Safety, efficacy, and molecular mechanism of claudin-1-specific peptides to enhance blood-nerve-barrier permeability.
Safety, efficacy, and molecular mechanism of claudin-1-specific peptides to enhance blood-nerve-barrier permeability.
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DOI：
10.1016/j.jconrel.2014.04.029
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发表时间：
2014-07
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通讯作者：
Reine-Solange Sauer;S. Krug;D. Hackel;Christian Staat;Natalia Konasin;Shaobing Yang;Benedikt Niedermirtl;Judith Bosten;Ramona Günther;S. Dąbrowski;K. Doppler;C. Sommer;I. Blasig;A. Brack;H. Rittner
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作者：
Reine-Solange Sauer;S. Krug;D. Hackel;Christian Staat;Natalia Konasin;Shaobing Yang;Benedikt Niedermirtl;Judith Bosten;Ramona Günther;S. Dąbrowski;K. Doppler;C. Sommer;I. Blasig;A. Brack;H. Rittner
血神经屏障由神经束膜和神经内膜血管组成。神经束膜屏障由基底膜和一层神经束膜细胞组成，神经束膜细胞被紧密连接蛋白封闭，防止例如用于选择性局部疼痛控制的镇痛剂的应用。血神经屏障中的屏障密封蛋白之一是claudin-1。因此，开发了来自claudin-1上的第一胞外环（ECL 1）的claudin-1-肽模拟物（C1 C2）。在这项研究中，我们进一步评估了紧密连接蛋白在Wistar大鼠神经束膜中的表达，并表征了C1 C2的特异性、体内适用性、作用机制以及生物相容性。在神经束膜中，claudin-19、tricellulin和ZO-1表达，而claudin-2、3、8和-11不表达。C1 C2特异性结合于紧密连接蛋白-1和荧光5，6-羧基四甲基罗丹明-C1 C2的ECL 1被迅速内化。用C1 C2打开神经束膜降低了claudin-1的mRNA和蛋白表达，并增加了小分子和大分子渗透到周围神经中。C1 C2的应用促进了使用μ-阿片受体激动剂如DAMGO或吗啡的区域镇痛，而没有运动损伤的幼稚大鼠以及具有后爪炎症的大鼠。与此相反，来自于claudin-2上的ECL 1的对照肽C2 C2既不打开屏障也不促进阿片类药物介导的区域镇痛。C1 C2输送耐受性良好，未造成形态和功能性神经损伤。C1 C2的影响可以逆转干扰wnt信号转导途径，特别是同源框转录因子cdx 2，使用糖原合成酶激酶-3抑制剂。总之，我们描述的组成和一个途径，以打开神经束膜屏障，采用肽传递亲水性物质到周围神经。
The blood–nerve barrier consists of the perineurium and endoneurial vessels. The perineurial barrier is composed of a basal membrane and a layer of perineurial cells sealed by tight junction proteins preventinge.g.application of analgesics for selective regional pain control. One of the barrier-sealing proteins in the blood–nerve barrier is claudin-1. Therefore, the claudin-1-peptidomimetics (C1C2), derived from the first extracellular loop (ECL1) on claudin-1 was developed. In this study, we further evaluated the expression of tight junction proteins in the perineurium in Wistar rats and characterized the specificity,in vivoapplicability, mechanism of action as well as the biocompatibility of C1C2. In the perineurium, claudin-19, tricellulin and ZO-1, but no claudin-2, 3, 8 and -11 were expressed. C1C2 specifically bound to the ECL1 of claudin-1 and fluorescent 5,6-carboxytetramethylrhodamine-C1C2 was rapidly internalized. Opening the perineurium with C1C2 reduced the mRNA and protein expression of claudin-1 and increased small and macromolecule permeability into the peripheral nerve. Application of C1C2 facilitated regional analgesia using μ-opioid receptor agonists like DAMGO or morphine without motor impairment in naïve rats as well as rats with hind paw inflammation. In contrast the control peptide C2C2 derived from ECL1 on claudin-2 did neither open the barrier nor facilitated opioid-mediated regional analgesia. C1C2 delivery was well tolerated and caused no morphological and functional nerve damage. C1C2 effects could be reversed by interference with the wnt-signal-transduction pathway, specifically the homeobox transcription factor cdx2, using a glycogen-synthase-kinase-3 inhibitor. In summary, we describe the composition of and a pathway to open the perineurial barrier employing a peptide to deliver hydrophilic substances to the peripheral nerve.