摘要
目的探讨中药单体人参皂苷Rg3对血管内皮生长因子(VEGF)刺激下的猴脉络膜-视网膜内皮细胞(RF/6A)增殖与迁移能力的影响。方法利用不同浓度VEGF分别在不同时间干预刺激RF/6A细胞,采用CCK-8法检测细胞增殖情况,筛选VEGF最佳干预浓度及干预时间;应用不同浓度康柏西普干预VEGF刺激下的RF/6A细胞,观察筛选康柏西普最强作用浓度;应用低、中、高浓度人参皂苷Rg3和康柏西普干预VEGF刺激下的RF/6A细胞,采用CCK-8法检测各干预对细胞增殖的影响;采用细胞划痕实验检测人参皂苷Rg3和康柏西普对VEGF刺激下RF/6A细胞迁移能力的影响。结果CCK-8法检测结果显示,不同时间点,不同浓度VEGF刺激培养对细胞的增殖影响不同,其中,50 ng/mL VEGF刺激RF/6A细胞48 h后,细胞增殖率最大;不同浓度康柏西普干预结果显示,0.5μg/mL即能有效抑制VEGF刺激下RF/6A细胞的增殖(t=-3.147,P=0.035);低、中、高浓度人参皂苷Rg3及康柏西普组均能抑制VEGF诱导的RF/6A细胞增殖(F=26.546,P=0.000),其中,康柏西普组抑制作用强于人参皂苷Rg3各浓度组。细胞划痕实验各组的迁移率比较,干预培养24 h后,联合组对细胞移行的抑制作用强于人参皂苷Rg3组(t=4.639,P=0.010),康柏西普组的抑制作用亦强于人参皂苷Rg3组(t=4.350,P=0.012),而两组间比较无统计学意义(P>0.05);继续培养至48 h,康柏西普和人参皂苷Rg3组均能抑制细胞迁移,但差异无统计学意义(P>0.05),均低于联合组(t=8.543,P=0.001;t=3.067,P=0.037)。结论VEGF能够促进RF/6A细胞的增殖和迁移,中药单体人参皂苷Rg3和康柏西普均具有抑制这一增殖和迁移的作用,且联合应用抑制作用最强。
OBJECTIVE To investigate the effect of Chinese medicine Ginsenoside Rg3 on the proliferation and migration ability of monkey choroid-retinal endothelial cells(RF/6A)stimulated by vascular endothelial growth factor(VEGF).METHODS Different concentrations of VEGF were used to stimulate RF/6A cells at different times,and CCK-8 method was used to detect cell proliferation and optimal concentration and time of VEGF.Different concentrations of Conbercept were used to intervene in RF/6A cells stimulated by VEGF,and the best concentration of Conbercept was observed and screened.Low,medium and high concentrations of Ginsenoside Rg3 and Conbercept were used to intervene in RF/6A cells stimulated by VEGF,CCK-8 method was used to detect the effect of intervention on cell proliferation;Cell scratch test was used to detect the effect of ginsenoside Rg3 and Conbercept on migration of RF/6A cells stimulated by VEGF.RESULTS The results of CCK-8 method showed that at different time points and different concentrations of VEGF stimulated culture had different effects on cell proliferation.Among them,the cell proliferation rate was the largest,when 50 ng/mL VEGF stimulated RF/6A cells for 48 h;The results of different concentrations of Conbercept intervention showed that 0.5μg/mL could effectively inhibit the proliferation of RF/6A cells stimulated by VEGF(t=-3.147,P=0.035);Low,medium and high concentrations of Rg3 and Conbercept group could inhibit the proliferation of RF/6A cells induced by VEGF(F=26.546,P=0.000).And,the inhibitory effect of the Conbercept group was stronger than that of the Rg3 each concentration group.Comparison of the mobility of each group in the cell scratch test showed that after 24 h intervention culture,the combined group had a stronger inhibitory effect on cell migration than the Rg3 group(t=4.639,P=0.010;t=4.350,P=0.012),there was no statistically significant difference between the two groups(P>0.05);But continued to culture for 48 h,both the Conbercept and Rg3 groups could inhibit cell migration,there was no statistical significance(P>0.05),but all were lower than the combined group(t=8.543,P=0.001;t=3.067,P=0.037).CONCLUSIONS VEGF could promote the proliferation and migration of RF/6A cells.Both Ginsenoside Rg3 and Conbercept could inhibit the proliferation and migration of RF/6A cells,and the combined application had the strongest inhibition effect.
作者
褚文丽
陈水龄
亢泽峰
刘健
李维义
陶方方
CHU Wenli;CHEN Shuiling;KANG Zefeng(Eye Hospital,China Academy of Chinese Medical Sciences,Beijing 100040,China)
出处
《中国中医眼科杂志》
2020年第2期89-93,100,共6页
China Journal of Chinese Ophthalmology
基金
国家自然科学基金(81574032)
国家中医药管理局岐黄工程亢泽峰歧黄学者工作室项目
深圳市医疗卫生三名工程(SZSM201812090)
北京市石景山区亢泽峰名医传承工作室项目。
作者简介
通讯作者:亢泽峰,E-mail:Zefeng2531@163.com。
