Tight Junction Modulatory Fusion Peptide (ADT-6) Enhances GFP Protein Permeability through the Paracellular Pathway in Caco-2 Cell Lines: An In-Vitro Study.

IF 1.2 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY
Salimeh Hassani, Keyvan Nedaei, Rahim Jafari, Ghasem Bagherpour
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引用次数: 0

Abstract

Background: The oral delivery of therapeutic peptides and proteins presents a significant challenge in pharmaceutical development due to barriers such as the intestinal epithelium and the blood-brain barrier (BBB). These barriers limit the passage of large, hydrophilic molecules through transcellular pathways and restrict paracellular transport due to intercellular tight junctions. This study investigates the potential of E- cadherin-modulating peptide, ADT-6, to improve the penetration of these therapeutic agents.

Methods: We constructed a fusion protein of ADT-6 and green fluorescent protein (GFP) to evaluate its activity and transport through the epithelial cells' paracellular pathway. Using Escherichia coli strains for expression, we cloned the GFP-ADT-6 construct, which provides a solid foundation for our study's methodology.

Results: Our molecular simulations showed that the linker between GFP and ADT-6 maintains the fusion protein's integrity and provides flexibility in receptor interaction. Permeability experiments revealed that ADT-6 markedly reduced transepithelial electrical resistance (TEER) and significantly increased GFP transfection in Caco-2 cell monolayers dose-dependently. Results of ELISA confirmed these findings, showing high GFP levels in the lower compartment of Transwell systems treated with GFP-ADT-6.

Conclusions: This study demonstrates the potential of ADT-6 to deliver proteins from the paracellular route, enhance the bioavailability of pharmaceutical drugs by altering cell-cell interactions, and provide new opportunities for oral drug delivery strategies.

紧密连接调节融合肽(ADT-6)通过Caco-2细胞系的细胞旁通路增强GFP蛋白的通透性:一项体外研究
背景:由于肠上皮和血脑屏障(BBB)等屏障的存在,治疗性肽和蛋白质的口服递送在药物开发中面临着重大挑战。这些屏障限制了大的亲水分子通过跨细胞途径,并由于细胞间紧密连接而限制了细胞旁运输。本研究探讨了E-钙粘蛋白调节肽ADT-6的潜力,以提高这些治疗剂的渗透。方法:构建ADT-6与绿色荧光蛋白(GFP)的融合蛋白,考察其活性及其在上皮细胞旁通路的转运。利用大肠杆菌菌株进行表达,克隆了GFP-ADT-6构建体,为本研究的方法奠定了坚实的基础。结果:我们的分子模拟表明,GFP和ADT-6之间的连接物保持了融合蛋白的完整性,并提供了受体相互作用的灵活性。渗透性实验显示,ADT-6显著降低Caco-2细胞单层的上皮电阻值(TEER),并显著增加GFP转染量,呈剂量依赖性。ELISA结果证实了这些发现,显示GFP- adt -6处理的Transwell系统的下隔室中GFP水平较高。结论:本研究证明了ADT-6从细胞旁途径递送蛋白质的潜力,通过改变细胞间相互作用提高药物的生物利用度,并为口服药物递送策略提供了新的机会。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Reports of Biochemistry and Molecular Biology
Reports of Biochemistry and Molecular Biology BIOCHEMISTRY & MOLECULAR BIOLOGY-
CiteScore
2.80
自引率
23.50%
发文量
60
审稿时长
10 weeks
期刊介绍: The Reports of Biochemistry & Molecular Biology (RBMB) is the official journal of the Varastegan Institute for Medical Sciences and is dedicated to furthering international exchange of medical and biomedical science experience and opinion and a platform for worldwide dissemination. The RBMB is a medical journal that gives special emphasis to biochemical research and molecular biology studies. The Journal invites original and review articles, short communications, reports on experiments and clinical cases, and case reports containing new insights into any aspect of biochemistry and molecular biology that are not published or being considered for publication elsewhere. Publications are accepted in the form of reports of original research, brief communications, case reports, structured reviews, editorials, commentaries, views and perspectives, letters to authors, book reviews, resources, news, and event agenda.
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