The PKCβ/HuR/VEGF pathway in diabetic retinopathy
The PKCβ/HuR/VEGF pathway in diabetic retinopathy
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DOI：
10.1016/j.bcp.2010.06.033
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发表时间：
2010-10-15
影响因子：
5.8
通讯作者：
Pascale, A.
中科院分区：
文献类型：
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作者：
Amadio, M.;Bucolo, C.;Pascale, A.
我们研究了糖尿病相关的PKC β激活是否通过链脲佐菌素（STZ）诱导的糖尿病大鼠视网膜中稳定mRNA的人胚胎致死性异常视力（ELAV）蛋白HuR影响VEGF表达。用链脲佐菌素（STZ）诱导大鼠糖尿病模型。处理视网膜组织以检测PKC β I、PKC β II、VEGF和HuR含量以及HuR磷酸化。免疫沉淀结合RT-PCR被用来评估核蛋白复合物中HuR与VEGF mRNA的结合。通过ANOVA进行统计分析，然后进行适当的事后比较检验。实验性糖尿病后，与假手术组相比，PKC β I和PKC β II水平升高;还存在PKC介导的HuR磷酸化/激活。这些作用通过体内共同给予选择性PKC β抑制剂而减弱。还检测到HuR蛋白与VEGF mRNA之间的特异性结合。PKC β/HuR激活伴随着VEGF蛋白表达的增强，这再次被PKC β抑制剂钝化。这些发现首次证实了实验性糖尿病后视网膜中PKC β/HuR/VEGF通路的激活，并揭示了一种新的潜在药理学靶点，以抵消涉及VEGF失调的病理学，如糖尿病视网膜病变。(C)2010年爱思唯尔公司All rights reserved.
We investigated whether the diabetes-related PKC beta activation affects VEGF expression through the mRNA-stabilizing human embryonic lethal abnormal vision (ELAV) protein, HuR, in the retina of streptozotocin (STZ)-induced diabetic rats. Diabetes was induced in rats by STZ injection. Retinal tissues were processed to detect PKC beta I, PKC beta II, VEGF and HuR contents, as well as HuR phosphorylation. Immunoprecipitation coupled to RT-PCR was employed to evaluate HuR binding to VEGF mRNA in RiboNucleoProteic (RNP) complexes. Statistical analysis was performed by ANOVA followed by an appropriate post hoc comparison test. Following experimental diabetes PKC beta I and PKC beta II levels were increased compared to sham; there was also a PKC-mediated phosphorylation/activation of HuR. These effects were blunted by the in vivo co-administration of a selective PKC beta inhibitor. A specific binding between the HuR protein and the VEGF mRNA was also detected. The PKC beta/HuR activation was accompanied by enhanced VEGF protein expression that was, again, blunted by the PKC beta inhibitor. These findings first demonstrate the activation, in the retina, of the PKC beta/HuR/VEGF pathway following experimental diabetes and disclose a new potential pharmacological target to counteract pathologies implicating VEGF deregulation, such as diabetic retinopathy. (C) 2010 Elsevier Inc. All rights reserved.