摘要
目的:观察三七总皂苷( PNS)对高浓度葡萄糖致大鼠视网膜微血管内皮细胞( RRCECs)损伤的保护作用和对细胞黏附分子-1(ICAM-1)的影响。方法体外原代培养RRCECs,并通过免疫荧光化学方法鉴定;用PNS作用于4-6代的RRCECs,实验分为正常组、高糖组(30 mmol/L)和高糖不同浓度PNS(20、50、100 mg/L)组,分别使用MTT比色法和台盼蓝计数法观察PNS对细胞活力及数目的影响,使用Western blotting检测药物作用前后ICAM-1的表达变化。结果 MTT比色法结果显示,PNS 50、100 mg/L作用48 h和72 h可明显减轻高糖造成的RRCECs损伤,提高RRCECs活力(P<0.01);台盼蓝计数结果显示,在PNS作用48 h后PNS 100 mg/L组细胞数目明显增加(P<0.01),72 h后PNS 50、100 mg/L 组细胞数目均明显增加( P<0.05)。 Western blotting 检测显示,随着PNS药物浓度的增高,高糖环境下RRCECs中的ICAM-1表达量逐渐减少( P<0.05)。结论 PNS可以减轻高糖引起的RRCECs损伤,减少高糖环境下RRCECs中ICAM-1表达。 PNS对RRCECs的保护作用与ICAM-1表达调节之间的关系有待进一步研究。
Objective To observe the effects of total saponins of Panax Notoginseng ( PNS ) protection and expression of intercellular adhesion molecules-1 ( ICAM-1 ) in cultured rat retinal capillary endothelial cells ( RRCECs ) induced with high-concentration glucose.Methods Endothelial cells obtained from rat retinas were cultured in vitro, and the cells were identified using immunofluorescence. RRCECs were randomly divided into five groups: normal group, high glucose group, high glucose+20 mg/L PNS group, high glucose+50 mg/L PNS group, and high glucose+100 mg/L PNS group.Cell viability was measured with MTT method while cell number was detected with Trypan blue assay.The protein expression of ICAM-1 in RRCECs was measured by using Western blotting.Results According to MTT assay, the cell viability in high glucose+PNS (50 mg/L, 100 mg/L) groups were promoted after an action time of 48 hours and 72 hours (P〈0.01).According to Trypan blue assay, after 48 hours and 72 hours, the cell number in high glucose+PNS (100 mg/L) group was increased ( P〈0.01);after 48 hours, the cell number in high glucose+PNS (50 mg/L) group was increased (P〈0.05).After an action time of 72 hours, the expression of ICAM-1 in all high glucose+PNS groups were detected by using Western blotting.The result showed that the expression of ICAM-1 in RRCECs decreased with the rise of PNS concentration under high glucose condition ( P 〈0.01 ) .Conclusion PNS could protect RRCECs from damage caused by high glucose, and inhibit the expression of ICAM-1.Relation between protective effects of PNS and regulation of ICAM-1 expression in cultured RRCECs merits further study.
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2015年第5期318-322,I0001,共6页
Journal of Beijing University of Traditional Chinese Medicine
基金
"重大新药创制"国家科技重大专项(No.2011ZX09201-201-22)
北京中医药大学研究生自主课题(No.2014-JY13ZZ-XS-099)
关键词
三七总皂苷
视网膜微血管内皮细胞
细胞间黏附分子-1
大鼠
total Saponins of Panax Notoginseng
retinal capillary endothelial cells
intercellular adhesion molecules-1