Attenuation of Kindled Seizures by Intranasal Delivery of Neuropeptide-Loaded Nanoparticles
Attenuation of Kindled Seizures by Intranasal Delivery of Neuropeptide-Loaded Nanoparticles
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DOI：
10.1016/j.nurt.2009.02.001
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发表时间：
2009-04-01
影响因子：
5.7
通讯作者：
Veronesi, Michael C.
中科院分区：
文献类型：
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作者：
Kubek, Michael J.;Domb, Abraham J.;Veronesi, Michael C.
促甲状腺激素释放激素（TRH; Protirelin）是一种内源性神经肽，已知在动物癫痫模型和某些难治性癫痫患者中具有抗惊厥作用。然而，其作用的持续时间受到快速组织代谢和血脑屏障的限制。神经肽在缓释生物可降解纳米颗粒（NPs）中的直接鼻-脑递送是提高CNS神经肽生物利用度的一种有前途的治疗模式。为了提供这种递送方法的原理证据，我们使用颞叶癫痫点燃模型来表明：1）在癫痫灶中植入载有TRH的共聚物微盘可以在行为阶段、后放电持续时间（ADD）和阵挛持续时间方面减弱点燃发展; 2）鼻内给予未保护的TRH类似物可以在一定浓度下急性抑制完全点燃的癫痫发作，依赖的方式在ADD和癫痫发作阶段;和3）鼻内施用含有TRH（TRH-NPs）的聚乳酸纳米颗粒（PLA-NPs）可以在行为阶段、ADD和阵挛持续时间方面阻碍点燃发展。此外，我们在大鼠中鼻内递送负载荧光染料的PLA-NP，并将负载染料或附着染料的NP应用于培养的皮质神经元，以分别证明NP在体内和体外随时间的吸收和分布。此外，纳米颗粒免疫染色方法被开发为直接可视化神经肽负载的纳米颗粒的组织水平和分布的程序。总的来说，这些数据为TRH-NP的鼻内递送提供了概念证明，作为1）抑制癫痫发作和可能的癫痫发生和2）成为鼻内抗惊厥纳米颗粒治疗剂的先导化合物的可行手段。
Thyrotropin-releasing hormone (TRH; Protirelin), an endogenous neuropeptide, is known to have anticonvulsant effects in animal seizure models and certain intractable epileptic patients. Its duration of action, however, is limited by rapid tissue metabolism and the blood-brain barrier. Direct nose-to-brain delivery of neuropeptides in sustained-release biodegradable nanoparticles (NPs) is a promising mode of therapy for enhancing CNS neuropeptide bioavailability. To provide proof of principle for this delivery approach, we used the kindling model of temporal lobe epilepsy to show that 1) TRH-loaded copolymer microdisks implanted in a seizure focus can attenuate kindling development in terms of behavioral stage, after-discharge duration (ADD), and clonus duration; 2) intranasal administration of an unprotected TRH analog can acutely suppress fully kindled seizures in a concentration-dependent manner in terms of ADD and seizure stage; and 3) intranasal administration of polylactide nanoparticles (PLA-NPs) containing TRH (TRH-NPs) can impede kindling development in terms of behavioral stage, ADD, and clonus duration. Additionally, we used intranasal delivery of fluorescent dye-loaded PLA-NPs in rats and application of dye-loaded or dye-attached NPs to cortical neurons in culture to demonstrate NP uptake and distribution over time in vivo and in vitro respectively. Also, a nanoparticle immunostaining method was developed as a procedure for directly visualizing the tissue level and distribution of neuropeptide-loaded nanoparticles. Collectively, the data provide proof of concept for intranasal delivery of TRH-NPs as a viable means to 1) suppress seizures and perhaps epileptogenesis and 2) become the lead compound for intranasal anticonvulsant nanoparticle therapeutics.