Selection of a high-affinity and in vivo bioactive ssDNA aptamer against angiotensin II peptide.
Unique features of aptamers have attracted interests for a broad range of applications. Aptamers are able to specifically bind to targets and inhibit their functions. This study, aimed to isolate the high affinity ssDNA aptamers against bio-regulator peptide angiotensin II (Ang II) and investigate their bioactivity in cellular and animal models. To isolate ssDNA aptamers, 12 rounds of affinity chromatography SELEX (Systematic Evolution of Ligands by EXponential enrichment) procedure were carried out. The SPR (surface plasmon resonance) and ELONA (enzyme linked oligonucleotide assay) analysis were used to determine the affinity and specificity of aptamers. The ability of selected aptamers to inhibit the proliferative effect of Ang II on human aortic vascular smooth muscle cells (HA-VSMCs) and their performance on Wistar rat urinary system and serum electrolyte levels were investigated. Two full-length aptamers (FLC112 and FLC125) with high affinity of respectively 7.52±2.44E-10 and 5.87±1.3E-9M were isolated against Ang II. The core regions of these aptamers (CRC112 and CRC125) also showed affinity of 5.33±1.15E-9 and 4.11±1.09E-9M. In vitro analysis revealed that FLC112 and FLC125 can inhibit the proliferative effect of Ang II on HA-VSMCs (P<0.05). They also significantly reduced the serum sodium level and increased the urine volume (P<0.05). The core regions of aptamers did not show high inhibitory potential against Ang II. It can be a spotlight that ssDNA aptamers have high potential for blocking Ang II. In conclusion, it appears that the researches focusing on high affinity and bioactive aptamers may lead to excellent results in blocking Ang II activity.
Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Mohammad
M
Heiat
Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran. Electronic address: ranjbarre@gmail.com.
Reza
R
Ranjbar
Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Ali Mohammad
AM
Latifi
Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Mohammad Javad
MJ
Rasaee
D000804:Angiotensin II / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D002460:Cell Line; D002846:Chromatography, Affinity; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D008024:Ligands; D009131:Muscle, Smooth, Vascular / Q000187:drug effects; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D051381:Rats; D052156:SELEX Aptamer Technique; D020349:Surface Plasmon Resonance
D052157:Aptamers, Nucleotide; D004277:DNA, Single-Stranded; D008024:Ligands; D010455:Peptides; D000804:Angiotensin II
Angiotensin II; Aptamer; SELEX; Serum electrolytes; Urine volume; VSMCs