摘要目的 观察正常和缺氧条件下人参皂甙Rg3对人视网膜血管内皮细胞(HRCEC)增生、迁徙和管腔形成的影响.方法 在正常条件的培养基中,分别加入0.0、0.1、0.5 mmol/L的人参皂甙Rg3,依序分为A、B、C3个组；在150 μmol/L 氯化钴缺氧条件的培养基中,分别加入0.0、0.1、0.5 mmol/L的人参皂甙Rg3,依序分为D、E、F3个组.在培养后24、48、72 h用噻唑蓝比色法检测HRCEC的增生；在24 h,用Transwell小室检测细胞迁徙,用基质胶检测细胞管腔形成,实时荧光定量逆转录酶链聚合反应和蛋白质免疫印迹法检测细胞中血管内皮生长因子(VEGF)mRNA和蛋白质表达.结果 正常或缺氧条件培养基中,均可观察到人参皂甙Rg3对HRCEC增生的抑制作用,并且这种抑制作用呈浓度与时间依赖性(F正常组浓度=30.331,F缺氧组浓度=33.402,F正常组时间=85.462,F缺氧组时间=136.045,P均＜0.05).Transwell小室结果显示,A、B、C、D、E、F组HRCEC细胞的移行细胞数分别为(103.33±3.54)、(92.25±3.68)、(78.64±4.66)、(125.76±3.11)、(90.27±3.55)、(77.81±5.01)个.正常和缺氧条件下各组间HRCEC细胞的移行细胞数比较,差异有统计学意义(F正常组组间=28.801,F缺氧组组间=117.594,P均＜0.05).基质胶检测结果显示,A、B、C、D、E、F组HRCEC细胞的完整管腔形成数分别为(24.3±2.2)、(15.7±1.7)、(10.1±2.3)、(26.2±1.9)、(15.1±2.6)、(8.6±1.9)个.正常和缺氧条件下各组间HRCEC细胞的完整管腔形成数比较,差异有统计学意义(F正常组组间=35.364,F缺氧组组间=50.989,P均＜0.05).实时荧光定量逆转录-酶链聚合反应检测结果显示,A、B、C、D、E、F组HRCEC细胞的VEGF mRNA相对表达量分别为1.00±0.06、0.79±0.06、0.68±0.02、3.88±0.12、2.83±0.09、1.15±0.05.正常和缺氧条件下各组间HRCEC细胞的VEGF mRNA相对表达量比较,差异有统计学意义(F正常组组间=31.303,F缺氧组组间=682.668,P均＜0.05).蛋白质免疫印迹法检测结果显示,A、B、C、D、E、F组HRCEC细胞的VEGF蛋白质相对表达量分别为0.62±0.03、0.41±0.02、0.32±0.02、0.91±0.03、0.82±0.03、0.71±0.02.正常和缺氧条件下各组间HRCEC细胞的VEGF蛋白质相对表达量比较,差异有统计学意义(F正常组组间=125.471,F缺氧组组间=41.045,P均＜0.05).结论 人参皂甙Rg3可通过抑制HRCEC细胞增生、迁徙和管腔形成来抑制新生血管的形成,其机制可能通过抑制VEGF的表达来实现.

abstractsObjective To observe the effect of ginsenoside Rg3 on the proliferation,migration,and tube formation of human retinal capillary endothelial cell (HRCEC) cultured in normal and hypoxia condition.Methods HRCEC was cultured in normal condition and treated with 0.0 mmol/L (group A),0.1 mmol/L (group B) and 0.5 mmol/L (group C) ginsenoside Rg3.HRCEC was also cultured in hypoxia condition and treated with 0.0 mmol/L (group D),0.1 mmol/L (group E) and 0.5 mmol/L (group F)ginsenoside Rg3.The effects of ginsenoside Rg3 on HRCEC proliferation were measured by methylthiazoletrazolium assay in 24,48 and 72 hours after culture.In 24 hours after culture,the effect of cell migration was evaluated by transwell chamber; the effect of tube formation was evaluated by Matrigel; the expression of vascular endothelial growth factor (VEGF) protein and mRNA were detected by Western blot and real-time quantitative reverse transcription-polymerase chain reaction.Results Ginsenoside Rg3 could inhibit proliferation of HRCEC,depending on the concentration (F=30.331 and 33.402 in normal and hypoxia condition,respectively; P＜0.05) and time (F=85.462 and 136.045 in normal and hypoxia condition,respectively; P＜0.05).The number of cell migration was 103.33±3.54,92.25±3.68,78.64±4.66 in group A,B and C,the difference among three groups was statistically significant (F=28.801,P＜0.05).The number of cell migration was 125.76±3.11,90.27±3.55,77.81±5.01 in group D,E and F,the difference among three groups was statistically significant (F=117.594,P＜0.05).The number of tube formed in Matrigel was 24.3±2.2,15.7±1.7,10.1±2.3 in group A,B and C,the difference among three groups was statistically significant (F=35.364,P＜0.05).The number of tube formed in Matrigel was 26.2±1.9,15.1 ± 2.6,8.6 ± 1.9 in group D,E and F,the difference among three groups was statistically significant (F=50.989,P＜0.05).The expression of VEGF mRNA was 1.00±0.06,0.79±0.06,0.68±0.02 in group A,B and C,the difference among three groups was statistically significant (F=31.303,P＜0.05).The expression of VEGF mRNA was 3.88±0.12,2.83±0.09,1.15±0.05 in group D,E and F,the difference among three groups was statistically significant (F=682.668,P＜0.05).The expression of VEGF protein was 0.62±0.03,0.41±0.02,0.32±0.02 in group A,B and C,the difference among three groups was statistically significant (F=125.471,P＜0.05).The expression of VEGF protein was0.91±0.03,0.82±0.03,0.71±0.02 in group D,E and F,the difference among three groups was statistically significant (F=41.045,P＜0.05).Conclusion Ginsenoside Rg3 can inhibit the proliferation,migration,and tube formation of HRCEC through the inhibition of VEGF expression.