The role of miRNA in the formation of oxidative stress-induced endothelial dysfunction
Turkey (TurkMSIC) - Mustafa Kemal University, Antakya-Hatay
Department of Medical Biology, Mustafa Kemal University, Faculty of Medicine, Serinyol Mh., 31100 Hatay, Turkey
Assoc. prof. Meral Urhan Kucuk
Assoc. prof. Meral Urhan Kucuk
Turkish, English
4 weeks
Cities/Months Jan Feb Mar Apr May Jun Jul Augt Sep Oct Nov Dec
Yes Yes Yes Yes Yes No No No No No No Yes
Type of Research Project
- Basic science
What is the background of the project?
Please, provide the background of the project (Provide information about the project; min. 10 lines, max. 25 lines) Endothelium is the tissue that enlies the innermost part of hollow structures in the body, first and foremost the cardiovascular system. These cells are the primary regulators of vascular permeability, coagulability and inflammatory reactions around their habitat, and adhesion of various cells that need to interact with the tissues beyond the circulation. Imbalances in these functions are collectively called ‘’Endothelial dysfunction’’ (ED) and are the major pathological mechanism behind many diseases including atherosclerosis, hypertension and diabetes. Thus, it is imperative the mechanisms behind endothelial dysfunction be elucidated. ROS (reactive oxygen species) are reactive (aka. ‘’bond forming’’) intermediates of molecular oxygen metabolism with important roles within the cell. However, the imbalance of ROS production and clearance via antioxidant mechanisms is of primary importance in the formation of ED. Recent literature has suggested an effect of miRNA (micro Ribonucleic Acid) on the formation of ED by inhibiting the function of mRNA(messenger RNA), thus reducing the rate of cellular metabolism. The aim of this study is to investigate the role of miRNAs on the inhibition of mRNA expression of genes encoding proteins, and the effects of this phenomenon on laboratory induced ED and associated comorbities.
What is the aim of the project?
The aim of this study was to investigate the role of endothelin (ET) proteins in HUVEC (Human Umblical Vein Endothelial Cells) cells in which endothelial dysfunction is induced by exposure to hydrogen peroxide (H2O2) without any other cellular stimuli in vitro. -1), eNOS, vascular adhesion molecule 1 (VCAM 1) and plasminogen activator inhibitor 1 (PAI-1), such as expression levels of miRNAs (miRNA-126, miRNA-146, miRNA-21, miRNA-22, miRNA -29, miRNA-615-5P).
What techniques and methods are used?
What techniques and methods are used? Mention the steps/stages in this project. (Provide examples and avoid abbreviations; which tools will be used in general in this project?) 1-Cell Culture: The experiments will be carried out in the Cell Culture Laboratories of the Medical Biology Department of Mustafa Kemal University Faculty of Medicine. The commercially available HUVEC (Human Umbilical Vein Endothelial Cell) line will be used in the experiments. Cells will be grown using appropriate medium, different doses of hydrogen peroxide (H2O2) will be applied to stimulate oxidative stress, and cell viability will be evaluated to determine the effects of oxidative stress on cell proliferation and H2O2 concentrations and times to be used in the study. 2-Gene Expression Analysis: Endothelin 1 (ET-1), endothelial nitric oxide synthase (eNOS), vascular adhesion molecule 1 (VCAM 1) and plasminogen activator inhibitor 1 (PAI-1), which play a key role in endothelial function from HUVEC cells treated with hydrogen peroxide (H2O2). Gene expression levels will be determined in Real-time quantitative polymerase chain reaction (RT-qPCR). 3-miRNA Expression Assays: It is thought that it regulates genes that play a key role in endothelial function, miRNA-126, miRNA-146a, miRNA-21 and miRNA-22 expression levels will be determined in RT-qPCR. 4-Evaluation of Data: The data obtained from the studies will be evaluated and reported with appropriate statistical methods with the help of SPSS 26 (Statistical Package for Social Sciences).
What is the role of the student?
- The student will mainly observe
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- If the project includes “lab work”
- the student will take active part in the practical aspect of the project
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
What are the tasks expected to be accomplished by the student? (Minimum 10 lines, provide detailed explanation) The following tasks will be expected to be accomplished by the student: -Being able to use an inverted microscope in order to examine the adherent cells -Being able to do cell passaging, freezing and thawing according to the cell culture studies. -Being able to use Mc Excel and SPSS 26 (Statistical Package for Social Sciences) at a fundamental level for the statistical evaluation process of the project. -Evaluating the results of the gene and miRNA expression studies in RT-qPCR with the help of the tutor and DataAssist Software.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The following topics will be provided in the form of seminars by Assoc. prof. Meral Urhan Kucuk. Lessons will be provided according to the availability of the teacher, at least once per week: -General information about adheren molecules and integrins -Cell isolation process and steps -The cell cycle, transcription and translation process -Cell culture protocols
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a scientific report
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
No spesific skills are required.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts:
- Medical students
- • Role of MicroRNA in Endothelial Dysfunction and Hypertension 2018; Miruna Nemecz Nicoleta Alexandru;