重组干扰素诱导蛋白-10对视网膜血管内皮细胞增生、迁移及管腔形成的影响
Effects of interferon-inducible protein-10 on proliferation, migration and capillary tube formation of retinal vascular endothelial cells
摘要目的 观察重组干扰素诱导蛋白-10(IP-10)对人视网膜血管内皮细胞(HREC)和人脐静脉血管内皮细胞(HUVEC)增生、迁移及管腔形成能力的影响.方法 逆转录聚合酶链反应(RT-PCR)法检测HREC和HUVEC中CXC趋化因子受体3(CXCR3) mRNA的表达情况;以细胞计数试剂盒-8(CCK-8)增生分析法比较HREC和HUVEC在不同IP-10浓度下增生能力的差异;采用细胞划痕法测定IP-10对HREC和HUVEC迁移的作用;采用基质体外三维成型法检测IP-10对HREC和HUVEC管腔形成的影响.结果 HREC和HUVEC均在基因水平表达CXCR3.CCK-8增生分析法检测结果显示IP-10抑制HREC增生,并呈剂量依赖性(F=6.202,P<0.05),但IP-10对HUVEC增生无明显影响(F=1.183,P>0.05).细胞划痕法测定结果显示,HREC和HUVEC在10、100 ng/mlIP-10干预时的迁移距离均小于对照组迁移距离,差异有统计学意义(F=25.373、23.858,P<0.05).10、100 ng/ml IP-10对HREC完整管腔形成数量无明显影响,1000 ng/ml IP-10可使HREC完整管腔形成数量明显减少;10、100、1000 ng/mlIP-10干预和未行IP-10干预的HREC完整管腔形成数量比较,差异有统计学意义(F=5.359,P<0.05).结论 IP-10对HREC的增生、迁移、管腔形成均有抑制作用,对HUVEC的迁移有抑制作用.
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abstractsObjective The observe the effects of interferon-inducible protein-10 (IP-10) on proliferation,migration and capillary tube formation of human retinal vascular endothelial cells (HREC) and human umbilical vein endothelial cells (HUVEC).Methods The chemokine receptor (CXCR3) mRNA of HREC and HUVEC were quantified by reverse transcriptase polymerase chain reaction (RT-PCR).In the presence of the different concentrations of IP-10,the difference in proliferation capacity of HREC and HUVEC were analyzed by cell counting kit-8 (CCK-8) methods.Wound scratch assay and three-dimensional in vitro matrigel assay were used for measuring migration and capillary tube formation of HREC and HUVEC,respectively.Results RT-PCR revealed both HREC and HUVEC expressed CXCR3.The proliferation of HREC in the presence of IP-10 was inhibited in a dosage-dependent manner (F=6.202,P<0.05),while IP-10 showed no effect on the inhibitory rate of proliferation of HUVEC (F=1.183,P>0.05).Wound scratch assay showed a significant reduction in the migrated distance of HREC and HUVEC under 10 ng/ml or 100 ng/ml IP-10 stimulation (F=25.373,23.858; P<0.05).There was no effect on the number of intact tubules formed by HREC in the presence of 10 ng/ml or 100 ng/ml IP-10.The number of intact tubules formed by HREC in the presence of 1000 ng/ml IP-10 was remarkably smaller.The difference of number of intact tubules formed by HREC among 10,100,1000 ng/ml 1P-10 and nonintervention group was statistically significant (F=5.359,P<0.05).Conclusion IP-10 can inhibit the proliferation,migration and capillary tube formation ability of HREC and the migration of HUVEC.
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