目的：观察体外高糖诱导环境，对人视网膜色素上皮细胞中内脏脂肪素(Visfatin)的表达影响，以及研究高糖环境下重楼皂苷I(Polyphyllin I)对Visfatin表达情况的影响。

方法：人视网膜色素上皮细胞培养后分3组，正常对照组、高糖组和高糖加Polyphyllin I药物干预组，干预培养12h后进行检测。正常对照组：5.5mmol/L葡萄糖浓度常规培养； 高糖组：将25mmol/L的高糖加入培养基建立模型； 高糖加Polyphyllin I药物干预组：高糖25mmol/L和3μg/L Polyphyllin I药物加入培养基。免疫荧光染色法观察人视网膜色素上皮细胞中Visfatin和血管内皮细胞生长因子(VEGF)表达； 实时荧光定量PCR检测上皮细胞中Visfatin和VEGF mRNA的表达； Western-blot法检测上皮细胞中Visfatin和VEGF蛋白的表达。

结果：免疫荧光检测发现Visfatin和VEGF在正常组视网膜色素上皮细胞中表达呈弱阳性。但在高糖组视网膜色素上皮中可见Visfatin和VEGF呈强阳性表达。药物干预组中Visfatin和VEGF荧光较高糖组明显减弱。RT-PCR显示高糖组Visfatin mRNA表达水平较正常组和干预组明显增高(t=4.24、3.89，均P<0.05)。高糖组VEGF mRNA表达水平较正常组和干预组明显增高(t=3.53、2.57，均P<0.05)。Western-blot结果示Visfatin蛋白水平，高葡萄糖组表达量显著高于正常对照组(t=3.62，P=0.01)，干预组表达低于高糖组(t=3.79，P<0.01)。

结论：高糖环境可刺激视网膜色素上皮细胞中Visfatin的表达增加，Polyphyllin I可抑制高糖环境下视网膜色素上皮细胞中Visfatin的表达，可能为治疗糖尿病视网膜病变提供新的思路。

METHODS: Human retinal pigment epithelial cells were cultured in three groups, normal control group, high glucose group and intervention group of high glucose aggravated PolyphyllinⅠ, testing after 12h of intervention culture. Normal control group:5.5mmol/L glucose concentration routine culture; high glucose group:25mmol/L high glucose was added to the medium to establish the model; high glucose aggravated PolyphyllinⅠdrug intervention group: high glucose 25mmol/L, 3μg/L PolyphyllinⅠdrug was added to the medium. Immunofluorescence staining assays to observe expression of the Visfatin and VEGF in human retinal pigment epithelial cells; real-time PCR assays for relative expression of Visfatin and VEGF mRNA in epithelial cells; and western-blot assays for Visfatin and VEGF proteins in epithelial cells.

RESULTS: Immunofluorescence detection revealed that Visfatin and VEGF were weakly positive in normal retinal pigment epithelial cells. Visfatin and VEGF were strongly positive in high glucose group. Visfatin and VEGF fluorescence in the drug intervention group was significantly weakened in the higher sugar group. RT-PCR showed that the expression level visfatin mRNA high sugar group was significantly higher than that of normal group and intervention group(t=4.24, 3.89, P<0.05). VEGF mRNA expression was significantly higher in high glucose group than in normal group and intervention group(t=3.53, 2.57, P<0.05). Western-blot results showed that the protein expression of visfatin and VEGF in high sugar group was significantly higher than that in control group and intervention group(t=3.62, P=0.01; t=3.79, P<0.01).

CONCLUSION: The high glucose environment can stimulate the increased expression of Visfatin in retinal pigment epithelial cells, Polyphyllin I can inhibit the expression of Visfatin in retinal pigment epithelial cells in high glucose environment, which may provide a new idea for the treatment of diabetic retinopathy.