七叶皂苷钠可通过上调水通道蛋白-4的表达抑制脂多糖诱导的血管内皮炎症损伤  

Aescinate alleviates lipopolysaccharide-induced inflammatory injury in endothelium by up-regulation of aquaporin-4

作　　者：李孟垚 费瑞[2] 韩冬梅[1] 戚少龙 王欣宇 李伟业 杜建时[1] Li Mengyao;Fei Rui;Han Dongmei;Qi Shaolong;Wang Xinyu;Li Weiye;Du Jianshi(Department of Lymphatic Vascular Surgery,China-Japan Union Hospital of Jilin University,Key Laboratory of Lymphatic Surgery Jilin Province,Engineering Laboratory of Lymphatic Surgery Jilin Province,Changchun 130031,China;Department of Cell Biology,School of Basic Medical Sciences,Jilin University,Changchun 130031,China;Department of Chemistry,Tsinghua University,Beijing 100084,China)

机构地区：[1]吉林大学中日联谊医院淋巴血管外科,吉林省淋巴外科重点实验室,吉林省淋巴外科工程实验室,长春130031 [2]吉林大学基础医学院细胞生物学系,长春130021 [3]清华大学化学系,北京100084

出　　处：《中华实验外科杂志》2021年第12期2393-2396,共4页Chinese Journal of Experimental Surgery

摘　　要：目的探讨七叶皂苷钠(SA)对脂多糖(LPS)诱导的血管内皮损伤致炎症作用及机制。方法IMDM培养基(含10%胎牛血清)体外培养人脐静脉内皮细胞(HUVEC),用细胞计数试剂盒(CCK-8)检测0.4、1.6和6.4μg/ml的SA对HUVEC的毒性,采用2.5μg/ml和作用时间为6 h的LPS构建炎症损伤细胞模型。实验分组为空白组、模型组(2.5μg/ml LPS)、低剂量组(2.5μg/ml LPS+0.4μg/ml SA)、中剂量组(2.5μg/ml LPS+1.6μg/ml SA)和高剂量组(2.5μg/ml LPS+6.4μg/ml SA)。酶联免疫吸附试验(ELISA)检测各组细胞白细胞介素(IL)-6的表达水平;实时定量反转录聚合酶链反应(RT-qPCR)检测各组细胞中水通道蛋白-4(AQP4)的mRNA表达量;蛋白质印迹法(Western blot)和免疫荧光检测各组细胞中AQP4蛋白表达量。两组间比较采用非成对t检验,多组间比较采用单因素方差分析。结果CCK-8检测结果表明,0.4、1.6和6.4μg/ml的SA作用6 h后的细胞活性百分数与空白组比较,差异无统计学意义(91.867±3.758、93.695±4.638、95.838±5.558比100.000±0.000,F=2.946,P>0.05)。IL-6的ELISA检测结果显示,低、中、高剂量组中IL-6的相对表达量均低于模型组(1.224±0.017、1.119±0.155和1.009±0.012比1.281±0.075,F=5.673,P<0.05);RT-qPCR结果显示,低、中、高剂量组中AQP4 mRNA的相对表达量均高于模型组(0.770±0.013、0.854±0.014和0.895±0.014比0.713±0.015,F=102.237,P<0.01);Western blot结果显示,低、中、高剂量组中AQP4蛋白相对表达量均高于模型组(0.758±0.084、0.774±0.079和0.925±0.033比0.668±0.132,F=4.283,P<0.05)。结论SA能抑制LPS诱导的HUVEC损伤及炎性因子表达,其机制很可能与上调细胞内AQP4表达有关。Objective To investigate the anti-inflammatory effects and underlying mechanisms of sodium aescinate(SA)against lipopolysaccharide(LPS)induced inflammation injury in endothelium.Methods Human umbilical vein endothelial cell(HUVEC)cells cells were cultured by IMDM medium containing 10%fetal bovine serum and stimulated with various concentrations(0.4,1.6 and 6.4μg/ml)of SA.The toxicity of SA on HUVECs was determined by cell counting kit-8(CCK-8).The cells were treated with LPS(2.5μg/ml)for 6 h to establish an inflammatory cell model.According to different treatments,HUVEC cells were divided into the control group,the model group(2.5μg/ml LPS),and low(2.5μg/ml LPS+0.4μg/ml SA),medium(2.5μg/ml LPS+1.6μg/ml SA)and high(2.5μg/ml LPS+6.4μg/ml SA)dose of SA groups.Enzyme linked immunosorbent assay(ELISA)was used to detect the expression of interleukin(IL)-6.The mRNA expression of aquaporin-4(AQP4)in each group was detected by real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR).Immunofluorescent staining and Western blottin analysis were applied to investigate the protein expression of AQP4.The comparison between the two groups was done by the non-paired t test,and the comparison among multiple groups was carried out by the multi-factor variance analysis.Results The CCK-8 assay results showed that there was no statistically significant difference in SA groups(0.4,1.6 and 6.4μg/ml)compared with the control group(91.867±3.758,93.695±4.638,95.838±5.558 vs.100.000±0.000,F=2.946,P>0.05).ELISA results demonstrated that compared with the model group,the IL-6 expression of low-,medium-and high-dose groups were lower(1.224±0.017,1.119±0.155,1.009±0.012 vs.1.281±0.075,F=5.673,P<0.05).RT-qPCR analysis showed that compared with model group,the mRNA expression of AQP4 in low-,medium-and high-dose groups increased(0.770±0.013,0.854±0.014,0.895±0.014 vs.0.713±0.015,F=102.237,P<0.01).Western blotting indicated that the protein expression of AQP4 was higher in low-,medium-and high-dose gro

关 键 词：七叶皂苷钠 水通道蛋白4 血管内皮细胞 炎症 

分 类 号：R285[医药卫生—中药学]