目的:探讨羟基红花黄色素A(HSYA)对高糖培养下恒河猴脉络膜血管内皮细胞(RF/6A)增殖及血管内皮生长因子(VEGF)mRNA表达的影响。方法:取体外培养的RF/6A细胞,分为NG+H0组(含有5.5mmol/L葡萄糖,HSYA的浓度为0mg/L)及HG+H0、HG+H18、HG+H37、HG+H73组(各组中均含有22mmol/L葡萄糖,HSYA的浓度分别为0,18,37,73mg/L)。培养24,48和72h,分别使用噻唑盐比色法(MTT)检测细胞增殖的活性。采用逆转录-聚合酶链式反应(RT-PCR)检测各组中RF/6A细胞中VEGFmRNA的表达。结果:MTT检测结果表明,HSYA作用48h,HG+H37、HG+H73组细胞吸光度值均明显低于HG+H0组(P<0.01),随着HSYA浓度的增加,各组吸光度值降低。作用72h,HG+H18、HG+H37和HG+H73组细胞吸光度值均明显低于HG+H0组(P<0.01),随着HSYA浓度的增加,各组吸光度值降低,HG+H73组抑制作用最明显,其细胞吸光度值明显低于HG+H18和HG+H37组(分别为P<0.01和P<0.05)。RT-PCR实验结果表明:与HG+H0组相比,作用48h,HG+H37、HG+H73对RF/6A细胞VEGFmRNA表达的抑制作用显著(P<0.01);而作用72h,HG+H18、HG+H37和HG+H73对高糖诱导的VEGFmRNA的表达均有抑制作用(P<0.01)。结论:HSYA能够抑制高糖诱导的RF/6A细胞的增殖并下调高糖所致的VEGFmRNA表达升高,提示其可能机制是通过VEGF途经来实现的。

AIM:To determine whether hydroxysafflor yellow A(HSYA) inhibits high glucose-induced cell proliferation and expression of vascular endothelial growth factor(VEGF) mRNA in retinal capillary endothelial cells(Macaca rhesus monkey vascular endothelial cells,RF/6A cells).METHODS:RF/6A cells cultured in vitro were divided into NG+H0 group in which contained with normal glucose of 5.5mol/L,and was added with HSYA of 0mg/L,and were divided into HG+H0,HG+H18,HG+H37,HG+H73 group,in which contained with high glucose of 22mmol/L and were added with HSYA of 0,18,37 and 73mg/L,respectively.At 24,48,72 hours cultured,the cells were observed by inverted microscope.The effects of HSYA on cell proliferation and expression of VEGF mRNA were tested by MTT assay and RT-PCR(reverse transcription-polymerase chain reaction),respectively.RESULTS:The results of MTT assay for the effects of HSYA on the proliferation of RF/6A cells were showed as follows:at 48hours of cells cultivation,compared with HG+H0,the effects in HG+H37 and HG+H73 group were statistically significant(P<0.01).With the concentration of HSYA increased in high glucose groups,the absorbance values of RF/6A cells showed a descending trend.At 72hours of cells cultivation,there were statistically significant effects in groups of HG+H18,HG+H37 and HG+H73 when compared with HG+H0 group(P<0.01).With the concentration of HSYA increased in high glucose groups,the absorbance values of RF/6A cells showed a descending trend,and there was a marked reduction in HG+H73 group compared with the HG+H18 and HG+H37 group(P<0.01,P<0.05,respectively).The RT-PCR results for the effects of HSYA on VEGF mRNA expression were showed as follows:at 48hours of cells cultivation,the expression of VEGF mRNA in HG+H37,HG+H73 group was statistically significant lower than that in HG+H0 group(P<0.01) and with the same results showed in HG+H18,HG+H37,HG+H73 group at 72hours of cells cultivation(P<0.01).CONCLUSION:These results suggested that HSYA can inhibit the expression of VEGF and the high glucose-induced proliferation of RF/6A cells.The inhibitory effect of cell proliferation was increased in both time-dependent and dose-dependent manner.These effects may be associated with down-regulating the expression of VEGF mRNA by HSYA.

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中国广西教育厅科研资助面上项目(No.200911MS25);中国广西大型仪器协作网资助项目(No.558-2008-001)~~