洋参二醇皂苷对波动高糖诱导人脐静脉内皮细胞损伤的保护效应研究

摘要

目的：探讨洋参二醇皂苷对波动高糖诱导人脐静脉内皮细胞损伤的保护效应及其机制。方法以体外培养人脐静脉内皮细胞为研究对象，通过每24 h间替更换高糖型DMEM全培养基（葡萄糖浓度25 mmol／L）与低糖型DMEM全培养基（葡萄糖浓度5．56 mmol／L）构建人脐静脉内皮细胞血糖波动模型。实验分为5组，包括正常低糖组、稳定高糖组、波动高糖组（间替高糖／低糖）、洋参二醇皂苷低剂量组（0．04 mg／mL）、洋参二醇皂苷高剂量组（0.08 mg／mL）。细胞培养8d ，观察细胞形态学表现，MTT测定细胞成活率，取细胞培养上清液检测乳酸脱氢酶（ LDH）、一氧化氮（NO）、内皮素－1（ET－1）、可溶性细胞间黏附分子－1（sICAM－1）、肿瘤坏死因子－α（TNF －α）含量，细胞裂解液检测胞内超氧化物歧化酶（ SOD）活性和丙二醛（ MDA）含量。结果洋参二醇皂苷低、高剂量组细胞形态均较波动高糖组明显好转，细胞活性和细胞中SOD活性均明显高于波动高糖组（P均＜0．05），LDH漏出率和细胞中TNF－α、sICAM－1、ET－1、NO、MDA 含量均明显低于波动高糖组（P均＜0．05）。结论洋参二醇皂苷对波动高糖诱导的人脐静脉内皮细胞具有显著保护效应，其保护效应与其能够显著减轻葡萄糖波动诱导的氧化应激、炎症反应及细胞黏附分子的过度表达密切相关。%Objective It is to observe the protection of Panax quinquefolium diol saponi ( DS) on human umbilical vein en-dothelial cells(HUVECs) suffering fluctuant high glucose and investigate the relative mechanism .Methods HUVECs were used as the research objects .Fluctuant high glucose was induced by a daily alternating 5.56 or 25 mmol/L glucose.There were five groups:normal glucose(NOR), steady high glucose(SHG), fluctuant high glucose(FHG ), low DS group(0.04 mg/mL,DSL),high DS group(0.08 mg/mL,DSH).HUVECs were incubated for 8 days.On the 9th day, the morphological changes were observed by light and electron microscopy , cell viability was measured by MTT method , cell membrane damage was determined by lactate dehydrogenase (LDH) leakage, the NO, ET-1, TNF-α, sICAM-1 concentrations in the cell culture supernatant were measured by ELISA method .The activities of SOD and the contents of MDA in the cell lysate were examined by enzyme method or spectrophotometry .Results Morphological results showed that both DSL and DSH cloud alleviate HU-VECs injury induced by high blood glucose fluctuation obviously .The cell survival rates and cell viability were all significantly increased compared with FHG group (P<0.05).In comparison with FHG group, the levels of NO, MDA, ET-1, TNF-α, sICAM-1 in DSL and DSH groups were all significantly decreased (P<0.05), meanwhile the level of SOD was increased sig-nificantly(P<0.05).Conclusion Panax quinquefolium diol saponin (DS) can protect HUVECs suffering fluctuant high glu-cose obviously .The protective mechanism is closely related to its effects of relieving vessel oxidative stress , alleviating inflam-matory reaction and decreasing the expressions of cellular adhesion molecule .