Catalpol Inhibits Homocysteine-induced Oxidation and Inflammation via Inhibiting Nox4/NF-κB and GRP78/PERK Pathways in Human Aorta Endothelial Cells.
Catalpol Inhibits Homocysteine-induced Oxidation and Inflammation via Inhibiting Nox4/NF-κB and GRP78/PERK Pathways in Human Aorta Endothelial Cells.
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Catalpol通过抑制人主动脉内皮细胞中的NOX4/NF-κB和GRP78/PERK途径抑制同型半胱氨酸诱导的氧化和炎症。
DOI：
10.1007/s10753-018-0873-9
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发表时间：
2019-03
期刊：
影响因子：
5.1
通讯作者：
Sun H
中科院分区：
文献类型：
--
作者：
Hu H;Wang C;Jin Y;Meng Q;Liu Q;Liu Z;Liu K;Liu X;Sun H
高同型半胱氨酸血症（HHCY）是动脉粥样硬化的独立危险因素，在动脉粥样硬化的发生发展中起着重要作用。梓醇是从紫菀根中提取的环烯醚萜苷，具有抗炎、抗氧化、抗肿瘤和保护多巴胺能神经元的作用。本研究旨在探讨梓醇对同型半胱氨酸（HCY）诱导的人主动脉内皮细胞（HAECs）损伤的保护作用及其机制：1。将HAEC与不同浓度的HCY（3 mM）和梓醇（7.5 μ M、15 μ M、30 μ M）一起培养24小时。(1)比色法测定MDA、GSH含量及LDH释放量。(2)流式细胞仪检测细胞内活性氧（ROS）的含量。(3)Western blotting检测Nox 4、p22 phox、ICAM-1、MCP-1、VCAM-1、IκB、细胞核p65、p65磷酸化、caspase-3、-9、bax、bcl-2和ER应激相关蛋白的表达。(4)qRT-PCR检测CHOP、ATF 4的表达。(5)采用罗丹明123染色法测定HCY处理后HAECs线粒体膜电位，激光共聚焦显微镜观察标本。2.采用DPI、PDTC和TUDCA检测Nox 4/ROS、NF-κB和内质网应激之间的相互作用。3.采用TUDCA或Nox 4 siRNA研究梓醇抑制ROS过度产生的作用是否与抑制ER应激和Nox 4表达有关。梓醇显著抑制LDH释放、MDA水平和GSH还原。梓醇可抑制HCY诱导的ROS过度产生，抑制NF-κB的转录激活以及Nox 4、ICAM-1、VCAM-1和MCP-1蛋白的过度表达。梓醇在HCY处理的HAECs中升高bcl-2蛋白表达，降低bax，caspase-3，-9蛋白表达。同时，梓醇还能抑制HCY诱导的内质网应激相关感受器GRP 78、IRE 1 α、ATF 6、P-PERK、P-eIF 2 α、CHOP和ATF 4的激活。此外，DPI或TUDCA抑制Nox 4或ER胁迫后，梓醇抑制ROS过度生成和NF-κB信号通路的程度降低。NF-κB抑制剂PDTC也可降低梓醇对Nox 4和GRP 78表达的抑制作用。Nox 4 siRNA可降低梓醇抑制ROS过度生成的作用。梓醇可能通过抑制Nox 4/NF-κB和ER应激，减轻HCY诱导的HAECs氧化损伤、细胞凋亡和炎症反应。
Hyperhomocysteinemia (HHCY) has been recognized as an independent risk factor for atherosclerosis and plays a vital role in the development of atherosclerosis. Catalpol, an iridoid glucoside extracted from the root of Rehmannia glutinosa, can produce anti-inflammatory, anti-oxidant, anti-tumor, and dopaminergic neurons protecting effects. This study aimed to determine the protecting effects of catalpol against homocysteine (HCY)-induced injuries in human aortic endothelial cells (HAECs) and uncover the underlying mechanisms: 1. HAECs were cultured with different concentrations of HCY (3 mM) and catalpol (7.5 μΜ, 15 μΜ, 30 μΜ) for 24 h. (1) The level of MDA and GSH as well as LDH release was measured with colorimetric assay. (2) Reactive oxygen species (ROS) were detected by flow cytometry analysis. (3) Western blotting analysis was performed to detect the expression of Nox4, p22phox, ICAM-1, MCP-1, VCAM-1, IκB, nucleus p65, p65 phosphorylation, caspase-3, −9, bax, bcl-2, and ER stress-related proteins. (4) The expressions of CHOP, ATF4 were measured by qRT-PCR. (5) Mitochondrial membrane potential in HCY-treated HAECs was measured by rhodamine 123 staining, and the samples were observed by confocal laser scanning microscopy. 2. DPI, PDTC, and TUDCA were used to determine the interaction among Nox4/ROS, NF-κB, and endoplasmic reticulum stress. 3. TUDCA or Nox4 siRNA were used to investigate whether the effect of catalpol inhibiting the over-production of ROS were associated with inhibiting ER stress and Nox4 expression. Catalpol significantly suppressed LDH release, MDA level, and the reduction of GSH. Catalpol reduced HCY-stimulated ROS over-generation, inhibited the NF-κB transcriptional activation as well as the protein over-expressions of Nox4, ICAM-1, VCAM-1, and MCP-1. Catalpol elevated bcl-2 protein expression and reduced bax, caspase-3, −9 protein expressions in the HCY-treated HAECs. Simultaneously, catalpol could also inhibit the activation of ER stress-associated sensors GRP78, IRE1α, ATF6, P-PERK, P-eIF2α, CHOP, and ATF4 induced by HCY. In addition, the extent of catalpol inhibiting ROS over-generation and NF-κB signaling pathway was reduced after inhibiting Nox4 or ER stress with DPI or TUDCA. The inhibitor of NF-κB PDTC also reduced the effects of catalpol inhibiting the expressions of Nox4 and GRP78. Furthermore, the effect of catalpol inhibiting the over-generation of ROS was reduced by Nox4 siRNA. Catalpol could ameliorate HCY-induced oxidation, cells apoptosis and inflammation in HAECs possibly by inhibiting Nox4/NF-κB and ER stress.
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影响因子：
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发表时间：
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影响因子：
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