阿司匹林对丙酮醛引起的人血脑屏障内皮细胞损伤的影响  

Effect of aspirin on the injury of hCMEC/D3 cells caused by methylglyoxal

作　　者：庄世芳 王艳华 ZHUANG Shi-fang;WANG Yan-hua(Department of Clinical Laboratory,Xi'an Ninth Hospital Affiliated to Medical College of Xi'an Jiaotong University,Xi'an 710054,Shaanxi,CHINA;Department of Pathology,Xi'an Ninth Hospital Affiliated to Medical College of Xi'an Jiaotong University,Xi'an 710054,Shaanxi,CHINA)

机构地区：[1]西安交通大学附属西安市第九医院检验科,陕西西安710054 [2]西安交通大学附属西安市第九医院病理科,陕西西安710054

出　　处：《海南医学》2020年第2期141-145,共5页Hainan Medical Journal

基　　金：陕西省西安市卫生局科研项目(编号:J201701001)

摘　　要：目的探讨阿司匹林对丙酮醛(MGO)引起的人血脑屏障内皮细胞(hCMEC/D3)损伤的影响及调控机制。方法首先,观察WST-1试剂盒检测含不同浓度MGO(0.1~10 mmol/L)或阿司匹林(0.1~1 mmol/L)的培养液处理24 h对细胞活力的影响。然后,将实验细胞分组处理:对照组(培养液加入对应体积的DMSO或去离子水)、MGO处理组(培养液加入1 mmol/L的MGO)、MGO联合阿司匹林处理组(培养液加入1 mmol/L的MGO和不同浓度的阿司匹林),培养24 h后,试剂盒检测细胞活力、Caspase-3活性、活性氧(ROS)水平,Western blot检测LC3Ⅰ、LC3Ⅱ表达情况。结果不同浓度的MGO处理细胞24 h后,细胞活力随MGO浓度提高而逐步降低,当MGO浓度达到1 mmol/L时,细胞活力为(79.72±5.01)%,与0 mmol/L MGO组的(100.00±2.45)%比较差异有统计学意义(P<0.05);不同浓度的阿司匹林处理细胞24 h后,阿司匹林浓度依赖性降低细胞活力,当阿司匹林浓度为0.5 mmol/L或1 mmol/L时,细胞活力为分别为(87.72±3.36)%和(81.55±2.85)%,与0 mmol/L阿司匹林组相比,差异有统计学意义(P<0.05);MGO联合阿司匹林处理进一步降低细胞活力,MGO联合0.5 mmol/L或1 mmol/L阿司匹林处理组,细胞活力分别为(67.33±6.47)%和(59.40±5.12)%,与MGO处理组的(80.16±3.50)%相比,差异均有统计学意义(P<0.05);MGO处理组的Caspase-3活性为(112.38±4.62)%,较对照组的(100.00±2.32)%有所升高,但差异无统计学意义(P>0.05);MGO联合1 mmol/L阿司匹林处理组,Caspase-3活性为(133.13±7.23)%,与对照组或MGO处理组相比,差异均具有统计学意义(P<0.05);与对照组相比,MGO处理组ROS阳性的细胞数明显增加,差异具有统计学意义(P<0.05);MGO联合不同浓度阿司匹林处理组ROS阳性的细胞数较MGO处理组均进一步增加,差异有统计学意义(P<0.05)。与对照组相比,MGO处理组LC3Ⅱ/LC3Ⅰ表达比值上升至4.7倍,然而阿司匹林能够完全抑制MGO引起的LC3Ⅱ表达增加。结论阿司匹林加重丙酮醛引起Objective To study the effects of aspirin on methylglyoxal(MGO) induced injury in human blood-brain barrier model cell lines(hCMEC/D3) and its regulatory mechanism. Methods First, WST-1 kit was used to detect the effect of different concentrations of MGO(0.1 mmol/L to 10 mmol/L) or aspirin(0.1 mmol/L to 1 mmol/L)on cell viability after 24 h treatment. Then, the cells were divided into three groups: control group(culture solution with DMSO or deionized water of corresponding volume added), MGO treatment group(culture solution with 1 mmol/L MGO added), MGO combined with aspirin treatment group(culture solution with 1 mmol/L MGO and aspirin of different concentrations added). After cultured for 24 h, the cell activity, Caspase-3 activity, and ROS level were detected by kits and the expression of LC3Ⅰ and LC3Ⅱ were detected by Western blot. Results After treatment with MGO(0.1 mmol/L to 10 mmol/L) for 24 h, cell viability decreased gradually with the increase of MGO concentration. When MGO concentration reached 1 mmol/L, cell viability was(79.72±5.01)%, which was significantly lower than(100.00±2.45)% of control group(P<0.05). After treatment with aspirin of different concentrations for 24 h, aspirin concentration-dependent decreased cell viability. When aspirin concentration was 0.5 mmol/L or 1 mmol/L, the cell viability was(87.72±3.36)% and(81.55±2.85)% respectively, which was significantly lower than that of 0 mmol/L aspirin group(P<0.05). MGO combined with aspirin treatment further reduced cell viability. The cell viability of MGO combined with 0.5 mmol/L or 1 mmol/L aspirin treatment group was(67.33±6.47)% and(59.40±5.12)% respectively, and the difference was statistically significant compared with(80.16±3.50)% of MGO treatment group(P<0.05). The Caspase-3 activity of MGO treatment group was(112.38±4.62)%, which was not significantly different from(100.00±2.32)% of control group(P<0.05). The Caspase-3 activity of MGO combined with 1 mmol/L aspirin treatment group was(133.13 ±7.23)%, which was signi

关 键 词：阿司匹林 人血脑屏障内皮细胞 丙酮醛 活性氧 自噬 凋亡 

分 类 号：R972[医药卫生—药品]