黄芪多糖对脂多糖诱导血管内皮细胞损伤的保护作用及相关机制的实验研究  

Experimental Study on Protective Effect and Mechanism of Astragalus Polysaccharide on Injury of Human Vascular Endothelial Cells Induced by LPS in Vitro

作　　者：秦劭晨 王爱梅 万晓波 QIN Shaochen;WANG Aimei;WAN Xiaobo(The Affiliated Hospital of Shanxi University of Chinese Medicine,Taiyuan,Shanxi 030024;Taiyuan Hospital of Chinese Medicine,Taiyuan,Shanxi 030009)

机构地区：[1]山西中医药大学附属医院,山西太原030024 [2]太原市中医医院,山西太原030009

出　　处：《中国中医药科技》2023年第1期37-41,共5页Chinese Journal of Traditional Medical Science and Technology

基　　金：国家区域中医医疗中心心血管专项基金项目(XGZX202117);山西省卫生健康委科研项目(2021038);山西中医药大学科技创新能力培育计划项目(2019PY-051)。

摘　　要：目的:研究黄芪多糖(Astragalus polysaccharide,APS)对脂多糖(LPS)诱导人血管内皮细胞(Human vascular endothelial cells,HUVECs)损伤的保护作用及机制。方法:以LPS(1 mg/L)诱导HUVECs损伤模型,取对数生长期的HUVECs,APS(0.5、1.0、2.0、4.0、8.0、16.0、32.0 mg/L)培养,采用CCK8检测各组细胞增殖活力。实验分正常组、LPS组、APS组(8.0 mg/L)和地塞米松(DEX)组(3.92 mg/L),LPS诱导细胞加入APS或DEX作用24 h,分光光度法检测一氧化氮(NO)、乳酸脱氢酶(LDH)和超氧化物歧化酶(SOD)表达;RT-PCR检测白细胞介素6(IL-6)、白细胞介素1β(IL-1β)、血管内皮生长因子(VEGF)、碱性成纤维生长因子(bFGF)mRNA表达;ELISA检测细胞上清IL-6、IL-1β、VEGF和bFGF水平;流式细胞术检测细胞凋亡情况。结果:与空白对照组相比,APS组可诱导HUVECs增殖;4~32 mg/L的APS对LPS诱导损伤模型细胞有保护作用,APS可以抑制HUVECs IL-6、IL-1β、VEGF、bFGF mRNA表达(P<0.01),下调NO和LDH水平,上调SOD水平(P<0.01),抑制HUVECs凋亡(P<0.01)。结论:APS可能通过下调IL-6、IL-1β、VEGF、bFGF mRNA表达、抑制氧化应激反应减轻LPS诱导的血管内皮细胞损伤。Objective:To investigate the protective effect of Astragalus(APS)polysaccharide on human vascular endothelial cells(HUVECs)injury induced by lipopolysaccharide(LPS)and explore its mechanism in vitro.Methods:HUVECs injury models were induced with LPS(1 mg/L),and HUVECs in logarithmic growth phase were cultured with APS(0.5,1.0,2.0,4.0,8.0,16.0,32.0 mg/L),CCK8 was used to detect the cells proliferation.Experiment was divided into normal group,LPS group,APS group(8 mg/L)and dexamethasone(DEX)group(3.92 mg/L),and LPS stimulated cells were added to APS or DEX effect for 24 h.Subsequently,CCK8 was used to detect the cells proliferation in each group;spectrophotometric assay was used to detect the levels of NO,LDH,SOD;the expressions of IL-6,IL-1β,VEGF and bFGF mRNA were determined by RT-PCR;the levels of IL-6,IL-1β,VEGF and bFGF in cell supernatant were detected by ELISA and cell apoptosis was detected by flow cytometry.Results:Compared with the control group,the APS groups of various concentrations could induced the proliferation of normal HUVECs,APS of 4~32 mg/L showed a protective effect on model of HUVECs injury induced by LPS in vitro.APS could inhibit the expressions of IL-6,IL-1β,VEGF,bFGF mRNA,down-regulate the levels of NO,LDH and up-regulate the level of SOD,inhibits apoptosis of HUVECs.Conclusion:APS may alleviate LPS-induced vascular endothelial injury by down-regulating IL-6,IL-1β,VEGF,bFGF levels and reducing oxidative stress.

关 键 词：黄芪多糖 血管内皮损伤 细胞增殖 一氧化氮 乳酸脱氢酶 超氧化物歧化酶 白细胞介素6 白细胞介素1Β 血管内皮生长因子 碱性成纤维生长因子 细胞凋亡 体外实验 

分 类 号：R285[医药卫生—中药学]