Intracellular Angiotensin-II Measurement in Streptozotocin-Induced Rat Vascular Smooth Muscle Cells and Its Relationship with Angiotensin-II Receptors

Year 2026, Issue: Advanced Online Publication, 1 - 15

Zehra Çiçek , Kübra Akıllıoğlu , Zehra Gül Yaşar , Ayşe Doğan

https://doi.org/10.18621/eurj.1760073

Abstract

Objectives: Angiotensin II (Ang-II) is vital constituent of renin angiotensin aldosterone system and increases in some cardiovascular diseases such as diabetes. However, there are not enough studies related to intracellular and extracellular Ang-II levels and its interaction with Ang-II type 1 and 2 receptors (ATR1, ATR2) in vascular smooth muscle cells (VSMCs). We aimed to investigate healthy and diabetic rat model of VSMCs (H-VSMCs, D-VSMCs) proliferation and Ang-II levels.

Methods: VSMCs were isolated from the aorta of healthy and diabetic Wistar rats. Diabetic model was achieved by intravenous administration of 45 mg/kg streptozotocin. Firstly, different Ang-II (0-1000 μM) concentration was performed for dose study. Ang-II (0.1 μM), Ang-II type 1 receptor (ATR1) antagonist (Olmesartan, 1 μM) and Ang-II type 2 receptor (ATR2) antagonist (PD123,319, 1 μM) were practiced together, and thereafter cell proliferation was evaluated by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) method. Intracellular and extracellular Ang-II levels were measured by ELISA kit.

Results: While H-VSMCs proliferation increased in Ang-II 0.1, 0.01, 0.001 and 0.0001 μM, D-VSMCs proliferation increased Ang-II 0.1 and 0.01 μM applications (P<0.05). Olmesartan 1 µM inhibited proliferation in H-VSMCs. Ang-II detected intracellular and extracellular groups of VSMCs, but no significant difference was found between H-VSMCs and D-VSMCs groups (P˃0.05).

Conclusions: Ang-II enhances proliferation of H-VSMCs and D-VSMCs. There is no relationship that could be established between intracellular and extracellular Ang-II levels, H-VSMCs and D-VSMCs proliferation and Ang-II receptors.

Keywords

Angiotensin II , Diabetes , Smooth Muscle Cell , Vascular

Ethical Statement

The present study was approved by Çukurova University Animal Experiments Local Ethics Committee (Decision No: 2016/11-1 and date: 22.12.2016, Adana, Türkiye). All experimental procedures were carried out accordance with the U.K. Animals (Scientific Procedures) Act, 1986 and associated guidelines, EU Directive 2010/63/EU for animal experiments, or the National Institutes of Health guide for the care and use of Laboratory animals (NIH Publications No. 8023, revised 1978).

Supporting Institution

This study was funded by the Çukurova University with a scientific research project (Adana, Türkiye)

Project Number

CÜBAP-TTU No. 2017-8071

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