血栓通胶囊对氧糖剥夺/复氧致HUVEC细胞紧密连接损伤的改善作用

目的采用氧糖剥夺(OGD)致HUVEC细胞损伤模型,考察三七总皂苷(PNS)及其单体Rg1、Rb1对氧糖剥夺致血管内皮紧密连接损伤的保护作用。方法 HUVEC细胞接种于Transwell细胞小室,待细胞融合后,设立实验组别:正常对照组、模型组、PNS 10、20、40 mg/L组以及Rb1 13.52 mg/L、Rg1 12.44 mg/L组,于缺氧同时进行药物干预,氧糖剥夺6 h复氧24 h后,检测HUVEC内皮细胞电阻值以及内皮渗透性的变化;采用激光共聚焦显微镜,免疫荧光法检测HUVEC细胞ZO-1、claudin-5蛋白表达的变化。结果与正常对照组相比,HUVEC细胞缺氧6 h复氧24 h后内皮细胞间紧密连接电阻显著降低,细胞渗透性显著升高(P0.05)。PNS 20、40 mg/L组能够显著抑制模型组紧密连接电阻的降低,抑制HUVEC细胞渗透性的升高(P0.05)。人参皂苷Rb1能够显著升高OGD 6 h复氧24 h后内皮紧密连接电阻,降低HUVEC细胞渗透性(P0.05)。免疫荧光检测结果显示,正常对照组HUVEC细胞的ZO-1及claudin-5蛋白主要分布于细胞膜周围,排列连续,显示出内皮细胞的典型铺路石排列。缺氧6 h复氧24 h损伤后,模型组细胞边缘的紧密连接蛋白显著减少,环形结构断裂,甚至消失,细胞核内紧密连接蛋白呈增加趋势。PNS 20、40 mg/L及Rb1 13.52 mg/L可观察到细胞间紧密连接局部恢复,呈铺路石样结构。结论 PNS对缺血致血管内皮细胞间紧密连接损伤有保护作用。

Xueshuantong capsule alleviates oxygen-glucose deprivation / reoxygenation-induced tight junction damage of HUVEC cells

Objective To study the protective effect of Panax notoginseng saponins (PNS) and its components Rg1 and Rb1 on oxygen-glucose deprivation/ reoxygenation ( OGD/ Reox)-induced tight junction damage. Methods Anaerobic box were used to induce OGD in HUVEC cells for 6 h followed by reoxygenation for 24 h. Transepithelial/endothelial electrical resistance (TEER) and cell permeability were detected, immunefluorescence was used to observe the ZO-1 and claudin-5 protein expression. Results PNS 20, 40 mg/ L and ginsenoside Rb1 significantly inhibited the OGD/Reox-induced decreased tight junction resistance, and the increased cell permeability (P < 0.05). PNS 20, 40 mg/ L and ginsenoside Rb1 partly restored the inter-cellular tight junctions which were regularly arranged on the cell membrane, and the cells displayed cobble stone-like arrangement. Conclusions PNS ameliorates ischemia-induced vascular endothelial cell tight junction damage via MMP-9 and VEGF/ VEGFR2 signaling pathway. Rb1 is one of the effective monomer components.