Construction of hydrogen peroxide-scavenging liposomes based on vitamin E and evaluation of its in vitro anticoagulant effect
10.16016/j.2097-0927.202404044
- VernacularTitle:水蛭素-草酸二维生素E酯脂质体的构建及其体外抗凝作用评价
- Author:
Qinghua YANG
1
;
Jianxiang ZHANG
;
Zhenhua ZHOU
Author Information
1. 400038 重庆,陆军军医大学(第三军医大学)第一附属医院神经内科
- Keywords:
hydrogen peroxide scavenging;
liposome;
anti-inflammatory and antioxidant;
anticoagulant effect;
oxalate vitamin E;
hirudin
- From:
Journal of Army Medical University
2024;46(21):2475-2484
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct vitamin E(tocopherol,VE)-based liposomes with both hydrogen peroxide-scavenging function and anticoagulant effect,and explore whether they can be used to treat acute ischemic stroke and then evaluate their in vitro anticoagulant effect.Methods Two molecules of VE were chemically bonded through peroxalate bond to obtain oxalate vitamin E(OVE).The anticoagulant unit LA-Hirudin was obtained by amidation reaction by connecting the anticoagulant drug hirudin to linoleic acid(LA).The structures of the synthesized products were characterized by using H nuclear magnetic resonance(1HNMR)and FT-IR spectra.Hirudin-Oxalic acid divitamin E ester liposome(HOL)was prepared by thin film dispersion and characterized with dynamic light scattering(DLS)and transmission electron microscopy(TEM).Mouse brain-derived endothelial cells(bEnd3)and human umbilical vein endothelial cells(HUVEC)were applied as the object of cell experiments.The levels of H2O2,and cellular inflammatory factors TNF-α,IL-6 and IL-1 β were determined by ELISA to evaluate the effect of the liposomes on the severity of cellular inflammation.Cell counting kit-8(CCK-8)assay and cell proliferation assay(5-Ethynyl-2'-deoxyuridine,Edu)were used to detect the effects of the liposomes on cell count and proliferation.The anticoagulant effect of the prepared liposomes was detected by using whole blood of mice.Results 1HNMR spectrum showed that OVE was successfully synthesized,and the FT-IR spectrum displayed that hirudin and linoleic acid were successfully bonded to obtain LA-Hirudin.The results of DLS and TEM indicated that the prepared liposomes were spheroids,in uniform distribution,with an average particle size of 161 nm.The cell experiments showed that the liposomes significantly inhibited the secretion of H2O2 and inflammatory factors,and exerted obvious antioxidant and anti-inflammatory effects in the model cells(P＜0.05).CCK-8 assay and Edu proliferation assay proved that the liposomes suppressed the decreased cell count and alleviated the inhibited cell proliferation caused by LPS(P＜0.01).Anticoagulation experiment revealed that the liposomes had significant inhibitory effect of CaCl2-induced coagulation reaction(P＜0.01).Conclusion A hydrogen peroxide-scavenging function material,OVE is synthesized,and then anticoagulant drug hirudin and OVE are used to prepare OVE thrombolytic liposomes,HOL.The liposomes have antioxidant and anti-inflammatory effects at the cellular level and also have significant anticoagulant effect.
