CN114848842A - 以干细胞膜为靶向的纳米材料用于制备自闭症谱系障碍疾病治疗药物的应用 - Google Patents
以干细胞膜为靶向的纳米材料用于制备自闭症谱系障碍疾病治疗药物的应用 Download PDFInfo
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Abstract
本发明属纳米材料应用领域，涉及以干细胞膜为靶向的纳米材料用于制备自闭症谱系障碍疾病治疗药物的应用。其有益效果在于：提供了一种增强自闭症谱系障碍药物渗透性和靶向性的新思路。
Description
技术领域
本发明属纳米材料应用领域，涉及以干细胞膜为靶向的纳米材料用于制备自闭症谱系障碍疾病治疗药物的应用。
背景技术
自闭症谱系障碍(Autism spectrum disorder，ASD)是最常见的神经发育障碍之一，常表现为社交障碍、兴趣狭隘和刻板行为。其发病可能受环境、遗传等多种因素的影响。其中，母体免疫反应诱导的小胶质细胞激活，可能是引起后代ASD的关键因素。
相关研究表明，自闭症儿童血清中TNF-α、IL-1和IL-17都存在显着升高；且TNF-α水平与其主要症状，如社交、自我保护等呈显著正相关。该结论也为进一步研究神经炎症与ASD的关系奠定了基础(Oncotarget.2017,18,82390)
血脑屏障(BBB)是循环血液和脑组织之间的动态界面，它选择性地阻止物质进入大脑，从而提供保护性生理屏障(Leiro et al.，2017)。BBB由连续毛细管组成内皮细胞、基底膜和神经胶质膜伴有周细胞和星形胶质细胞足。相比其他组织和器官的毛细血管，脑毛细血管和相邻区域有以下三个明显的特征：(i)由脂质基组成的内皮细胞双分子膜结构与双分子膜结构紧密连接(ii)内皮细胞也被包裹连续基底膜，和(iii)星形胶质细胞存在于基底膜外，包括约大脑毛细血管表面的85％。这些结构有效防止有害物质进入脑。然而，BBB也阻断>99％的传统药物，导致治疗中枢神经系统(CNS)疾病，如神经退行性疾病、中风、脑肿瘤(Soni等人，2016)等效果差。
纳米材料因其穿越BBB的优势而吸引了神经科学领域研究人员的广泛关注(Leiro等人，2017)。一些纳米材料本身在神经精神疾病中表现出优异的生物效应，这可能取决于他们具有独特的理化性质，包括化学，电，磁和机械力(Xue等人，2014；Xue等人，2016)。纳米材料也被设计为递送治疗剂和成像剂到靶点的载体(Chang等人，2017；Godinho等人，2013；Zhao等人，2015)。诊断和治疗的结合已将诊疗一体化纳米医药作为一种重要的和潜在疾病治疗策略(Adak等人，2017；张等，2014；Urries等人，2014)。受体介导内吞作用是帮助纳米颗粒通过BBB的重要机制。
然而，在治疗脑部疾病的过程中，通过BBB输送药物仅仅是第一个挑战。常规的被动定位的靶标细胞中药物分布差和浓度低的问题也是亟需解决的另一个难点问题。而主动靶向恰好可以解决这一问题。
基于此，本发明提供了一种纳米材料，用以同时解决上述两个问题，进而用于制备治疗自闭症谱系障碍疾病药物。
发明内容
本发明所要解决的技术问题是提供一种以干细胞膜为靶向的纳米材料，能够在有效突破血脑屏障输送药物的同时具备靶向性，进而增强对于自闭症谱系障碍疾病的治疗效果。
本发明公开了一种以干细胞膜为靶向的纳米材料用于制备自闭症谱系障碍疾病治疗药物的应用，所述纳米材料包括由靶向脂质层及其包裹的介孔二氧化硅颗粒。
进一步地，所述靶向脂质层采用干细胞脂质双分子层。
进一步地，所述干细胞脂质双分子层进行了分子修饰。
优选地，所述分子修饰是将链霉亲和素通过脂质插入法修饰到干细胞脂质双分子层表面。
进一步地，所述介孔二氧化硅颗粒中浸入治疗药物。
优选地，所述治疗药物为阿司匹林。
