摘要
目的分别从体外实验和体内实验两个方面探讨人参皂苷RG3对脉络膜新生血管的抑制作用。方法体外实验:MTT检测人参皂苷RG3对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)增殖的抑制作用,HUVEC分为正常组(含体积分数10%胎牛血清的DMEM/F^(-1)2培养基)、对照组(含2 g·L^(-1)DMSO)和12.5μmol·L^(-1)、25.0μmol·L^(-1)、50.0μmol·L^(-1)、100.0μmol·L^(-1)人参皂苷RG3给药组,给药处理6 h后测定其吸光度值。HUVEC分为对照组(含2 g·L^(-1)DMSO)和100.0μmol·L^(-1)人参皂苷RG3组,小管形成实验检测人参皂苷RG3对小管形成的抑制作用,Western blotting检测人参皂苷RG3对VEGF蛋白表达的影响。体内实验:雄性C57BL/6J小鼠20只,随机分为对照小鼠组和人参皂苷RG3小鼠组,皮下注射预处理给药2周,半导体激光造模,于光凝后第1天和第21天行荧光素眼底血管造影检查。结果 MTT结果显示,正常组、对照组、12.5μmol·L^(-1)、25.0μmol·L^(-1)、50.0μmol·L^(-1)、100.0μmol·L^(-1)人参皂苷RG3给药组的吸光度值分别为0.43±0.17、0.43±0.05、0.33±0.02、0.24±0.02、0.18±0.01、0.15±0.01,对照组与正常组相比差异无统计学意义(P>0.05),其余各组与对照组相比差异均有统计学意义(均为P<0.05)。小管形成实验结果显示,对照组与100.0μmol·L^(-1)人参皂苷RG3组小管形成数目分别为(72.5±5.56)个和(11.33±3.71)个,差异有统计学意义(P<0.01)。Western blotting结果显示,100.0μmol·L^(-1)人参皂苷RG3组的VEGF蛋白相对表达量(0.14±0.01)明显低于对照组(0.46±0.01),差异有统计学意义(P<0.01)。体内实验荧光素眼底血管造影结果显示,人参皂苷RG3小鼠组的荧光渗漏面积明显低于对照小鼠组。结论人参皂苷RG3可以抑制脉络膜新生血管的形成。
Objective To observe the inhibitory effect of ginsenoside RG3 on choroidal neovascularization in vitro and in vivo. Methods MTT assay was used to evaluate the inhibitory effect of ginsenoside RG3 on human umbilical vein endothelial cells （HUVEC） proliferation in vitro, and HUVEC were divided into normal group, in which the cells were cultured in DMEM/F-12 medium containing 10% fetal bovine ser- um, control group with its medium containing 2 g ·L-1 dimethyl sulfoxide （DMSO） and different concentrations of ginsenoside RG3 administration group （12. 5 pμmol ·L-1, 25.0 μmol·L-1 ,50.0 μmol ·L-1 ,100.0 μmol·L-1 ）. Then the absorbance value was measured after 6 h. Then, a small amount of HUVEC was collected again and divided in- to control group with its medium containing 2 g ·L-1 DMSO and 100.0 ixmol ·L-1 gin- senoside RG3 group for detecting the inhibitory effect of ginsenoside RG3 on tubular formation and vascular endothelial growth factor （VEGF） protein expression by West- ern blots. In vivo, 20 male C57BL/6J mice were collected and randomly divided into control group and glnsenoside RG3 group. After 2 weeks, followed by establishment of model with a semiconductor laser, fundus fluorescein anglography was performed on the 1st day and 21 st days after treatment. Results MTT results showed that absor- bance value of the normal group,control group,12.5 μmol ·L-1 ,25.0 μmol ·L-1 ,50.0 txmol·L-1 ,100.0 iμmol ·L-1 ginsenoside RG3 group was 0.43 ±0. 17,0.43 ＋0.05, 0.33 ± 0.02,0.24 ± 0. 02,0.18 ± 0.01,0. 15 ± 0.01 accordingly, and there was no signifi- cant difference between the control group and the normal group （ all P 〉 0.05 ）, but the difference between the other group and control group was statistically significant （all P 〈 0.05 ）. Tubular formation assay showed that the number of tubular formation in the control group and 100. 0 μmol ·L-1 ginsenoside RG3 group was 72. 5 ＋ 5. 56 and l l. 33 3.71, respectively, and the difference was statistically significant （P 〈 0.01 ）. Western blots showed that the relative expression of VEGF in 100.0 μmol·L-1 ginsen- oside RG3 group （0.14 ± 0.01 ） was significantly lower than that in the control group （0.46 ±0. 01 ） ,and the difference was statistically significant （P 〈0.01 ）. In vivo fundus fluorescein angiography showed that the fluorescein leakage area of ginsenoside RG3 group was lower than that of the control group. Coneluslon Ginsenosid RG5 can in-hibit the tormation of choroidal neovascularization by inhibiting the expression of VEGF in vitro and in vivo.
出处
《眼科新进展》
CAS
北大核心
2017年第10期922-925,共4页
Recent Advances in Ophthalmology
基金
辽宁省科学技术基金[面上项目(原优秀人才培育)](编号:201602298)
辽宁省科技厅-锦州医科大学联合基金项目(编号:20170540382)~~
