中华眼科杂志
中華眼科雜誌
중화안과잡지
Chinese Journal of Ophthalmology
2012年
4期
350-355
,共6页
徐庆玥%袁源智%王历阳%袁非
徐慶玥%袁源智%王歷暘%袁非
서경모%원원지%왕력양%원비
视网膜新生血管化%趋化因子CXCL12%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%趨化因子CXCL12%血管內皮生長因子A%視網膜病,早產兒%疾病模型,動物
시망막신생혈관화%추화인자CXCL12%혈관내피생장인자A%시망막병,조산인%질병모형,동물
Retinal neovascularization%Chemokine CXCL12%Vascular endothelial growth factor A%Retinopathy of prematurity%Disease models,animals
目的 探讨基质细胞衍生因子-1(SDF-1)的受体拮抗剂AMD3100对高氧诱导的小鼠视网膜病变(OIR)模型中新生血管的抑制作用.方法 实验研究.58只7日龄C57BL/6小鼠随机分为单纯对照组(n=17)、单纯给氧组(n=17)和给药组(n=24).根据AMD3100的浓度,将给药组再分为小剂量抑制组、大剂量抑制组、小剂量安全对照组、大剂量安全对照组(每组6只).各组小鼠19日龄时行视网膜铺片ADP酶染色、石蜡切片HE染色、抗血管内皮生长因子(VEGF)及抗SDF-1免疫组织化学染色,显微镜下观察视网膜平均阳性染色面积百分比.实时荧光定量PCR法检测视网膜中SDF-1 mRNA及VEGF mRNA表达水平.采用t检验进行组间数据的比较.结果 实时荧光定量PCR法检测结果发现,单纯对照组和单纯给氧组视网膜上均有VEGF mRNA(0.080±0.022和0.123 ±0.032)和SDF-1 mRNA(0.731 ±0.099和0.544±0.108)的表达,且给氧组的表达水平显著增高(t=2.488,P:0.038；t=2.864,P =0.021).石蜡切片检测结果显示,大、小剂量抑制组与其自身对照组相比差异均有统计学意义(t=-9.507,P=0.000;t=-10.761,P=0.000),且视网膜铺片ADP酶染色的结果更接近于正常对照组.各组视网膜的免疫组织化学切片均可见神经上皮有VEGF和SDF-1蛋白表达,但给药组的表达水平比自身对照组低(VEGF:t-7.249,P =0.000;t=-9.02,P =0.000;SDF-1:t=-5.246,P =0.000;t=-5.216,P=0.000).结论 AMD3100可能通过拮抗SDF-1的受体来减少SDF-1与CXCR4相结合所产生的效应,从而使VEGF的蛋白表达水平降低,抑制新生血管生成.
目的 探討基質細胞衍生因子-1(SDF-1)的受體拮抗劑AMD3100對高氧誘導的小鼠視網膜病變(OIR)模型中新生血管的抑製作用.方法 實驗研究.58隻7日齡C57BL/6小鼠隨機分為單純對照組(n=17)、單純給氧組(n=17)和給藥組(n=24).根據AMD3100的濃度,將給藥組再分為小劑量抑製組、大劑量抑製組、小劑量安全對照組、大劑量安全對照組(每組6隻).各組小鼠19日齡時行視網膜鋪片ADP酶染色、石蠟切片HE染色、抗血管內皮生長因子(VEGF)及抗SDF-1免疫組織化學染色,顯微鏡下觀察視網膜平均暘性染色麵積百分比.實時熒光定量PCR法檢測視網膜中SDF-1 mRNA及VEGF mRNA錶達水平.採用t檢驗進行組間數據的比較.結果 實時熒光定量PCR法檢測結果髮現,單純對照組和單純給氧組視網膜上均有VEGF mRNA(0.080±0.022和0.123 ±0.032)和SDF-1 mRNA(0.731 ±0.099和0.544±0.108)的錶達,且給氧組的錶達水平顯著增高(t=2.488,P:0.038；t=2.864,P =0.021).石蠟切片檢測結果顯示,大、小劑量抑製組與其自身對照組相比差異均有統計學意義(t=-9.507,P=0.000;t=-10.761,P=0.000),且視網膜鋪片ADP酶染色的結果更接近于正常對照組.各組視網膜的免疫組織化學切片均可見神經上皮有VEGF和SDF-1蛋白錶達,但給藥組的錶達水平比自身對照組低(VEGF:t-7.249,P =0.000;t=-9.02,P =0.000;SDF-1:t=-5.246,P =0.000;t=-5.216,P=0.000).結論 AMD3100可能通過拮抗SDF-1的受體來減少SDF-1與CXCR4相結閤所產生的效應,從而使VEGF的蛋白錶達水平降低,抑製新生血管生成.
