抑制OIR模型小鼠RNV形成.
目的 觀察特異性抑製富含半胱氨痠蛋白61(CCN1)對氧誘導視網膜病變(OIR)模型小鼠視網膜新生血管(RNV)的抑製作用.方法 7日齡C57BL/6J小鼠120隻,隨機分為對照組和實驗組,每組各60隻.參照文獻方法製備OIR模型.小鼠齣氧箱前1d即鼠齡11d時,對照組、實驗組小鼠玻璃體腔分彆註射空載體質粒、CCN1siRNA錶達質粒各1μl.視網膜鋪片觀察視網膜血管形態,病理切片計數突破視網膜內界膜的血管內皮細胞覈數,免疫組織化學、蛋白免疫印跡法、實時定量聚閤酶鏈反應檢測CCN1、血管內皮生長因子(VEGF)蛋白及mRNA的錶達情況.結果 與對照組比較,實驗組小鼠視網膜血管分佈規則、分支良好、新生血管密度減少.兩組間突破視網膜內界膜的血管內皮細胞覈數量比較,差異有統計學意義(t=8.756,P＜0.05);CCN1、VEGF蛋白錶達量比較,差異均有統計學意義(t=3.253、5.365,P＜0.05);CCN1、VEGF蛋白相對錶達量比較,差異均有統計學意義(t=4.573、5.323,P＜0.05);CCN1、VEGFmRNA相對錶達量比較,差異均有統計學意義(t=6.724、9.153,P＜0.05).結論 特異性抑製CCN1能有效抑製OIR模型小鼠RNV形成.
목적 관찰특이성억제부함반광안산단백61(CCN1)대양유도시망막병변(OIR)모형소서시망막신생혈관(RNV)적억제작용.방법 7일령C57BL/6J소서120지,수궤분위대조조화실험조,매조각60지.삼조문헌방법제비OIR모형.소서출양상전1d즉서령11d시,대조조、실험조소서파리체강분별주사공재체질립、CCN1siRNA표체질립각1μl.시망막포편관찰시망막혈관형태,병리절편계수돌파시망막내계막적혈관내피세포핵수,면역조직화학、단백면역인적법、실시정량취합매련반응검측CCN1、혈관내피생장인자(VEGF)단백급mRNA적표체정황.결과 여대조조비교,실험조소서시망막혈관분포규칙、분지량호、신생혈관밀도감소.량조간돌파시망막내계막적혈관내피세포핵수량비교,차이유통계학의의(t=8.756,P＜0.05);CCN1、VEGF단백표체량비교,차이균유통계학의의(t=3.253、5.365,P＜0.05);CCN1、VEGF단백상대표체량비교,차이균유통계학의의(t=4.573、5.323,P＜0.05);CCN1、VEGFmRNA상대표체량비교,차이균유통계학의의(t=6.724、9.153,P＜0.05).결론 특이성억제CCN1능유효억제OIR모형소서RNV형성.
Objective To explore the inhibition effect of Cysteine-rich 61 (CCN1; Cyr61) specific siRNA expression vector on RNV in a mouse model of oxygen-induced retinopathy (OIR).Methods One hundred and twenty healthy C57BL/6J mice were chosen and randomly divided into the experimental group and control group,with 60 mice in each group.The experimental group was intravitreously injected with CCN1siRNA recombinant plasmids.The control group was injected with vector plasmids.Adenosine diphosphate-ase stained retina flat-mounts was performed to assess the retinal vascular profiles,retinal section with HE staining was applied to count the number of new vascular cell nuclei and the protein and mRNA expression of CCN1 and vascular endothelial growth factor (VEGF) were detected by immunohistochemistry,Western blot and Real-time RT-PCR.Results Compared with control group,regular distributions,good branches and reduced density of retinal neovascularization were observed in the experimental group.The number of nucleus of vascular endothelial cells breaking through the inner limiting membrane was obviously less in the experimental group than that in the control group (t=8.756,P＜ 0.05).The expression of CCN1 and VEGF were obviously decreased in the experimental group compared with the control group (all P＜0.05).Conclusion The development of RNV of ROP can be markedly inhibited by RNA interference targeting CCN1,and CCNlsiRNA may provide an effective method for preventing vascular proliferative retinopathy.
