摘要
目的:制备负载2-甲基-α-卡波林-3-甲酰胺(2MAC3C)的两亲性壳聚糖纳米胶束,以改善其水溶性,并研究该载药胶束对大鼠心肌细胞(H9c2)缺氧/复氧(H/R)损伤活性的影响。方法:以天然高分子材料壳聚糖为骨架,衍生化为羟乙基壳聚糖-辛酸偶联物(OHC);采用~1H-NMR、红外分光光度法确证OHC的结构;透析法制备2MAC3C-OHC纳米胶束;芘荧光光谱法测定纳米胶束的临界胶束浓度(CMC);透射电镜观察纳米胶束形态;粒度测定仪考察其粒径及电位;离心法考察制剂的包封率及载药量;运用四甲基偶氮唑盐(MTT)法考察2MAC3C-OHC对H9c2心肌细胞毒性及H/R损伤活性的影响。结果:2MAC3C-OHC的CMC为0.250 g/L;形态为球形;平均粒径为(227.7±3.0)nm;Zeta电位为(9.84±0.72)mV;包封率为(68.00±3.67)%;载药量为(19.32±0.11)%;与2MAC3C溶液组相比,2MAC3C-OHC对H9c2心肌细胞的毒性减弱;2MAC3C-OHC能显著提高损伤后H9c2心肌细胞存活率(P<0.001)。结论:所制备的2MAC3C-OHC粒径小,载药量高,提高了2MAC3C对H/R损伤后H9c2心肌细胞存活率,同时改善了较高浓度时2MAC3C对H9c2心肌细胞毒性。研究结果对2MAC3C-OHC在缺氧/复氧(H/R)损伤或心肌缺血再灌注损伤(MIRI)治疗中的应用奠定了基础。
Objective:To prepare amphiphilic chitosan nanoparticles loaded with 2-methyl-α-carboline-3-carboxamide(2 MAC3 C)to improve the water solubility,and to study the effect of the drug-loaded micelles on hypoxia/reoxygenation(H/R)injury activity of rat cardiac myocytes(H9 c2).Methods:The hydroxyethyl chitosan-octanoic acid conjugate polymer(OHC)was prepared with the natural polymer material chitosan as the steleton and its structure was determined by ~1H-NMR and IR.2 MAC3 C-OHC nano-micelle was prepared by the dialysis method and the critical micelle concentration(CMC)of nano-micelle was determined by Pyrene fluorescence spectrometry.The transmission electron microscope(TEM)was used to observe the nano-micelle morphology,and the particle size tester was used to detect the diameter and Zeta potential.Ultracentrifugation was utilized to determine the encapsulation rate and drug-loading rate.The effect of 2 MAC3 C-OHC on the cytotoxicity of H9 c2 myocardial cells and H/R injury activity was investigated by MTT.Results:The CMC of 2 MAC3 C-OHC was 0.250 g/L,the shape was spherical with a nano-metric size of(227.7±3.0)nm and the Zeta potential was(9.84±0.72)mV.The encapsulation rate was(68.00±3.67)%,and drug-loading rate was(19.32±0.11)%.Compared with 2 MAC3 C solution group,2 MAC3 C-OHC showed lower cytotoxicity on H9 c2 cardiomyocytes,and significantly increased the survival rate of H9 c2 cardiomyocytes after injured(P<0.001).Conclusion:The prepared 2 MAC3 C-OHC has a small particle size and a high drug-loading rate,which ultimately improves the survival rate of H9 c2 cardiomyocytes against H/R injury,and decreases the toxicity of 2 MAC3 C to H9 c2 cardiomyocytes at higher concentrations.The results lay a foundation for the application of 2 MAC3 C-OHC in the treatment of H/R injury or myocardial ischemia reperfusion injury(MIRI).
作者
陈婷
王磊
周孟
张金娟
梁冰
廖尚高
CHEN Ting;WANG Lei;ZHOU Meng;ZHANG Jin-juan;LIANG Bing;LIAO Shang-gao(State Key Laboratory of Functions and Applications of Medicinal Plants/School of Pharmacy,Guizhou Medical University,Gui'an New District 550025,China;Engineering Research Center for the Development and Application of Ethnic Medicine and TCM(Ministry of Education),Guizhou Medical University,Guiyang 550004,China;School of Basic Medical Sciences,Guizhou Medical University,Gui'an New District 550025,China)
出处
《中药材》
CAS
北大核心
2019年第5期1107-1111,共5页
Journal of Chinese Medicinal Materials
基金
贵州医科大学博士启动基金(院博合j字2017-022)
国家自然科学基金(81360635,81660570)
贵州省科技厅基础研究计划(黔科合基础[2017]1156)
大学生创新创业项目(2018520356).
