Effect of Vitamin D3 in combination with Omega‐3 Polyunsaturated Fatty Acids on NETosis in Type 2 Diabetes Mellitus Patients

Abstract

An understanding of the consequences of oxidative/halogenative stress triggered by neutrophil activation is impossible without considering NETosis. NETosis, formation of neutrophil extracellular traps (NETs), is known to promote microthrombus formation and impair wound healing in type 2 diabetes mellitus (T2DM) patients. Therefore, there is a need to search for drugs and treatment approaches that could prevent excessive NET formation. We aimed to evaluate the effect of vitamin D3 in combination with omega-3 polyunsaturated fatty acids (vitamin D3/omega-3 PUFAs) on NETosis in T2DM patients with purulent necrotizing lesions of the lower extremities. Patients and healthy subjects had vitamin D3 deficiency. Patients received, beyond standard treatment, 6000 IU of vitamin D3 and 480 mg of omega-3 PUFAs, and healthy subjects 1000 IU of vitamin D3 and 240 mg of omega-3 PUFAs daily for seven days. Neutrophil activation in ex vivo blood by phorbol-12-myristate-13-acetate (PMA) was used as a NETosis model. The percentage of blood NETs relative to leukocytes (NETbackground) before vitamin D3/omega-3 PUFA supplementation was 3.2%-4.9% in healthy subjects and 1.7%-10.8% in patients. These values rose, respectively, to 7.7%-9.1% and 4.0%-17.9% upon PMA-induced NETosis. In addition, the leukocyte count decreased by 700-1300 per 1 μL in healthy subjects and 700-4000 per 1 μL in patients. For both patients and healthy subjects, taking vitamin D3/omega-3 PUFAs had no effect on NETbackground but completely inhibited PMA-induced NET formation, though neutrophils exhibited morphological features of activation. Also, leukocyte loss was reduced (to 500 per 1 μL). For patients on standard treatment alone, changes occurred neither in background NETs and leukocytes nor in their amount after PMA stimulation. The decreased ability of neutrophils to generate NETs, which can be achieved by vitamin D3/omega-3 PUFA supplementation, could have a positive effect on wound healing in T2DM patients and reduce the incidence and severity of complications.

Calcifediol boosts the efficacy of ChAdOx1 nCoV-19 (COVISHIELD) vaccine via upregulation of key genes associated with memory T cell responses

“…Vitamin D has also been reported in other diseases to reduce the formation of NETs (19,20) including the treatment of SLE (21).…”

Calcifediol boosts efficacy of ChAdOx1 nCoV-19 vaccine by upregulating genes promoting memory T cell responses

“…Neutrophils are essential for innate immunity. However, COVID-19 disease, results in high levels of circulating NETs causing a reduction in blood flow in the capillaries leading to build-up of NETs in the lungs and other organs causing multiple organ failures 26 . Therefore, calcifediol appeared to modulate NETosis which might be important in curbing exaggerated inflammatory responses during the initial stages of secondary COVID infection.…”

Methylglyoxal-Modified Human Serum Albumin Binds to Leukocyte Myeloperoxidase and Inhibits its Enzymatic Activity

“…The smears were examined using a Motic B3 microscope (Motic Asia, Hong Kong). NET-like structures per 300–500 leukocytes were counted in the middle third of the smear, and the NET-to-leukocyte percentage ratio was calculated [ 15 ]. DNA in NET-like structures was visualized with Hoechst 33,342 staining (see Figure S2 in Supplementary Materials ) using the Nikon Eclipse Ni-E fluorescent microscope (Nikon, Tokyo, Japan).…”