ABSTRACT
Background and purpose: The 6th edition of International Diabetes Federation, 2014 shows an estimate of 387 million people with Type 2 diabetes mellitus [T2DM] worldwide, expected to rise to 592 million by 2035. T2DM is a metabolic disorder, one of the reasons being oxidative stress due to impairment in antioxidant enzymes. It leads to several complications such as micro and macrovascular diseases. Cyclooxygenase1 [COX1] enzyme is the rate limiting factor for the arachidonic pathway leading to vascular wall contraction with angiotensin II occurring in heart diseases resulting from T2DM. COX1 determines 6-Keto Prostaglandin F1alpha [6-k-PGF1alpha] level, plays a major role in vasodilation and restricts macrophage platelet aggregation. The aim of the present study was to compare the COX1 expression and level of reactive oxygen species [ROS] in T2DM patients and controls at different time periods in human macrophages in order to find a biomarker or drug target
Subjects and methods: The study subjects consisted of 100 individuals, 50 each from T2DM patients and healthy sex/age matched controls. Cell proliferation by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide [MTT] assay and ROS measurement by 2',7'-dichlorofluorescein diacetate [DCFDA] staining were performed at different time periods [24, 48, 72 h]. COX1 mRNA expression was checked by relative quantification method after real-time polymerase chain reaction [RT-PCR]
Results: The MTT assay showed that cell viability was significantly higher at 48 h [P < 0.05]. ROS production was found to be lowest at 24 h by DCFDA staining. ROS levels were raised in T2DM patients as compared to controls. The quantitative RT-PCR analysis showed that the COX1 expression was higher in T2DM patients as compared to healthy controls although not significant [P > 0.05]
Conclusion: Although COX1 is known to be a "housekeeping" gene, our study showed that its expression can be correlated with the disease condition and be used as a marker. However, further studies are required in more number of samples from other ethnic populations to confirm the findings