【摘要】 探究黄芪多糖（Astragalus polysaccharide, APS）是否通过调控VEGF/VEGFR影响大鼠肠黏膜微血管内皮细胞（rat intestinal mucosal microvascular endothelial cells, RIMMVECs）的增殖活性。从新生大鼠中分离RIMMVECs并进行纯化鉴定，随后稳定传代并以0.1、1.0、10.0、100.0、1 000.0和10 000.0 mg/L的APS处理RIMMVECs,应用MTT法研究APS对RIMMVECs增殖的影响并筛选APS最佳作用的质量浓度；利用流式细胞术检测细胞周期变化，评估APS对RIMMVECs细胞周期的作用阶段；用ELISA试剂盒检测细胞上清中VEGFA含量变化，评价细胞增殖诱导血管生成的潜力；用荧光显微镜观察Fluo-8AM负载的细胞内荧光强度变化，评估细胞内Ca²⁺水平；用蛋白免疫印迹（Western blot）法检测RIMMVECs中PERK蛋白表达水平，分析APS可能的作用机制。结果显示，100 mg/L APS显著增强RIMMVECs增殖活性，增加细胞上清中VEGFA含量，增加细胞Ca²⁺水平，升高细胞PERK蛋白表达量。表明APS具有促进RIMMVECs增殖的作用，该作用可能与促进细胞VEGFA表达以及激活ERK通路相关。更多还原

【Abstract】 This study explored whether Astragalus polysaccharide（APS） can regulate the VEGF/VEGFR signaling pathway to affect the proliferative activity of rat intestinal mucosal microvascular endothelial cells（RIMMVECs）.RIMMVECs were isolated from newborn rats,then purified and treated with APS at concentrations of 0.1,1.0,10.0,100.0,1 000.0,and 10 000.0 mg/L.MTT was used to determine the effect of APS on RIMMVECs proliferation and screen for the optimal concentration of APS.Subsequently,flow cytometry was used to detect the changes in cell cycle to evaluate the stage of action of APS on the cell cycle in RIMMVECs.Then,the ELISA was used to detect the changes of VEGFA in cell supernatant to evaluate the potential of cell proliferation and angiogenesis.The changes in fluorescence intensity of Fluo-8AM was observed using fluorescence microscopy to evaluate intracellular Ca²⁺ levels.Finally,Western blot was used to detect the expression of PERK in RIMMVECs to analyze the possible mechanism of APS.The results showed that 100 mg/L APS significantly enhanced the proliferative activity of RIMMVECs,increased the content of VEGFA in the cell supernatant,the intracellular Ca²⁺ levels,and the expression of PERK protein,indicating that APS promotes the proliferation of RIMMVECs,which may be achieved by promoting the expression of VEGFA and activating the ERK pathway.更多还原