An intestinal paracellular pathway biased toward positively-charged macromolecules.
An intestinal paracellular pathway biased toward positively-charged macromolecules.
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DOI：
10.1016/j.jconrel.2018.09.003
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发表时间：
2018-10-28
期刊：
影响因子：
--
通讯作者：
Mrsny RJ
中科院分区：
文献类型：
--
作者：
Almansour K;Taverner A;Turner JR;Eggleston IM;Mrsny RJ
由于缺乏一种有效的机制来安全和持续地提高肠道上皮细胞对大分子的摄取，成功开发口服治疗多肽药物的前景仍然不太可能。我们以前通过确定一种内源性机制来控制肠道细胞旁通透性，这种机制可以被一种名为PIP 640的多肽激活，这种多肽可以增加S19位的磷酸化肌球蛋白轻链(MLC-PS19)的细胞水平。在体外或体内根尖应用表明，在几分钟内增加大分子溶质的传输，在移除后几个小时内完全恢复。我们现在研究PIP 640介导的通透性变化的性质。经PIP 640处理的融合Caco-2细胞单层可促进4 kDa的右旋糖苷转运，但不能促进10 kDa的右旋糖苷转运。紧密连接(TJ)蛋白的表达和/或细胞分布的变化仅限于Claudin-2在一段时间内的增加，这与其分布从细胞核向细胞膜部分的明显转移有关。PIP640介导的上皮改变有别于促炎细胞因子肿瘤坏死因子-α和干扰素-γ的联合作用。同时，肿瘤坏死因子-α/干扰素-γ治疗也增加了MLC-PS19的水平，这些细胞因子促进了AB对70-kDa右旋糖苷的转运，并降低了TJ的阻滞素表达。PIP 640诱导的Claudin-2依赖的变化导致了正电荷大分子的AB转运偏向，使用4-kDa葡聚糖的电荷变体和通过比较鲑鱼降钙素和埃塞那肽的转运在体外证明了这一点。在大鼠空肠腔内注射后，体内显示了类似的结果，即Claudin-2在TJ的定位增加，这些偏向阳离子特性的治疗性多肽的转运增加。综上所述，这些数据显示了PIP 640在小的治疗性多肽的大小范围内增强溶质的细胞旁渗透性的潜在机制，该小的治疗性多肽偏向于带正电荷的溶质。
Lacking an effective mechanism to safely and consistently enhance macromolecular uptake across the intestinal epithelium, prospects for successful development of oral therapeutic peptide drugs remain unlikely. We previously addressed this challenge by identifying an endogenous mechanism that controls intestinal paracellular permeability that can be activated by a peptide, termed PIP 640, which can increase cellular levels of phosphorylated myosin light chain at position S19 (MLC-pS19). Apical application in vitro or luminal application in vivo was shown to increase macromolecular solute transport within minutes that recovered completely within a few hours after removal. We now examine the nature of PIP 640-mediated permeability changes. Confluent Caco-2 cell monolayers treated with PIP 640 enhanced apicalto-basolateral (AB) transport of 4-kDa, but not 10-kDa, dextran. Expression and/or cellular distribution changes of tight junction (TJ) proteins were restricted to increased claudin-2 over a time course that correlated with an apparent shift in its distribution from the nucleus to the membrane fraction of the cell. PIP 640-mediated epithelial changes were distinct from the combined actions of the pro-inflammatory cytokines tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ). While TNF-α/IFN-γ treatment also increased MLC-pS19 levels, these cytokines enhanced AB transport for 70-kDa dextran and decreased occludin expression at TJs. Claudin-2-dependent changes induced by PIP 640 resulted in an AB transport bias for positively-charged macromolecules demonstrated in vitro using charge variants of 4-kDa dextrans and by comparing transport of salmon calcitonin to exenatide. Comparable outcomes of increased TJ localization of claudin-2 and enhanced transport of these therapeutic peptides that biased toward cationic characteristics was demonstrated in vivo following after intraluminal injection into rat jejunum. Together, these data have shown a potential mechanism for PIP 640 to enhance paracellular permeability of solutes in the size range of small therapeutic peptides that is biased toward positively-charged solutes.
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影响因子：
7.8
作者：
Furuse, Mikio;Hata, Masaki;Furuse, Kyoko;Yoshida, Yoko;Haratake, Akinori;Sugitani, Yoshinobu;Noda, Tetsuo;Kubo, Akiharu;Tsukita, Shoichiro
通讯作者：
Tsukita, Shoichiro
影响因子：
14
作者：
Krug, Susanne M.;Amasheh, Maren;Amasheh, Salah
通讯作者：
Amasheh, Salah
DOI：
10.1073/pnas.91.5.1942
发表时间：
1994-03-01
影响因子：
11.1
作者：
PAPPENHEIMER, JR;DAHL, CE;MAGGIO, JE
通讯作者：
MAGGIO, JE
影响因子：
7.3
作者：
Lingaraju A;Long TM;Wang Y;Austin JR 2nd;Turner JR
通讯作者：
Turner JR
影响因子：
3.1
作者：
Lu Z;Ding L;Lu Q;Chen YH
通讯作者：
Chen YH