Handono Kusworini, Sidarta Yona One, Pradana Benny A, Nugroho Radhitio A, Hartono Ivan A, Kalim Handono, Endharti Agustina T
Department of Clinical Pathology, Faculty of Medicine, Universitas Brawijaya - Saiful Anwar Hospital, Malang, Indonesia.
Acta Med Indones. 2014 Jul;46(3):189-98.
to investigate the effects of Vitamin D calcitriol/1,25(OH)2D3 on NETosis in systemic lupus erythematosus (SLE) patients with hypovitamin D.
neutrophlis of five SLE patients with hypovitamin D were divided into 4 groups, P0 (0 nM/control), P1 (1 nM), P2 (10 nM), and P3 (100 nM) as cultured samples. Phorbol Myristate Acetate (PMA) was used to stimulate NETs formation. The supernatant was separated and cocultured with HUVECs. Externalization of Neutrophil Elastase (NE) and Myeloperoxidase (MPO) during NETosis was measured by immunofluorescence and ELISA respectively. Early and late apoptosis of endothelial cell was measured by flowcytometry using cell death kit (Annexin V and PI antibody).
this study showed significant decrease in early apoptosis with 10 nM of 1,25(OH)2D3 compared to control group. Significance of NE externalization found in all treatment groups (p<0.05), while MPO absorbance in the same tendency but not statistically significant. Further analysis also found a moderate positive correlation between NE externalizations with early apoptosis.
vitamin D 1,25(OH)2D3 could reduce endothelial damage by decreasing NETosis activity. This result may reveal the possibility of Vitamin D as supplementary therapy for SLE patients with hypovitamin D to prevent endothelial damage.
研究维生素D骨化三醇/1,25(OH)₂D₃对维生素D缺乏的系统性红斑狼疮（SLE）患者中性粒细胞胞外诱捕网形成（NETosis）的影响。
将5例维生素D缺乏的SLE患者的中性粒细胞分为4组，即P0（0 nM/对照组）、P1（1 nM）、P2（10 nM）和P3（100 nM）作为培养样本。用佛波酯（PMA）刺激NETs形成。分离上清液并与人类脐静脉内皮细胞（HUVECs）共培养。分别通过免疫荧光和酶联免疫吸附测定（ELISA）检测NETosis过程中中性粒细胞弹性蛋白酶（NE）和髓过氧化物酶（MPO）的外化情况。使用细胞死亡试剂盒（膜联蛋白V和碘化丙啶抗体）通过流式细胞术检测内皮细胞的早期和晚期凋亡。
本研究表明，与对照组相比，10 nM的1,25(OH)₂D₃可使早期凋亡显著减少。在所有治疗组中均发现NE外化具有统计学意义（p<0.05），而MPO吸光度呈相同趋势但无统计学意义。进一步分析还发现NE外化与早期凋亡之间存在中度正相关。
维生素D 1,25(OH)₂D₃可通过降低NETosis活性来减少内皮损伤。这一结果可能揭示了维生素D作为维生素D缺乏的SLE患者补充治疗以预防内皮损伤的可能性。
