Ridzuan Noridzzaida, John Cini Mathew, Sandrasaigaran Pratheep, Maqbool Maryam, Liew Lee Chuen, Lim Jonathan, Ramasamy Rajesh
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.
World J Diabetes. 2016 Jul 10;7(13):271-8. doi: 10.4239/wjd.v7.i13.271.
To assess the amount and pattern of reactive oxygen species (ROS) production in diabetic patient-derived neutrophils.
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.
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.
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.
评估糖尿病患者来源的中性粒细胞中活性氧（ROS）的产生量及模式。
采集2型糖尿病（DM）患者和志愿者（对照组）的血样，进行中性粒细胞分离，并采用化学发光法评估中性粒细胞的氧化爆发。使用佛波酯（PMA）激活中性粒细胞，未激活的中性粒细胞作为阴性对照。将细胞悬液转移至含有PMA和鲁米诺的1.5 mL艾本德管中，获取化学发光读数。反应混合物轻轻涡旋后置于发光计中5分钟。
我们的结果显示，在静息状态下，正常非糖尿病个体中ROS的分泌相较于糖尿病患者相对较低。然而，时间尺度观察表明，非糖尿病个体中分泌的ROS随时间相应下降，但在糖尿病患者中未检测到这种下降，在所有时间点，ROS的分泌均维持在相似水平。这项初步研究表明，在静息和激活状态下，DM患者中ROS的产生均显著高于非糖尿病受试者。
DM可能影响中性粒细胞的呼吸爆发活性，ROS产生增加可能是糖尿病相关并发症的一个加重因素。
