三七总皂苷对高糖诱导大鼠视网膜微血管内皮细胞损伤的保护作用

摘要

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.%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。