Overexpression of steroidogenic acute regulatory protein in rat aortic endothelial cells attenuates palmitic acid-induced inflammation and reduction in nitric oxide bioavailability.
Overexpression of steroidogenic acute regulatory protein in rat aortic endothelial cells attenuates palmitic acid-induced inflammation and reduction in nitric oxide bioavailability.
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大鼠主动脉内皮细胞中类固醇生成急性调节蛋白的过度表达可减轻棕榈酸诱导的炎症和一氧化氮生物利用度的降低
DOI：
10.1186/1475-2840-11-144
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发表时间：
2012-11-21
影响因子：
9.3
通讯作者：
Ning Y
中科院分区：
文献类型：
--
作者：
Tian D;Qiu Y;Zhan Y;Li X;Zhi X;Wang X;Yin L;Ning Y
背景内皮功能障碍是动脉粥样硬化和其他心血管疾病发病的重要证据。血脂紊乱是这些疾病中内皮功能障碍的主要危险因素之一。类固醇生成急性调节蛋白（StAR）是胆固醇转运蛋白之一，在维持细胞内脂质平衡中起重要作用。然而，StAR对内皮功能障碍的影响还不清楚。棕榈酸（PA）可降低内皮型一氧化氮合酶（eNOS）活性，诱导炎症反应，这两种反应都是内皮功能障碍的原因，在内皮细胞培养模型中，通过腺病毒感染将StAR基因导入原代大鼠主动脉内皮细胞。通过实时荧光定量PCR和Western blotting检测相关基因和蛋白的表达水平，探讨其作用机制。游离脂肪酸和胆固醇定量试剂盒用于检测细胞总游离脂肪酸和胆固醇。用ELISA法和经典的Griess试剂法分别测定大鼠主动脉内皮细胞中炎症因子和一氧化氮的水平。StAR过表达后，细胞培养上清中IL-1 β、TNF α、IL-6和VCAM-1的mRNA水平及IL-1 β、TNF α和IL-6的蛋白水平均显著降低，这是由于StAR抑制了NF κ B核转位和活化。此外，StAR过表达通过保护pAkt/peNOS/NO通路的生物活性来减弱PA诱导的一氧化氮生物利用度的降低。此外，参与脂质代谢的关键基因在StAR过表达后大大减少。为了研究潜在的机制，浅蓝菌素和洛伐他汀，脂肪酸和胆固醇合成酶的抑制剂，在PA处理之前加入。结果表明浅蓝菌素和洛伐他汀都具有与StAR过表达相似的作用。另一方面，StAR的作用被抑制时，siRNA被引入，以减少StAR expressions.ConclusionsOur结果表明，StAR衰减脂质合成和摄取以及PA诱导的炎症和减少NO生物利用度在主动脉内皮细胞。StAR可通过降低细胞内脂质水平改善PA诱导的内皮功能障碍。
BackgroundEndothelial dysfunction is a well documented evidence for the onset of atherosclerosis and other cardiovascular diseases. Lipids disorder is among the main risk factors for endothelial dysfunction in these diseases. Steroidogenic acute regulatory protein (StAR), one of the cholesterol transporters, plays an important role in the maintenance of intracellular lipid homeostasis. However, the effect of StAR on endothelial dysfunction is not well understood. Palmitic acid (PA) has been shown to decrease eNOS activity and induce inflammation, both are the causes of endothelial dysfunction, in an endothelial cell culture model.MethodsStAR gene was introduced into primary rat aortic endothelial cells by adenovirus infection. Real-time PCR and Western blotting were performed to determine the relative genes and proteins expression level to elucidate the underlying mechanism. The free fatty acid and cholesterol quantification kits were used to detect total cellular free fatty acid and cholesterol. The levels of inflammatory factors and nitric oxide were determined by ELISA and classic Griess reagent methods respectively.ResultsWe successfully overexpressed StAR in primary rat aortic endothelial cells. Following StAR overexpression, mRNA levels of IL-1β, TNFα, IL6 and VCAM-1 and protein levels of IL-1β, , TNFα and IL-6 in culture supernatant were significantly decreased, which duing to blocke NFκB nuclear translocation and activation. Moreover, StAR overexpression attenuated the PA-induced reduction of nitric oxide bioavailability by protecting the bioactivity of pAkt/peNOS/NO pathway. Furthermore, the key genes involved in lipid metabolism were greatly reduced following StAR overexpression. In order to investigate the underlying mechanism, cerulenin and lovastatin, the inhibitor of fatty acid and cholesterol synthase, were added prior to PA treatment. The results showed that both cerulenin and lovastatin had a similar effect as StAR overexpression. On the other hand, the role of StAR was inhibited when siRNA was introduced to reduce StAR expression.ConclusionsOur results showed that StAR attenuated lipid synthesis and uptake as well as PA-induced inflammation and reduction in NO bioavailability in aortic endothelial cells. StAR can ameliorate endothelial dysfunction induced by PA via reducing the intracellular lipid levels.
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影响因子：
8.3
作者：
Azekoshi, Yoko;Yasu, Takanori;Ueda, Shinichiro
通讯作者：
Ueda, Shinichiro
影响因子：
4.2
作者：
Ajuwon, KM;Spurlock, ME
通讯作者：
Spurlock, ME
影响因子：
9.3
作者：
Aragonès G;Saavedra P;Heras M;Cabré A;Girona J;Masana L
通讯作者：
Masana L
影响因子：
6.7
作者：
Ma, Y.;Ren, S.;Yin, L.
通讯作者：
Yin, L.
DOI：
10.1152/ajpheart.00292.2006
发表时间：
2006-09-01
影响因子：
4.8
作者：
Feletou, Michel;Vanhoutte, Paul M.
通讯作者：
Vanhoutte, Paul M.