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

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

doi:10.18621/eurj.1760073

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

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

Supporting Institution

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

Project Number

CÜBAP-TTU No. 2017-8071

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).

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Details

Primary Language

English

Subjects

Cell Development, Proliferation and Death , Cell Physiology

Journal Section

Research Article

Authors

Zehra Çiçek ^*
0000-0003-3205-5463
Türkiye

Kübra Akıllıoğlu
0000-0003-4896-8822
Türkiye

Zehra Gül Yaşar
0000-0001-6660-2643
Türkiye

Ayşe Doğan
0000-0002-6508-5266
Türkiye

Early Pub Date

October 21, 2025

Publication Date

-

Submission Date

August 7, 2025

Acceptance Date

October 6, 2025

Published in Issue

Year 2026 Number: Advanced Online Publication

DOI

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

IZ

https://izlik.org/JA63HG74LW

Cite

RIS / Bibtex

AMA

1.Çiçek Z, Akıllıoğlu K, Yaşar ZG, Doğan A. Intracellular Angiotensin-II Measurement in Streptozotocin-Induced Rat Vascular Smooth Muscle Cells and Its Relationship with Angiotensin-II Receptors. Eur Res J. 2025;(Advanced Online Publication):407-421. doi:10.18621/eurj.1760073