Sinha Institute of Medical Science and Technology, Garia, Kolkata-700084, India.
Department of Botany, University of Calcutta, 35 Ballygunge Circular Road, Kolkata-700019, India.
Curr Mol Med. 2018;18(10):672-678. doi: 10.2174/1566524019666190208095754.
An increase in the level of cytokines like TNF-α and IL-6 causes the inflammatory surge in acute ischemic heart disease (AIHD).
A high-level dermcidin isoform-2 (DCN-2) occurrence in AIHD was subjected to determine a possible regulation of cytokines expression. The effect of estrogen to counteract the inflammatory response was determined.
Blood was collected from AIHD patients and normal volunteers with consent. Nitric oxide (NO) synthesis was done with methemoglobin method.TNF-α and IL-6 expression were determined by ELISA and Western blot.
(DCN-2) incubation with 120nM to the normal neutrophil solution for 2h resulted in the increase of TNF-α from 3.82±1.53pg/ml to 20.7±6.9pg/ml and IL-6 from 3.27±1.52pg/ml to 47.07±3.4pg/ml. In AIHD patients, the cytokine level was18.3- 27.3pg/ml, with a median value 21.86pg/ml (TNF-α) and IL-6 level was 23.54- 52.73pg/ml, with a median value 42.16pg/ml. Treatment with 0.6nM estriol, a kind of female steroid hormone estrogen for 45min decreased the elevated cytokine level in 120nM DCN-2 treated normal neutrophils. DCN-2 induced TNF-α synthesis in neutrophils was further determined by Western blot technique with a thickened band intensity of TNF-α. Estriol (0.6nM) treatment also influenced the DCN-2 induced inhibition of nitric oxide (NO) synthesis from 0nmol NO/ml to 0.56nmol/ml. The subsequent reduction of TNF-α level correlates the increase of NO level.
In conclusion, the stress-induced DCN-2 production in AIHD propagates the inflammatory response. Steroid molecule like estriol plays a protective role by reducing DCN-2 responses in the NO synthesis.
TNF-α和 IL-6 等细胞因子水平的升高导致急性缺血性心脏病 (AIHD) 中的炎症激增。
检测 AIHD 中高水平的 Dermcidin 同工型-2 (DCN-2) 的出现，以确定其对细胞因子表达的可能调节作用。并确定雌激素对炎症反应的拮抗作用。
征得 AIHD 患者和正常志愿者的同意后采集血液。用高铁血红蛋白法测定一氧化氮 (NO) 合成。通过 ELISA 和 Western blot 测定 TNF-α和 IL-6 的表达。
(DCN-2) 孵育 120nM 正常中性粒细胞溶液 2 小时，导致 TNF-α从 3.82±1.53pg/ml 增加至 20.7±6.9pg/ml，IL-6 从 3.27±1.52pg/ml 增加至 47.07±3.4pg/ml。在 AIHD 患者中，细胞因子水平为 18.3-27.3pg/ml，中位数为 21.86pg/ml（TNF-α），IL-6 水平为 23.54-52.73pg/ml，中位数为 42.16pg/ml。用 0.6nM 雌三醇（一种女性甾体激素雌激素）处理 45 分钟，可降低 120nM DCN-2 处理的正常中性粒细胞中升高的细胞因子水平。通过 Western blot 技术进一步确定 DCN-2 诱导中性粒细胞中 TNF-α的合成，TNF-α条带强度变厚。雌三醇（0.6nM）处理还影响 DCN-2 诱导的一氧化氮（NO）合成从 0nmol NO/ml 减少到 0.56nmol/ml。随后 TNF-α水平的降低与 NO 水平的增加相关。
总之，AIHD 中应激诱导的 DCN-2 产生会加剧炎症反应。甾体分子如雌三醇通过减少 DCN-2 对 NO 合成的反应，发挥保护作用。
