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

Protective effects of total saponrins in Panax Notoginseng on high glucoseinduced injury in cultured rat retinal capillary endothelial cells( RRCECs)

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＆amp;nbsp;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.