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目的考察不同相对分子质量岩藻聚糖硫酸酯对内皮细胞的保护作用并探讨其机制。方法采用体内研究和体外研究的方法,探讨不同相对分子质量海带岩藻聚糖硫酸酯对内皮细胞的保护作用。在体内研究中,通过制作大鼠内皮损伤模型,显微镜下观察大鼠胸主动脉内皮损伤情况,并测定内皮损伤指标血管性血友病因子(vWF)。在体外研究中,通过细胞培养,以血管性血友病因子和内皮微颗粒(EMPs)水平作为内皮损伤指标,考察药物对内皮损伤的保护作用。结果体内研究:光学显微镜下HE切片观察显示,相对低相对分子质量组(LF1组和LF2组)对内皮细胞的保护作用强于相对中相对分子质量组(MFa组和MFb组)。与模型组比较,LF1和LF2可显著降低血管性血友病因子水平(P<0.05),而MFa和MFb与模型组无显著性差异(P>0.05)。体外研究:与模型组比较,LF1和LF2可显著降低血管性血友病因子水平(P<0.05),而MFa和MFb与模型组无显著性差异(P>0.05)。LF1和LF2组的内皮微颗粒数目显著低于模型组(P<0.01),而与正常对照组无统计学差异;MFa和MFb组的内皮微颗粒数目比正常对照组明显增多(P<0.01),数目虽比模型组有所减少,但无统计学差异(P>0.05)。结论低相对分子质量的岩藻聚糖硫酸酯对内皮细胞均具有良好的保护作用,其通过降低血管性血友病因子和内皮微颗粒浓度,阻断血小板与受损血管内皮中的胶原结合,从而抑制了血小板活化,发挥内皮保护作用;而相对中相对分子质量的岩藻聚糖硫酸酯对内皮细胞的损伤改善不明显。
Objective To investigate the protective effect of fucoidan on the endothelial cells with different molecular weight and explore its mechanism. Methods In vivo and in vitro studies were conducted to investigate the protective effects of fucoidan on the endothelial cells with different relative molecular weights. In the in vivo study, the rat model of endothelial injury was made and the endothelium injury of the thoracic aorta was observed under a microscope. The vWF was measured. In vitro studies, through the cell culture, von Willebrand factor and endothelial micro-particles (EMPs) level as an indicator of endothelial damage, study the drug’s protective effect on endothelial injury. Results In vivo studies: HE sections under light microscopy showed that the protective effect of the relative low molecular weight group (LF1 and LF2) on endothelial cells was stronger than that of the relative mid-molecular weight group (MFa and MFb). Compared with the model group, LF1 and LF2 could significantly reduce the level of vWF (P <0.05), while MFa and MFb had no significant difference with the model group (P> 0.05). In vitro studies: Compared with the model group, LF1 and LF2 significantly decreased the levels of vWF (P <0.05), whereas MFa and MFb did not differ significantly from the model group (P> 0.05). The number of endothelial micro-particles in LF1 and LF2 group was significantly lower than that in model group (P <0.01), but not in normal control group. The number of endothelial micro-particles in MFa and MFb group was significantly higher than that in normal control group (P <0.01) , Although the number was reduced than the model group, but no significant difference (P> 0.05). Conclusions Fucoidan sulfate with low relative molecular mass has a good protective effect on endothelial cells by blocking the concentration of von Willebrand factor and endothelial micro-particles, blocking the platelets from collapsing in the damaged vascular endothelium, Thereby inhibiting platelet activation, exerting endothelial protection; while the relative molecular weight of fucoidan sulfate damage to endothelial cells did not improve significantly.