목적 탐토기질세포연생인자-1(SDF-1)적수체길항제AMD3100대고양유도적소서시망막병변(OIR)모형중신생혈관적억제작용.방법 실험연구.58지7일령C57BL/6소서수궤분위단순대조조(n=17)、단순급양조(n=17)화급약조(n=24).근거AMD3100적농도,장급약조재분위소제량억제조、대제량억제조、소제량안전대조조、대제량안전대조조(매조6지).각조소서19일령시행시망막포편ADP매염색、석사절편HE염색、항혈관내피생장인자(VEGF)급항SDF-1면역조직화학염색,현미경하관찰시망막평균양성염색면적백분비.실시형광정량PCR법검측시망막중SDF-1 mRNA급VEGF mRNA표체수평.채용t검험진행조간수거적비교.결과 실시형광정량PCR법검측결과발현,단순대조조화단순급양조시망막상균유VEGF mRNA(0.080±0.022화0.123 ±0.032)화SDF-1 mRNA(0.731 ±0.099화0.544±0.108)적표체,차급양조적표체수평현저증고(t=2.488,P:0.038；t=2.864,P =0.021).석사절편검측결과현시,대、소제량억제조여기자신대조조상비차이균유통계학의의(t=-9.507,P=0.000;t=-10.761,P=0.000),차시망막포편ADP매염색적결과경접근우정상대조조.각조시망막적면역조직화학절편균가견신경상피유VEGF화SDF-1단백표체,단급약조적표체수평비자신대조조저(VEGF:t-7.249,P =0.000;t=-9.02,P =0.000;SDF-1:t=-5.246,P =0.000;t=-5.216,P=0.000).결론 AMD3100가능통과길항SDF-1적수체래감소SDF-1여CXCR4상결합소산생적효응,종이사VEGF적단백표체수평강저,억제신생혈관생성.
Objective To observe the inhibition of neovascularation in the oxygen-induced retinopathy (OIR) mice by a stromal cell-derived factor 1 (SDF-1) antagonist.Methods Experimental study.Fifty-eight 7-day-old C57BL/6 mice were divided into 3 groups randomly,the control group (n =17),the test group (n =17) and the medication group(n =24).According to the dosage of AMD3100,the medication group (n =24) were divided into low dose group,high dose group,low dose control group,and high dose control group (each group n =6).Each group (19-day-old) was sacrificed to perform ADPase staining,paraffin sections and immunohistochemical staining ( anti-VEGF and anti-SDF-1 ). The average positive staining area percentage (APSAP) was measured as the outcomes and processed with the Students' t-test. Results Real-time PCR showed expression of both VEGF mRNA (0.080 ± 0.022 vs.0.123 ±0.032) and SDF-1 mRNA (0.731 ±0.099 vs.0.544 ±0.108) in retinas from the control group and test group,respectively.The expression of these factors in the test group was significantly higher (t =2.488,P =0.038 ; t =2.864,P =0.021 ).The number of neovascular endothelial nuclear that broke through the retinal internal limiting membrane in the paraffin section in the high dose group and the low dose group was significantly less than that in the self-control group ( t =- 9.507,P =0.000 ; t =- 10.761,P =0.000).The appearance of ADPase staining sections in the medication group was more similar to the simple control group than that of the test group. Immumohistochemical staining sections showed that VEGF and SDF-1 expressed in neuroepithelial cells in each group. APSAP in the high dose group and the low dose group was significantly lower than that in the self-control group ( VEGF:t =- 7.249,P =0.000 ; t =-9.02,P=0.000; SDF-1:t=-5.246,P=0.000; t=-5.216,P=0.000).Conclusion These results indicate that AMD3100 block the SDF-1 receptor to reduce the effect of SDF-1,decrease the production of VEGF protein and inhibite neovascularization.
