Hyperglycemia-induced reactive oxygen species toxicity to endothelial cells is dependent on paracrine mediators.
Hyperglycemia-induced reactive oxygen species toxicity to endothelial cells is dependent on paracrine mediators.
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作者：
Busik JV;Mohr S;Grant MB
本研究确定了高糖暴露和细胞因子治疗对人视网膜内皮细胞（HREC）中活性氧（ROS）产生以及炎症和凋亡途径激活的影响。研究设计和方法-在高血糖条件下，测量HREC、人视网膜色素上皮细胞（HRPE）和人Müller细胞（HMCs）的葡萄糖消耗，并与细胞因子治疗进行比较。用5-（和-6）-氯甲基-2 ′，7 ′-二氯二氢荧光素二乙酸酯（CM-H2 DCFDA）、自旋捕集电子顺磁共振和MitoTracker Red染色检测高糖和细胞因子处理后HREC中ROS的产生。不同的信号级联的激活，包括有丝分裂原活化蛋白激酶途径，酪氨酸磷酸化途径，和细胞凋亡的高糖和细胞因子在HREC中，进行了测定。与HRPE和HMCs相反，HRTS-HRECs不会响应于葡萄糖浓度的增加而增加葡萄糖消耗。与正常葡萄糖（5 mmol/l）相比，HREC暴露于25 mmol/l葡萄糖不会刺激内源性ROS的产生、核因子-κB（NF-κB）、细胞外信号相关激酶（ERK）、p38和Jun NH 2-末端激酶（JNK）的激活、酪氨酸磷酸化、白细胞介素（IL）-1β或肿瘤坏死因子-α（TNF-α）的产生，并且仅轻微影响凋亡细胞死亡途径。与此形成鲜明对比的是，HREC暴露于促炎细胞因子IL-1β或TNF-α增加了葡萄糖消耗、线粒体超氧化物产生、ERK和JNK磷酸化、酪氨酸磷酸化、NF-κB活化和caspase活化。结论：我们的体外实验结果表明，HREC对细胞因子而不是高糖有反应，这表明体内糖尿病相关的视网膜内皮损伤可能是由于葡萄糖诱导的其他视网膜细胞的细胞因子释放，而不是高糖的直接影响。
OBJECTIVE—This study determined the effects of high glucose exposure and cytokine treatment on generation of reactive oxygen species (ROS) and activation of inflammatory and apoptotic pathways in human retinal endothelial cells (HRECs). RESEARCH DESIGN AND METHODS—Glucose consumption of HRECs, human retinal pigment epithelial cells (HRPEs), and human Müller cells (HMCs) under elevated glucose conditions was measured and compared with cytokine treatment. Production of ROS in HRECs was examined using 5-(and-6)-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate (CM-H2DCFDA), spin-trap electron paramagnetic resonance, and MitoTracker Red staining after high glucose and cytokine treatment. The activation of different signaling cascades, including the mitogen-activated protein kinase pathways, tyrosine phosphorylation pathways, and apoptosis by high glucose and cytokines in HRECs, was determined. RESULTS—HRECs, in contrast to HRPEs and HMCs, did not increase glucose consumption in response to increasing glucose concentrations. Exposure of HRECs to 25 mmol/l glucose did not stimulate endogenous ROS production, activation of nuclear factor-κB (NF-κB), extracellular signal–related kinase (ERK), p38 and Jun NH2-terminal kinase (JNK), tyrosine phosphorylation, interleukin (IL)-1β, or tumor necrosis factor-α (TNF-α) production and only slightly affected apoptotic cell death pathways compared with normal glucose (5 mmol/l). In marked contrast, exposure of HRECs to proinflammatory cytokines IL-1β or TNF-α increased glucose consumption, mitochondrial superoxide production, ERK and JNK phosphorylation, tyrosine phosphorylation, NF-κB activation, and caspase activation. CONCLUSIONS—Our in vitro results indicate that HRECs respond to cytokines rather than high glucose, suggesting that in vivo diabetes–related endothelial injury in the retina may be due to glucose-induced cytokine release by other retinal cells and not a direct effect of high glucose.
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影响因子：
1.8
作者：
Distler, C;Dreher, Z
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