丁苯酞对Aβ1-42的诱导HBMEC细胞凋亡及TLR4/COX-2表达的影响
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摘要
目的:研究丁苯酞对Aβ1-42诱导的HBMEC细胞凋亡的影响以及TLR4/COX-2的表达情况。方法:将HBMEC分为阴性对照组、Aβ1-42组、TAK242组(10 nmol/L)、DMSO组(1‰)和丁苯酞组(160、80、40μg/L)。采用CCK-8法检测细胞活力,流式细胞仪检测细胞凋亡率,免疫印迹法检测细胞中TLR4、COX-2蛋白的表达情况。结果:同阴性对照组比较,Aβ1-42组细胞早期凋亡率上升,TLR4、COX-2蛋白表达增加;与Aβ1-42组比较,TAK242组和丁苯酞组细胞凋亡率降低,TLR4、COX-2蛋白表达减少,比较结果有统计学意义(P<0.05)。结论 :丁苯酞对Aβ1-42所致的HBMEC细胞凋亡有保护效应,能降低细胞TLR4、COX-2蛋白的表达,其保护效应可能与抑制细胞TLR4、COX-2蛋白的表达有关。
Objective: To investigate the effects of butylphthalide on Aβ1-42-induced apoptosis and the expression of Toll-like receptor 4/cyclooxygenase-2(TLR4/COX-2) in human brain microvascular endothelial cells(HBMECs). Methods: HBMECs were divided into negative control group, Aβ1-42 group, TAK242(TLR4 inhibitor) group(10 nmol/L), dimethylsulfoxide(DMSO) group(1‰), and butylphthalide group(160 μg/L, 80μg/L, and 40 μg/L). Cell viability was determined by Cell Counting Kit-8, the apoptosis rate was determined by flow cytometry, and the expression of TLR4 and COX-2 in the cells was determined by Western blot. Results:Compared with the negative control group, the Aβ1-42 group had higher early apoptosis rate and expression of TLR4 and COX-2. Compared with the Aβ1-42 group, the TAK242 group and the butylphthalide group had significantly lower apoptosis rate and expression of TLR4 and COX-2(P < 0.05). Conclusions: Butylphthalide has a protective effect on HBMECs apoptosis induced by Aβ1-42 and can reduce the expression of TLR4 and COX-2 in HBMECs. Its protective effect may be related to the inhibition of TLR4 and COX-2 expression in HBMECs.
Objective: To investigate the effects of butylphthalide on Aβ1-42-induced apoptosis and the expression of Toll-like receptor 4/cyclooxygenase-2(TLR4/COX-2) in human brain microvascular endothelial cells(HBMECs). Methods: HBMECs were divided into negative control group, Aβ1-42 group, TAK242(TLR4 inhibitor) group(10 nmol/L), dimethylsulfoxide(DMSO) group(1‰), and butylphthalide group(160 μg/L, 80μg/L, and 40 μg/L). Cell viability was determined by Cell Counting Kit-8, the apoptosis rate was determined by flow cytometry, and the expression of TLR4 and COX-2 in the cells was determined by Western blot. Results:Compared with the negative control group, the Aβ1-42 group had higher early apoptosis rate and expression of TLR4 and COX-2. Compared with the Aβ1-42 group, the TAK242 group and the butylphthalide group had significantly lower apoptosis rate and expression of TLR4 and COX-2(P < 0.05). Conclusions: Butylphthalide has a protective effect on HBMECs apoptosis induced by Aβ1-42 and can reduce the expression of TLR4 and COX-2 in HBMECs. Its protective effect may be related to the inhibition of TLR4 and COX-2 expression in HBMECs.
引文
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11 .Gambuzza ME,Sofo V,Salmeri FM,Soraci L,et al.Tolllike receptors in Alzheimer’s disease:a therapeutic perspective[J].CNS Neurol Disord Drug Targets,2014,13(9):1 542-1 558.
12 .齐凡星,胡莹,卢军栋,等.丁苯酞治疗阿尔茨海默病的临床观察[J].中国药房,2016,27(17):2 412-2 414.
2 .Sun X,Chen WD,Wang YD.β-Amyloid:the key peptide in the pathogenesis of Alzheimer’s disease[J].Front Pharmacol,2015,6:221.
3 .Gold M,El Khoury J.β-amyloid,microglia,and the inflammasome in Alzheimer’s disease[J].Semin Immunopathol,2015,37(6):607-611.
4 .Huang NQ,Jin H,Zhou SY,et al.TLR4 is a link between diabetes and Alzheimer’s disease[J].Behav Brain Res,2017,316:234-244.
5 .Ma JF,Wang HM,Li QY,et al.Starvation triggers Abeta42 generation from human umbilical vascular endothelial cells[J].FEBS Lett,2010,584(14):3 101-3 106.
6 .Zhang W,Wang LZ,Yu JT,et al.Increased expressions of TLR2 and TLR4 on peripheral blood mononuclear cells from patients with Alzheimer’s disease[J].J Neurol Sci,2012,315(1-2):67-71.
7 .Gambuzza ME,Sofo V,Salmeri FM,Soraci L,et al.Tolllike receptors in Alzheimer’s disease:a therapeutic perspective[J].CNS Neurol Disord Drug Targets,2014,13(9):1 542-1 558.
8 .Shi S,Liang D,Chen Y,et al.Gx-50 reducesβ-amyloid-induced TNF-α,IL-1β,NO,and PGE2 expression and inhibits NF-κB signaling in a mouse model of Alzheimer's disease[J].Eur J Immunol,2016,46(3):665-676.
9 .张晓璇,邱海鹏,赵淑敏.丁苯酞对脑缺血再灌注大鼠HSP70及TLR4的影响研究[J].中国免疫学杂志,2014,30(10):1 401-1 403.
10 .Ma JF,Wang HM,Li QY,et al.Starvation triggers Abeta42 generation from human umbilical vascular endothelial cells[J].FEBS Lett,2010,584(14):3 101-3 106.
11 .Gambuzza ME,Sofo V,Salmeri FM,Soraci L,et al.Tolllike receptors in Alzheimer’s disease:a therapeutic perspective[J].CNS Neurol Disord Drug Targets,2014,13(9):1 542-1 558.
12 .齐凡星,胡莹,卢军栋,等.丁苯酞治疗阿尔茨海默病的临床观察[J].中国药房,2016,27(17):2 412-2 414.