本发明的有益效果在于：
提供了一种增强自闭症谱系障碍药物渗透性和靶向性的新思路。
附图说明
图1为实施例1中纳米材料的透射电镜图；
具体实施方式
下面结合实施例对本发明的具体实施方式作进一步描述，以下实施例仅用于更加清楚地说明本发明的技术实施例，而不能以此来限制本发明的保护范围。
实施例1
1)取小鼠干细胞，离心(1000g/min，4℃)5min，弃去上层，用1×PBS洗涤下层后，于4℃在0.25×PBS中重悬30min，离心(15000g/min，4℃)7min后制得细胞膜，细胞膜重悬并保存于双蒸水中。
2)将干细胞膜混悬液100W超声3min，得到干细胞膜囊泡。
3)将40μL链霉亲和素-PEG3400-DSPE的PBS溶液(5mg/mL)与从100μL全血中取得的干细胞膜囊泡在37℃水浴中孵育30min。
4)再与浸入阿司匹林的介孔二氧化硅纳米粒混合后使用微型挤出器挤压过100nm核孔膜，得到链霉亲和素-干细胞膜包覆的纳米材料。
对比例1
对比例1与实施例1相比，区别仅仅在于无步骤3)的修饰步骤。
对比例2
对比例1与实施例1相比，区别在于对比例2是单独使用浸入阿司匹林的介孔二氧化硅纳米粒。
为了测试本发明的有益效果，特设置如下应用例1。
应用例1
非对称结构纳米材料在自闭症谱系障碍模型鼠脑部的渗透能力测试
将制备的纳米材料配置成浓度为1mg/mL的溶液，用于制备治疗药物。为了说明其能提高在母体免疫诱导产生的自闭症幼鼠中的渗透，特做出如下检测：
首先，取三组母体免疫诱导产生的自闭症幼鼠。在其中随机选取的一组注射10μL的1mg/mL的实施例1制备的材料，记为给药组；其余分别注射对比例1及对比例2制备的材料，记为对照组1-2。
具体检测指标如下：
为在组织内观察纳米材料的的分布，采用Cy5标记纳米材料。给药4h后，用4％多聚甲醛溶液灌流，收取脑组织，并置于4％多聚甲醛溶液中浸泡过夜。之后将脑组织纵向切成厚度为1mm的脑片，小动物活体光学成像系统成像观察。可以看出：
靶向性：给药组效果最优、对照组1与给药组相当、对照组2效果最差。
透过性：给药组效果最优、对照组1次之，对照组2效果最差
由此可见，本发明公开的技术方案相比于普通纳米颗粒材料，具备更佳的脑部渗透和靶向性。
最后应说明的是：以上各实施例仅用以说明本发明的技术方案，而非对其限制；尽管参照前述各实施例对本发明进行了详细的说明，本领域的普通技术人员应当理解：其依然可以对前述各实施例所记载的技术方案进行修改，或者对其中部分或者全部技术特征进行等同替换；而这些修改或者替换，并不使相应技术方案的本质脱离本发明各实施例技术方案的范围。
Claims (6)
1.一种以干细胞膜为靶向的纳米材料用于制备自闭症谱系障碍疾病治疗药物的应用，其特征在于，所述纳米材料包括由靶向脂质层及其包裹的介孔二氧化硅颗粒。
2.根据权利要求1所述的以干细胞膜为靶向的纳米材料用于制备自闭症谱系障碍疾病治疗药物的应用，其特征在于，所述靶向脂质层采用干细胞脂质双分子层。
3.根据权利要求1所述的以干细胞膜为靶向的纳米材料用于制备自闭症谱系障碍疾病治疗药物的应用，其特征在于，所述干细胞脂质双分子层进行了分子修饰。
4.根据权利要求3所述的以干细胞膜为靶向的纳米材料用于制备自闭症谱系障碍疾病治疗药物的应用，其特征在于，所述分子修饰是将链霉亲和素通过脂质插入法修饰到干细胞脂质双分子层表面。
5.根据权利要求1所述的以干细胞膜为靶向的纳米材料用于制备自闭症谱系障碍疾病治疗药物的应用，其特征在于，所述介孔二氧化硅颗粒中浸入治疗药物。
6.根据权利要求1所述的以干细胞膜为靶向的纳米材料用于制备自闭症谱系障碍疾病治疗药物的应用，其特征在于，所述治疗药物为阿司匹林。
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