Role of LOX-1 and ROS in oxidized low-density lipoprotein induced epithelial-mesenchymal transition of NRK52E.
Role of LOX-1 and ROS in oxidized low-density lipoprotein induced epithelial-mesenchymal transition of NRK52E.
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DOI：
10.1186/1476-511x-9-120
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发表时间：
2010-10-19
影响因子：
4.5
通讯作者：
Yang H
中科院分区：
文献类型：
--
作者：
Wang R;Ding G;Liang W;Chen C;Yang H
探讨氧化型低密度脂蛋白受体-1(LOX-1)在氧化型低密度脂蛋白(OX-LDL)诱导的肾小管上皮-间充质转化(TEMT)中的作用及其机制。用氧化低密度脂蛋白(0、25、50和100μg/ml)孵育NRK-52E细胞24小时，或用LOX-1受体多聚肌苷酸(Poly I)和角叉菜胶预处理或用抗氧化剂N-乙酰-L半胱氨酸(NAC)处理后，用实时定量聚合酶链式反应、Western blotting分析、免疫荧光和激光共聚焦扫描显微镜检测LOX-I、E-钙粘素、α-平滑肌肌动蛋白(α-SMA)和活性氧(ROS)的表达。氧化低密度脂蛋白在0~100μg/ml范围内呈剂量依赖性增加LOX-1mRNA和蛋白的表达(P<0.05)。随着LOX-1蛋白水平的升高，脂质摄入量、ROS生成和α-SMA表达增加，而E-钙粘素水平下降。多糖I、角叉菜胶或NaC可显著抑制氧化低密度脂蛋白诱导的LOX-1表达、α-SMA表达、脂质摄取和ROS生成，并逆转氧化低密度脂蛋白诱导的E-钙粘蛋白表达下降。同时，ROS的产生与LOX-1表达的增加有关。α-SMA表达与ROS生成和LOX-1表达呈正相关，E-钙粘蛋白表达与ROS生成和LOX-1表达呈负相关。LOX-1和ROS可能在氧化低密度脂蛋白诱导的NRK52E上皮间充质转化中起重要作用。
To investigate the effect of oxidized low density lipoprotein receptor-1 (LOX-1) on tubular epithelial-mesenchymal transition (TEMT) induced by oxidized low-density lipoprotein (ox-LDL) and its mechanism. NRK-52E cells were incubated with ox-LDL (0, 25, 50, and 100 μg/ml) for 24 hours or pre-treated with the chemical inhibitor of the LOX-1 receptor polyinosinic acid (poly I) and carrageenan or the antioxidant N-acetyl-L-cysteine (NAC), the cells were then exposed to 50 μg/ml of ox-LDL.The expression of LOX-I, E-cadherin, α-smooth muscle actin (α-SMA) and reactive oxygen species (ROS) were analyzed by real-time PCR, western blotting analysis, immunofluorescence and confocal laser scanning microscopy. Ox-LDL increased the expression of LOX-1 mRNA and protein in a dose-dependent manner from 0 to 100 μg/ml (P < 0.05). Following the increase in the LOX-1 protein level, the lipid intake, ROS generation and α-SMA expression increased; however, the E-cadherin level decreased. The pre-treatment with poly I or carrageenan or NAC significantly inhibited the LOX-1 expression, α-SMA expression, the lipid intake and ROS generation and reversed decrease of E-cadherin expression induced by ox-LDL. Meanwhile, the ROS generation were associated with a increase in the LOX-1 expression. The α-SMA expression was positively correlated with the ROS generation and LOX-1 expression, and the E-cadherin expression was negatively correlated with the ROS generation and LOX-1 expression. LOX-1 and ROS may play a important role in epithelial-mesenchymal transition of NRK52E induced by OX-LDL.