Activation of the VEGF-A/ERK/PLA2 Axis Mediates Early Retinal Endothelial Cell Damage Induced by High Glucose: New Insight from an In Vitro Model of Diabetic Retinopathy.
Activation of the VEGF-A/ERK/PLA2 Axis Mediates Early Retinal Endothelial Cell Damage Induced by High Glucose: New Insight from an In Vitro Model of Diabetic Retinopathy.
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VEGF-A/ERK/PLA 2轴的激活介导高糖诱导的早期视网膜内皮细胞损伤：来自糖尿病视网膜病变体外模型的新见解
DOI：
10.3390/ijms21207528
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发表时间：
2020-10-13
影响因子：
5.6
通讯作者：
Bucolo C
中科院分区：
文献类型：
--
作者：
Giurdanella G;Lupo G;Gennuso F;Conti F;Furno DL;Mannino G;Anfuso CD;Drago F;Salomone S;Bucolo C
高血糖诱导的早期血视网膜屏障（BRB）功能障碍与磷脂酶A2（PLA 2）的促炎活性增加和血管内皮生长因子A（VEGF-A）的上调有关。在这里，我们测试了VEGF-A在高糖（HG）诱导的人视网膜内皮细胞（HREC）损伤中的作用，该损伤由Ca++依赖性（cPLA 2）和Ca++非依赖性（iPLA 2）PLA 2介导。将HREC用正常葡萄糖（5 mM，NG）或高葡萄糖（25 mM，HG）处理48 h，有或没有VEGF捕获剂阿柏西普（Afl，40 μg/mL）、cPLA 2抑制剂花生四烯酸三氟甲基酮（AACOCF 3; 15 μM）、iPLA 2抑制剂溴烯醇内酯（BEL; 5 μM）或VEGF-A（80 ng/mL）。Afl和AACOCF 3均防止HG诱导的损伤（MTT和LDH释放）、血管生成潜能的损害（管形成）和VEGF-A mRNA的表达。此外，Afl抵消了HG诱导的磷酸化ERK和磷酸化cPLA 2的增加（免疫印迹）。HG培养基中的VEGF-A通过上调磷酸化ERK、磷酸化cPLA 2和iPLA 2（分别约为55%、45%和50%）增加葡萄糖毒性;免疫细胞化学证实了这些蛋白质的活化。siRNA敲除cPLA 2可完全防止HG或HG + VEGF-A诱导的细胞损伤，而iPLA 2敲除产生较温和的保护作用。这些数据表明，VEGF-A通过参与ERK 1/2/PLA 2轴激活介导早期葡萄糖诱导的视网膜内皮损伤。
Early blood retinal barrier (BRB) dysfunction induced by hyperglycemia was related to increased pro-inflammatory activity of phospholipase A2 (PLA2) and the upregulation of vascular endothelial growth factor A (VEGF-A). Here, we tested the role of VEGF-A in high glucose (HG)-induced damage of human retinal endothelial cells (HRECs) mediated by Ca++-dependent (cPLA2) and Ca++-independent (iPLA2) PLA2s. HRECs were treated with normal glucose (5 mM, NG) or high glucose (25 mM, HG) for 48 h with or without the VEGF-trap Aflibercept (Afl, 40 µg/mL), the cPLA2 inhibitor arachidonoyl trifluoromethyl ketone (AACOCF3; 15 µM), the iPLA2 inhibitor bromoenol lactone (BEL; 5 µM), or VEGF-A (80 ng/mL). Both Afl and AACOCF3 prevented HG-induced damage (MTT and LDH release), impairment of angiogenic potential (tube-formation), and expression of VEGF-A mRNA. Furthermore, Afl counteracted HG-induced increase of phospho-ERK and phospho-cPLA2 (immunoblot). VEGF-A in HG-medium increased glucose toxicity, through upregulation of phospho-ERK, phospho-cPLA2, and iPLA2 (about 55%, 45%, and 50%, respectively); immunocytochemistry confirmed the activation of these proteins. cPLA2 knockdown by siRNA entirely prevented cell damage induced by HG or by HG plus VEGF-A, while iPLA2 knockdown produced a milder protective effect. These data indicate that VEGF-A mediates the early glucose-induced damage in retinal endothelium through the involvement of ERK1/2/PLA2 axis activation.
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影响因子：
4.5
作者：
Gong Y;Jin X;Wang QS;Wei SH;Hou BK;Li HY;Zhang MN;Li ZH
通讯作者：
Li ZH
影响因子：
4.1
作者：
Boslem, Ebru;MacIntosh, Gemma;Biden, Trevor J.
通讯作者：
Biden, Trevor J.
DOI：
10.1016/s1054-3589(08)61011-x
发表时间：
1995-01-01
期刊：
Advances in pharmacology (San Diego, Calif.)
影响因子：
--
作者：
Glaser, K B
通讯作者：
Glaser, K B
影响因子：
3.3
作者：
Askarova, S.;Yang, X.;Sheng, W.;Sun, G. Y.;Lee, J. C-M
通讯作者：
Lee, J. C-M
影响因子：
3.1
作者：
Anfuso, Carmelina Daniela;Giurdanella, Giovanni;Alberghina, Mario
通讯作者：
Alberghina, Mario