Preliminary study on overproduction of reactive oxygen species by neutrophils in diabetes mellitus

Figure 1 Morphological analysis of isolated neutrophils. Isolated neutrophils were smeared on a glass slide and labeled with Leishman stain. Under the 20 × magnification, cells appeared homogeneous (A) and 40 × magnification exhibited the multi-lobulated nucleus (B).

Figure 2 Temporal analysis of reactive oxygen species secreted by neutrophils of non-diabetic and diabetic subjects. Isolated neutrophils were measured for ROS secretion via chemiluminescence method at every 10 s over a period of 50 s. Neutrophils isolated from diabetic patients showed a higher and consistent secretion of ROS in resting condition (A). PMA stimulation elevated the ROS secretion from neutrophils of both non-diabetic and diabetic subjects. Neutrophils from diabetic patients produced a higher amount of ROS compared to non-diabetic subjects. ROS production was determined at every 10 s beginning from the administration of PMA to the time point where the ROS production reached a maximal level and started to decline (B). Data expressed as mean ± SD and the statistical significance was determined at ^bP value ≤ 0.001. ROS: Reactive oxygen species; PMA: Phorbol myristate acetate.

Figure 3 Total reactive oxygen species produced by neutrophils of non-diabetic and diabetic subjects. Neutrophils isolated from non-diabetic individuals and diabetic patients subjected for ROS secretion at resting condition and upon stimulation with PMA. The maximal production of ROS at any time point over a period of 50 s was recorded and compared. Neutrophils from diabetic patients showed a higher production of ROS at resting and activated conditions. Data expressed as mean ± SD and the statistical significance was determined at ^aP value ≤ 0.05. ROS: Reactive oxygen species; PMA: Phorbol myristate acetate.