纳米化杀手肽的制备表征及抗新生隐球菌感染的实验研究
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上海市科委纳米专项基金(05nm50010).


Killer peptide-loaded nanoparticles: characterization and evaluation of anticryptcoccal efficacy
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Supported by the Nanotechnology Foundation of Shanghai Science and Technology Committee(05nm50010).

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    摘要:

    目的:以聚乳酸-羟基乙酸(PLGA)作为杀手肽(killer peptide,KP)的有效载体,制备出对隐球菌感染疗效更佳的载药纳米粒。方法:采用复乳法制备杀手肽PLGA纳米粒,并使用透射电镜、扫描电镜、动态光散射、测定包封率、考察体外释放等手段对其表征;采用平板计数法验证杀手肽和杀手肽PLGA纳米粒对新生隐球菌ATCC32609的体外杀灭作用;构建小鼠隐球菌系统感染模型,通过生存期和重要脏器染菌量的变化比较纳米制剂同游离杀手肽的疗效变化。结果:成功制备出杀手肽PLGA纳米粒。人工合成杀手肽和杀手肽PLGA纳米粒均具有较强体外抗新生隐球菌ATCC32609作用,与生理盐水组比较有统计学差异(P<0.05),但杀手肽PLGA纳米粒与游离杀手肽之间比较无统计学差异(P>0.05)。动物实验中,同游离杀手肽比较,纳米制剂使隐球菌系统感染小鼠生存期明显延长(P<0.05),重要脏器染菌量显著下降:纳米化杀手肽 3 mg/kg和5 mg/kg剂量组同等剂量游离杀手肽组比较,差异有统计学意义(P<0.05),但空白PLGA纳米粒同生理盐水组比较差异无统计学意义(P>0.05)。结论:纳米材料包裹可显著增强抗系统性隐球菌感染疗效。

    Abstract:

    Objective:To prepare killer peptide-loaded PLGA nanoparticles which have better effect in treating systemic crypotococcosis.Methods: We adopted a modified double-emulsion method to prepare the killer peptide-loaded PLGA nanoparticles,and the nanoparticles were characterized by using TEM,SEM,Dynamic Laser Scattering,drug loading analysis,ecapsulation efficacy assessment and in vitro release test.We also evaluated the antifungal activities of killer peptide and killer peptide-loaded PLGA nanoparticles against Cryptococcus neoformans ATCC32609 by plate count method.In the in vivo study,the anticryptococcal efficacies of killer peptide and killer peptide-loaded PLGA nanoparticles were compared by observing the survival and colony-forming in vital organs in a systemic murine cryptococcosis model.Results: We successfully synthesized killer peptide-loaded PLGA nanoparticles.In vitro anticryptococcal test showed that the killer peptide and killer peptide-loaded PLGA nanoparticles had significantly stronger anticryptococcal effect compared with the normal saline group (P<0.05),and there was no significant difference in the efficacies between free killer peptide and killer peptide-loaded PLGA nanoparticles (P>0.05).In vivo test demonstrated that mice treated with killer peptide-loaded PLGA nanoparticles (3 mg/kg or 5 mg/kg) had longer survival period and less fungal burden in vital organs than mice treated with free killer peptide (P<0.05); there was no significant difference in fungal burden and survival period between empty PLGA nanoparticle treated group and normal saline group (P>0.05).Conclusion: The killer peptide-loaded PLGA nanoparticles are more effective in treating systemic murine cryptococcosis.

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  • 收稿日期:2008-06-18
  • 最后修改日期:2009-03-08
  • 录用日期:2009-03-11
  • 在线发布日期: 2009-05-22
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