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©The Author(s) 2016. Published by Baishideng Publishing Group Inc. All rights reserved.
Preliminary study on overproduction of reactive oxygen species by neutrophils in diabetes mellitus
Noridzzaida Ridzuan, Cini Mathew John, Pratheep Sandrasaigaran, Maryam Maqbool, Lee Chuen Liew, Jonathan Lim, Rajesh Ramasamy
Noridzzaida Ridzuan, Cini Mathew John, Pratheep Sandrasaigaran, Maryam Maqbool, Lee Chuen Liew, Rajesh Ramasamy, Immunology Laboratory, Department of Pathology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia
Noridzzaida Ridzuan, Maryam Maqbool, Rajesh Ramasamy, Stem Cell Research Laboratory, Genetic and Regenerative Medicine Research Center, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia
Cini Mathew John, Department of Physiology and Pharmacology, Faculty of Medicine, University of Calgary, Calgary, AB T2N 4N1, Canada
Jonathan Lim, Department of Medicine, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia
Author contributions: All the authors contributed to this manuscript.
Institutional review board statement: No institutional review board statement because this is a preliminary study conducted with a very limited number of patients based on their voluntary contribution and thus ethical approval from institute was not sought. However, the handling of samples was carried out in accordance with ethical values and standard procedures.
Informed consent statement: Informed consent was obtained from the voluntarily recruited patients. They were briefed on the purpose of the study and its implication prior to donating 10 mL of peripheral blood. History of medication and management of diabetes was collected with permission of the relevant patients.
Conflict-of-interest statement: No conflict of interest declared by any of the authors.
Data sharing statement: No data sharing as this manuscript and the data were not published elsewhere.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Rajesh Ramasamy, PhD, Immunology Laboratory, Department of Pathology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang Hospital Street, Serdang 43400, Selangor, Malaysia.
rajesh@upm.edu.my
Telephone: +603-89-472377 Fax: +603-89-412787
Received: February 29, 2016
Peer-review started: March 1, 2016
First decision: March 22, 2016
Revised: May 26, 2016
Accepted: June 1, 2016
Article in press: June 3, 2016
Published online: July 10, 2016
Processing time: 127 Days and 18.6 Hours
AIM: To assess the amount and pattern of reactive oxygen species (ROS) production in diabetic patient-derived neutrophils.
METHODS: Blood samples from type 2 diabetes mellitus (DM) patients and volunteers (controls) were subjected to neutrophil isolation and the assessment of neutrophil oxidative burst using chemiluminescence assay. Neutrophils were activated by using phorbol myristate acetate (PMA) and neutrophils without activation were kept as a negative control. The chemiluminescence readings were obtained by transferring cell suspension into a 1.5 mL Eppendorf tube, with PMA and luminol. Reaction mixtures were gently vortexed and placed inside luminometer for a duration of 5 min.
RESULTS: Our results showed that in the resting condition, the secretion of ROS in normal non-diabetic individuals was relatively low compared to diabetic patients. However, the time scale observation revealed that the secreted ROS declined accordingly with time in non-diabetic individuals, yet such a reduction was not detected in diabetic patients where at all the time points, the secretion of ROS was maintained at similar magnitudes. This preliminary study demonstrated that ROS production was significantly higher in patients with DM compared to non-diabetic subjects in both resting and activated conditions.
CONCLUSION: The respiratory burst activity of neutrophils could be affected by DM and the elevation of ROS production might be an aggravating factor in diabetic-related complications.
Core tip: This is a preliminary study that investigates the activation status of peripheral blood-derived neutrophils in type 2 diabetes. This study clearly indicated that the neutrophils from type 2 diabetic patients are highly activated upon in vitro stimulation and hence produce greater amounts of reactive oxygen species (ROS) compared to a normal individual. Release of a greater volume of ROS could serve as an additional risk for end organ injury in type 2 diabetes mellitus.
