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[摘要]
目的:研究青蒿琥酯(artesunate)对体外高糖环境下培养的人视网膜微血管内皮细胞(human retinal capillary endothelial cells, HRCECs)的增殖及血管内皮细胞生长因子(vascular endothelial growth factor,VEGF)表达的影响。
方法:从新鲜人眼球提取HRCECs,进行体外培养。取第3~4代生长良好的细胞用于实验,实验分为低糖对照组、高糖对照组、高糖+不同浓度青蒿琥酯组(15,30,60,120μg/mL),用噻唑蓝(MTT)比色法检测HRCECs的增殖,通过免疫细胞化学法观察各分组HRCECs中VEGF的表达。
结果:MTT比色法检测细胞增殖结果显示:与低糖组比较,高糖显著促进HRCECs增殖(P>0.05); 15,30,60,120μg/mL青蒿琥酯处理24h或48h,可时间和浓度依赖性地抑制高糖环境下HRCECs的增殖(P<0.05)。免疫细胞化学法检测VEGF表达结果显示:与低糖对照组相比,高糖对照组VEGF表达明显增加(P<0.05); 高糖+不同浓度青蒿琥酯(15,30,60μg/mL)组与高糖对照组相比VEGF表达明显下降(P<0.05),高糖+不同浓度青蒿琥酯组之间两两比较均具有统计学差异(P<0.05),且随着药物浓度的提高,VEGF的表达量逐渐减少。
结论:青蒿琥酯呈浓度依赖性地抑制高糖环境下HRCECs的增殖以及VEGF的表达,表明青蒿琥酯抑制高糖环境诱导HRCECs增殖可能与其抑制VEGF表达有关。
[Key word]
[Abstract]
AIM: To study the effect of artesunate(ART)on proliferation and expression of vascular endothelial growth factor(VEGF)in cultured human retinal capillary endothelial cells
in vitro under the condition of high glucose.
METHODS: Human retinal capillary endothelial cells(HRCECs)extraction was taken from fresh human eyeand cultured in vitro. The cells of good growth in 3rd-4th generation were used into the experiment. The experimental subjects were divided into low glucose control group, high glucose control group and high glucose+different concentrations ART(15μg/mL, 30μg/mL, 60μg/mL, and 120μg/mL)groups. The effect of ART on the proliferation of HRCECs for 24 and 48 hours was measured by MTT assay. The effect of ART on the expression of VEGF in HRCECs was detected by immunocytochemistry.
RESULTS: The results from MTT assay showed that high glucose significantly increased the proliferation of HRCECs(P>0.05)and treatment of HRCECs with ART at the concentrations 15μg/mL,30μg/mL,60μg/mL,120μg/mL for 24 and 48 hours caused a time- and dose-dependent inhibition in the proliferation of HRCECs(P<0.05). The expression of VEGF in high glucose group was significantly higher than that in the low glucose control group(P<0.05). Compared with the high glucose control group, the expressions of VEGF in high glucose + different concentrations ART(15μg/mL,30μg/mL,60μg/mL)groups were decreased in a dose-dependent manner(P<0.05).
CONCLUSION: ART concentration-dependently inhibits high glucose-induced proliferation and VEGF overexpression in HRCECs, indicating that ART-inhibited the proliferation of HRCECs exerted by high glucose environment might be related to the inhibition of VEGF expression.
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