摘要
目的:研究丁苯酞对β-淀粉样蛋白1-42(Aβ_(1-42))诱导人脐静脉内皮细胞(HUVEC)凋亡的保护作用及其机制。方法:将HUVEC分为正常对照组、Aβ_(1-42)组、TAK242组(10 nmol/L)、二甲基亚砜(DMSO)组(1‰DMSO)和丁苯酞低、中、高浓度组(40、80、160μg/L),除正常对照组和DMSO组外,其余各组细胞均加入50μmol/L Aβ1_(1-42)培养HUVEC 24 h,同时TAK242组、DMSO组和丁苯酞低、中、高浓度组细胞还加入相应浓度的药物作用30 min,每个浓度3个复孔。CCK-8法测定细胞活力,Hochest 33342/PI双染法观察细胞凋亡情况,膜联蛋白(Annexin)Ⅴ-异硫氰酸荧光素(FITC)流式细胞仪检测细胞凋亡率,Western blot法检测细胞中果蝇样受体4(TLR4)、环氧合酶2(COX-2)蛋白表达,ELISA法检测细胞中白细胞介素1(IL-1)、肿瘤坏死因子α(TNF-α)含量。结果:与正常对照组比较,Aβ_(1-42)组细胞活力减少、凋亡率增加,TLR4、COX-2蛋白表达和IL-1、TNF-α含量增加;与Aβ_(1-42)组比较,TAK242组和丁苯酞低、中、高浓度组细胞活力增加、凋亡率减少,TLR4、COX-2蛋白表达和IL-1、TNF-α含量减少,差异均有统计学意义(P<0.05或P<0.01)。结论:丁苯酞可改善Aβ_(1-42)所致的HUVEC凋亡,其机制可能与抑制TLR4、COX-2和炎症因子表达有关。
OBJECTIVE： To study the protective effect of butylphthalide on the apoptosis of human umbilical vein endothelial cells （HUVECs） induced by Aβ1-42 and its mechanism. METHODS： HUVECs were divided into normal control group, A I3 t-42 group, TAK242 group （10 nmol/L）, DMSO group （1‰DMSO） and butylphthalide low-concentration, medium-concentration and high-concentration groups （40, 80, 160 μg/L）. Except for normal control group and DMSO group, other groups were given 50 μmol/L Aβ1-42 to culture HUVECs for 24 h. TAK242 group, DMSO group and butylphthalide low-concentration, medium-concentra- tion and high-concentration groups were given relevant concentration of drugs for 30 min, with 3 holes for each concentration. The cell viability was determined by CCK-8 assay; cell apoptosis was observed by Hochest 33342/PI double staining; the cell apoptotic rate was detected by Annexin V-fluorescein isothiocyanate （FITC） flow cytometry; the protein expression of TLR-4 and COX-2 were determined by Western blot assay; the contents of IL-1 and TNF-a were detected by ELISA. RESULTS： Compared with nor- mal control group, cell viability of HUVECs were decreased in A13~_42 group; while apoptotic rate, protein expression of TLR4 and COX-2, the contents of IL-1 and TNF-ct were increased. Compared with Aβ1-42 group, cell viability of HUVECs were increased in TAK242 group and butylphthalide low-concentration, medium-concentration and high-concentration groups; while apoptotic rate, protein expression of TLR4 and COX-2, the contents of IL-1 and TNF-a were decreased, with statistical significance （P〈0.05 or P〈0.01 ）. CONCLUSIONS： Butylphthalide can reduce HUVECs apoptosis induced by Aβ1-42, which may be related with inhibiting the expression of TLR4, COX-2 and inflammatory factors.
作者
黄江
姜鲜
宋大强
刘明华
章卓
HUANG Jiang JIANG Xian SONG Daqiang LIU Minghua ZHANG Zhuol(Pharmacology Teaching and Research Section, School of Pharmacy, Southwest Medical University, Sichuan Luzhou 646000, China Dept. of Pharmacy, the Affiliated TCM Hospital of Southwest Medical University, Sichuan Luzhou 646000, China Dept. of Anesthesiology, the Affiliated Hospital of Southwest Medical University, Sichuan Luzhou 646000, China)
出处
《中国药房》
CAS
北大核心
2017年第4期483-486,共4页
China Pharmacy
基金
四川省科技厅一般项目(No.2014S20071)
泸州市科技局科技支撑计划项目(No.2013LZLY-J52)
