方法：恒河猴脉络膜-视网膜内皮细胞(RF/6A)分别在常氧和低氧环境中培养，分为正常对照组、低氧对照组和治疗组。低氧对照组和治疗组分别采用脂质体Lipofectamine^TM2000(LF2000)介导转染空载体质粒和CCR7siRNA表达质粒。CCK8法检测细胞增殖情况； 流式细胞术检测细胞凋亡情况； Western blot和RT-PCR法检测三组RF/6A内CCR7、VEGF蛋白及mRNA的表达情况。

结果：低氧对照组较正常对照组比较及治疗组较正常对照组和低氧对照组比较细胞生长速度明显减慢，增殖能力减弱，细胞凋亡率显著增加(均为P<0.05)； 低氧对照组与正常对照组比较， RF/6A内CCR7、VEGF蛋白及mRNA表达显著增高，均有统计学意义(t_CCR7蛋白=3.38，t_VEGF蛋白=4.75，t _CCR7mRNA=4.27，t_VEGFmRNA=5.34， 均为P<0.05)，且二者表达呈正相关(r_蛋白=0.71， r _mRNA =0.83， 均为P<0.05)。治疗组CCR7和VEGF的蛋白及mRNA表达较正常对照组和低氧对照组明显下降(均为P<0.05)。

结论：缺氧时REC中CCR7可上调VEGF的表达，CCR7-VEGF信号途径在视网膜新生血管(retinal neovascularization，RNV)形成的过程中可能具有潜在功能，CCR7siRNA有望成为防治RNV的一种有效方法。

METHODS:Rhesus choroid-retinal vascular endothelial cells(RF/6A)were cultured under normoxic and hypoxic conditions in vitro and divided into the normoxia group, the hypoxia control group(transiently transfected with the vector plasmid)and the treated group(transiently transfected with the CCR7siRNA recombinant plasmid). Plasmids were transiently transfected in RF/6A cells using Lipofectamine^TM2000(LF2000). RF/6A cells proliferation and apoptosis detected by CCK8 and flow cytometry and the protein and mRNA expression of CCR7 and VEGF were measured by Western blot and RT-PCR.

RESULTS:The cell growth rate became slower and apoptosis rate became increased in the hypoxia group than that in the normoxia group, and in the treated group than that in the normoxia and hypoxia control groups(all P<0.05). Compared with the normoxia group, there were high protein and mRNA expression of CCR7 and VEGF in the hypoxia control group, the differences had statistical significance(t_CCR7protein=3.38, t_VEGFprotein=4.75, t_CCR7mRNA=4.27, t_VEGFmRNA=5.34, all P<0.05). And there was an obvious positive correlation between the expression of CCR7 and VEGF(r_protein=0.71, r_mRNA=0.83, all P<0.05). The protein and mRNA expression of CCR7 and VEGF were obviously decreased in the treated group compared with the normoxia and hypoxia control groups(all P<0.05).

CONCLUSION: CCR7 can upregulate the expression of VEGF in REC under hypoxia. CCR7- VEGF signaling pathway may have potential function in the retinal neovascularization(RNV), and CCR7siRNA may provide an effective method for RNV.