Abstract

Context

Recent studies demonstrated the anti-atherosclerotic efficacy of cyclodextrin. However, it remains unclear whether cyclodextrin exerts the anti-atherosclerotic effect via regulating monocyte-endothelial adhesion.

Objective

To answer that question by recruiting methyl-β-cyclodextrin (MβCD) as a cyclodextrin representative.

Materials and methods

Human umbilical vein endothelial cells (HUVECs) were not treated, or treated with 1 µg/mL liposaccharide (LPS) or 50 µg/mL oxidized low-density lipoprotein (oxLDL) for 12 h, 5 mM MβCD for 1 h, and LPS/oxLDL (1 and 50 µg/mL, respectively for 12 h) plus MβCD (5 mM for 1 h), respectively. The effects of MβCD on LPS/oxLDL-triggered monocyte-endothelial adhesion and related molecules in signalling pathways were evaluated via confocal microscopy, flow cytometry, RT-PCR, western blotting, and cell adhesion assay.

Results

MβCD with an IC₅₀ of 27.66 mM (1 h treatment) exerted no significant cytotoxicity at ≤5 mM for ≤2 h. Compared with the control, both LPS and oxLDL induced an ∼2–3-fold increase in adhesion molecule expression (ICAM-1 and VCAM-1 at protein and mRNA levels) and NF-κB phosphorylation (p-NF-κB/pP65), an increase in IκB kinase (IKK), and a decrease in phosphorylated protein kinase B (p-Akt), respectively. Moreover, more monocytes (2-fold higher for LPS and 15% higher for oxLDL) were attached on LPS/oxLDL-stimulated HUVECs. 5 mM MβCD reversed the LPS/oxLDL-induced changes back to the control levels.

Conclusions

MβCD significantly suppresses the LPS/oxLDL-triggered monocyte-endothelial adhesion by downregulating adhesion molecule expression probably via LPS-IKK-NF-κB or oxLDL-Akt-NF-κB pathway. This study demonstrates a potential mechanism of the anti-atherosclerotic efficacy of cyclodextrin from the angle of monocyte-endothelial adhesion.

中文翻译：

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甲基-β-环糊精抑制由脂多糖 (LPS) 或氧化低密度脂蛋白 (oxLDL) 引发的单核细胞-内皮细胞粘附

摘要

语境

最近的研究证明了环糊精的抗动脉粥样硬化功效。然而，目前尚不清楚环糊精是否通过调节单核细胞-内皮细胞粘附发挥抗动脉粥样硬化作用。

客观的

通过招募甲基-β-环糊精 (MβCD) 作为环糊精代表来回答这个问题。

材料和方法

人脐静脉内皮细胞 (HUVEC) 未处理，或用 1 µg/mL 脂糖 (LPS) 或 50 µg/mL 氧化低密度脂蛋白 (oxLDL) 处理 12 小时、5 mM MβCD 处理 1 小时和 LPS/ oxLDL（1 和 50 µg/mL，分别为 12 小时）加 MβCD（5 mM，1 小时），分别。通过共聚焦显微镜、流式细胞术、RT-​​PCR、蛋白质印迹和细胞粘附试验评估了 MβCD 对 LPS/oxLDL 触发的单核细胞-内皮细胞粘附和信号通路中相关分子的影响。

结果

具有 27.66 mM（1 小时处理）的 IC ₅₀的 MβCD 在≤5 mM ≤2 小时内没有表现出显着的细胞毒性。与对照相比，LPS 和 oxLDL 均诱导粘附分子表达（蛋白质和 mRNA 水平的 ICAM-1 和 VCAM-1）和 NF-κB 磷酸化（p-NF-κB/pP65）增加约 2-3 倍，分别增加 IκB 激酶 (IKK) 和磷酸化蛋白激酶 B (p-Akt) 的减少。此外，更多的单核细胞（LPS 高 2 倍，oxLDL 高 15%）附着在 LPS/oxLDL 刺激的 HUVEC 上。5 mM MβCD 将 LPS/oxLDL 诱导的变化逆转回控制水平。

结论

MβCD 可能通过 LPS-IKK-NF-κB 或 oxLDL-Akt-NF-κB 通路下调粘附分子表达，从而显着抑制 LPS/oxLDL 触发的单核细胞-内皮细胞粘附。该研究从单核细胞-内皮细胞粘附的角度证明了环糊精抗动脉粥样硬化功效的潜在机制。