Inhibition of connexin 43 attenuates oxidative stress and apoptosis in human umbilical vein endothelial cells
Inhibition of connexin 43 attenuates oxidative stress and apoptosis in human umbilical vein endothelial cells
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DOI：
10.1186/s12890-019-1036-y
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发表时间：
2020-01-21
影响因子：
3.1
通讯作者：
Luo, Liang
中科院分区：
文献类型：
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作者：
Ma, Jia-wei;Ji, Dan-dan;Luo, Liang
背景以往的研究表明连接蛋白43（connexin 43，Cx43）通过影响线粒体功能在细胞凋亡调控中发挥重要作用。本研究旨在探讨脂多糖（lipopolysaccharide，LPS）诱导的人脐静脉内皮细胞（human umbilical vein endothelial cells，HUVECs）氧化应激与细胞凋亡的关系。方法采用Western blot方法检测线粒体Cx43（MtCx 43）蛋白水平及磷酸化水平。Gap 19是一种选择性Cx43抑制剂，用于检测Cx43对LPS诱导的HUVECs氧化应激和凋亡的影响。通过定量聚合酶链反应（qPCR）和Western blot检测与氧化应激相关的调控基因的表达。流式细胞术检测细胞凋亡。结果LPS刺激可导致MtCx 43和p-MtCx 43表达增加。有趣的是，Gap 19拮抗由LPS或Cx43过表达诱导的谷胱甘肽S-转移酶Zeta 1（GSTZ 1）和细胞色素B α β（CYBB）的上调，以及抗氧化剂1（ATOX 1）、谷胱甘肽合成酶（GSS）和血红素加氧酶1（HMOX 1）的下调。此外，用Gap 19处理后，由LPS或Cx43过表达引起的活性氧（ROS）和细胞凋亡的产生增加减少。结论选择性抑制Cx43半通道可保护HUVECs免受LPS诱导的凋亡，这可能是通过减少氧化应激的产生。
Background Previous studies demonstrated an important role for connexin 43 (Cx43) in the regulation of apoptosis by influencing mitochondrial functions. This study aimed to investigate the relationship between Cx43 and lipopolysaccharide (LPS)-induced oxidative stress and apoptosis in human umbilical vein endothelial cells (HUVECs). Methods Western blot was performed to determine mitochondrial Cx43 (MtCx43) protein level and phosphorylation (p-MtCx43). Gap19, a selective Cx43 inhibitor, was used to examine the effects of Cx43 on LPS-induced oxidative stress and apoptosis in HUVECs. Expression of regulatory genes associated with oxidative stress was examined by quantitative polymerase chain reaction (qPCR) and Western blot. Apoptosis was assessed by flow cytometry. Results LPS stimulation resulted in increased levels of MtCx43 and p-MtCx43. Interestingly, Gap19 antagonized the upregulation of glutathione S-transferase Zeta 1 (GSTZ1) and cytochrome b alpha beta (CYBB), and the downregulation of antioxidant 1 (ATOX1), glutathione synthetase (GSS) and heme oxygenase 1 (HMOX1) induced by LPS or Cx43 overexpression. Moreover, the increased production of reactive oxygen species (ROS) and apoptosis elicited by LPS or Cx43 overexpression were reduced following treatment with Gap19. Conclusions Selective inhibition of Cx43 hemichannels protects HUVECs from LPS-induced apoptosis and this may be via a reduction in oxidative stress production.