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
62-66
,共5页
潘颖喆%邢怡桥%陈长征%黎智%王慧%李璐%苏钰
潘穎喆%邢怡橋%陳長徵%黎智%王慧%李璐%囌鈺
반영철%형이교%진장정%려지%왕혜%리로%소옥
视网膜色素上皮%血管内皮生长因子A%受体,CCR3
視網膜色素上皮%血管內皮生長因子A%受體,CCR3
시망막색소상피%혈관내피생장인자A%수체,CCR3
Retinal pigment epithelium%Vascular endothelial growth factor A%Receptors,CCR3
目的 观察抑制光损伤人视网膜色素上皮(RPE)细胞血管内皮生长因子A(VEGF-A)的表达对趋化因子受体3(CCR3)配体嗜酸细胞活化趋化因子(eotaxin)表达的影响.方法 体外培养人RPE细胞,取第8～12代生长良好的传代细胞用于实验.将同代细胞随机分为正常对照组、光照损伤组和光照干预组.采用波长400 nm的蓝色节能灯以(600±100) Lux持续光照RPE细胞12h建立光损伤模型,光照干预组采用0.125 mg/ml的VEGF-A受体拮抗剂雷珠单抗处理光损伤的RPE细胞;正常对照组以锡箔纸包裹细胞培养皿避光培养.结束光照24 h时,应用实时聚合酶链反应、酶联免疫吸附测定、蛋白免疫印迹法及免疫组织化学染色法检测3组细胞中VEGF-A、eotaxin-1、eotaxin-2、eotaxin-3及核因子(NF)-κB的mRNA和蛋白表达情况.结果 光照损伤组VEGF-A、eotaxin-1、eotaxin-2、eotaxin-3、NF-κB的mRNA和蛋白表达较正常对照组明显增加,差异均有统计学意义(P＜0.05).光照干预组VEGF-A、eotaxin-1、eotaxin-2、eotaxin-3、NF-κB的mRNA和蛋白表达较光照损伤组明显降低,差异也有统计学意义(P＜0.05).结论 抑制光损伤后人RPE细胞VEGF-A的表达可以下调CCR3配体eotaxin的表达,其机制可能与阻断转录因子NF-κB的激活有关.
目的 觀察抑製光損傷人視網膜色素上皮(RPE)細胞血管內皮生長因子A(VEGF-A)的錶達對趨化因子受體3(CCR3)配體嗜痠細胞活化趨化因子(eotaxin)錶達的影響.方法 體外培養人RPE細胞,取第8～12代生長良好的傳代細胞用于實驗.將同代細胞隨機分為正常對照組、光照損傷組和光照榦預組.採用波長400 nm的藍色節能燈以(600±100) Lux持續光照RPE細胞12h建立光損傷模型,光照榦預組採用0.125 mg/ml的VEGF-A受體拮抗劑雷珠單抗處理光損傷的RPE細胞;正常對照組以錫箔紙包裹細胞培養皿避光培養.結束光照24 h時,應用實時聚閤酶鏈反應、酶聯免疫吸附測定、蛋白免疫印跡法及免疫組織化學染色法檢測3組細胞中VEGF-A、eotaxin-1、eotaxin-2、eotaxin-3及覈因子(NF)-κB的mRNA和蛋白錶達情況.結果 光照損傷組VEGF-A、eotaxin-1、eotaxin-2、eotaxin-3、NF-κB的mRNA和蛋白錶達較正常對照組明顯增加,差異均有統計學意義(P＜0.05).光照榦預組VEGF-A、eotaxin-1、eotaxin-2、eotaxin-3、NF-κB的mRNA和蛋白錶達較光照損傷組明顯降低,差異也有統計學意義(P＜0.05).結論 抑製光損傷後人RPE細胞VEGF-A的錶達可以下調CCR3配體eotaxin的錶達,其機製可能與阻斷轉錄因子NF-κB的激活有關.
목적 관찰억제광손상인시망막색소상피(RPE)세포혈관내피생장인자A(VEGF-A)적표체대추화인자수체3(CCR3)배체기산세포활화추화인자(eotaxin)표체적영향.방법 체외배양인RPE세포,취제8～12대생장량호적전대세포용우실험.장동대세포수궤분위정상대조조、광조손상조화광조간예조.채용파장400 nm적람색절능등이(600±100) Lux지속광조RPE세포12h건립광손상모형,광조간예조채용0.125 mg/ml적VEGF-A수체길항제뢰주단항처리광손상적RPE세포;정상대조조이석박지포과세포배양명피광배양.결속광조24 h시,응용실시취합매련반응、매련면역흡부측정、단백면역인적법급면역조직화학염색법검측3조세포중VEGF-A、eotaxin-1、eotaxin-2、eotaxin-3급핵인자(NF)-κB적mRNA화단백표체정황.결과 광조손상조VEGF-A、eotaxin-1、eotaxin-2、eotaxin-3、NF-κB적mRNA화단백표체교정상대조조명현증가,차이균유통계학의의(P＜0.05).광조간예조VEGF-A、eotaxin-1、eotaxin-2、eotaxin-3、NF-κB적mRNA화단백표체교광조손상조명현강저,차이야유통계학의의(P＜0.05).결론 억제광손상후인RPE세포VEGF-A적표체가이하조CCR3배체eotaxin적표체,기궤제가능여조단전록인자NF-κB적격활유관.
Objective To observe the changes of eotaxin-1(CCL11),eotaxin-2(CCL24)and eotaxin-3 (CCL26)in ranibizumab treated light-injured human retinal pigment epithelium (RPE) cells ARPE-19 and investigate the effects of vascular endothelial growth factor (VEGF) antagonist to the expressions of eotaxins.Methods Cultured human RPE cells (8th-12th generations)were divided into light-injured group,ranibizumab treated group and normal control group.Cells of the three groups were exposed to the blue light at the intensity of(600± 100) Lux for 12 h to establish the light injured model,while cell culture dishes of the normal control group were wrapped with double-layer foil.The cells of ranibizumab treated group were treated with VEGF-A antagonist(ranibizumab)at the final concentration of 0.125 mg/ml for 24 hours directly after the illumination.The mRNA and protein of VEGF-A,eotaxin-1,eotaxin-2,eotaxin-3,NF-κB were determined by Real time-PCR,enzyme-linked immunosorbent assay,Western blot,immunohistochemical staining at 0,3,6,12,24 hours after light damage.Results The mRNA and protein level of VEGF-A,eotaxin-1,eotaxin-2,eotaxin-3,NF-κB in the light-injuried group increased significantly compared to that in normal control group (P＜0.05).After treating with ranibizumab,the expression of eotaxin-1,eotaxin-2,eotaxin-3,NF-κB were significantly suppressed (P ＜ 0.05).Conclusion The suppression of over-expression of VEGF in human RPE may down-regulate the expression of eotaxins,via the suppression of NF-κB.
