Department of Life Sciences and Technology, Xinxiang Medical University, Xinxiang, Henan, China.
Synthetic Biology Remaking Engineering and Application Laboratory, Xinxiang, Henan, China.
Cardiovasc Ther. 2017 Oct;35(5). doi: 10.1111/1755-5922.12282.
Pitavastatin (Pit) has been proved to efficiently inhibit the onset and progression of atherosclerosis. However, the mechanism by which Pit exerts nonlipid-related effects, such as antiinflammatory actions, is not quite clear. Our study aimed at investigating the effect of Pit on the expression of endothelial NO synthase (eNOS) and miR-155 in LPS-stimulated HUVECs to reveal the antiinflammatory mechanism of pitavastatin.
HUVECs were isolated from newborn umbilical cords and used in the experiments at passages 2-5. Cells were treated with LPS (0.05, 0.1, 1 μg/L) or LPS (0.1 μg/L)+Pit (0.01, 0.1, 1 μmol/L), untreated cells were used as control. For LPS+Pit induction, cells were firstly incubated with Pit for 1 hour before coincubation with LPS for 24 hours. eNOS mRNA and miR-155 were detected by RT-PCR, and Western blotting was used to detect protein expression of eNOS.
Treatment of HUVECs with LPS enhanced the expression of miR-155 and reduced the expression of eNOS in mRNA and protein level in a dose-dependent manner as revealed by RT-PCR and Western blotting, respectively. Pitavastatin ameliorated LPS-induced endothelial dysfunction through upregulation of eNOS expression and downregulation of miR-155 expression.
Pitavastatin increases eNOS expression and inhibits of LPS-induced miR-155 expression.
匹伐他汀（Pit）已被证明能有效抑制动脉粥样硬化的发生和进展。然而，匹伐他汀发挥非脂质相关作用（如抗炎作用）的机制尚不清楚。本研究旨在探讨匹伐他汀对 LPS 刺激的 HUVECs 中内皮型一氧化氮合酶（eNOS）和 miR-155 表达的影响，以揭示匹伐他汀的抗炎机制。
从新生儿脐带中分离出 HUVECs，并在第 2-5 代用于实验。用 LPS（0.05、0.1、1μg/L）或 LPS（0.1μg/L）+匹伐他汀（0.01、0.1、1μmol/L）处理细胞，未处理的细胞作为对照。对于 LPS+匹伐他汀诱导，细胞先用匹伐他汀孵育 1 小时，然后与 LPS 共孵育 24 小时。通过 RT-PCR 检测 eNOS mRNA，Western blot 检测 eNOS 蛋白表达，检测 miR-155 的表达。
LPS 处理 HUVECs 可剂量依赖性地上调 miR-155 的表达，下调 eNOS 在 mRNA 和蛋白水平的表达，分别通过 RT-PCR 和 Western blot 证实。匹伐他汀通过上调 eNOS 表达和下调 LPS 诱导的 miR-155 表达改善内皮功能障碍。
匹伐他汀增加 eNOS 表达并抑制 LPS 诱导的 miR-155 表达。
