梁小玲 1 , 魏丽清 2 , 周焕娇 2 , 孙刚 2 , 杨诚 3 , 张熙芳 2 , 杨璐 3 , 林佩蓉 3
梁小玲, Email:
Procollagen proline dioxygenase/antagonists and inhibitors; Vascular endothelial growth factors; Animal experimentation
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Objective  To observe the influence of prolyl hydroxylase 2 (PHD2) expression on endothelial barrier dysfunction induced by high glucose in human retinal vascular endothelial cells (HRECs). Methods  The HRECs were treated by different culture medium with various glucose concentrations (5 mmol/L glucose, 5 mmol/L glucose +25 mmol/L mannitol, 30 mmol/L glucose) as normal control group, mannitol control group and high glucose group, respectively. After the cells cultured for 24 and 48 hours, the protein levels of PHD2, hypoxia-inducible factor-1 alpha; (HIF-1 alpha;) and occludin was detected by Western blot; the expression of vascular endothelial growth factor (VEGF) in the supernatant was determined by enzymelinked immuno sorbent assay (ELISA); the transcription levels of PHD2, HIF-1 alpha;, VEGF and occludin were determined by the reversetranscription polymerase chain reaction (RT-PCR); the paracellular permeability between endotheliums was detected by 7 times;104 molecular weight FITCdextran. Results  Compared with normal control group, the protein level of PHD2 in mannitol control group and high glucose group firstly decreased and then increased, the protein level of HIF-1 alpha; increased while that of occludin decreased; the secretion of VEGF increased in high glucose group but not in mannitol control group (PHD2：F=7.618, 8.627;P＜0.05. HIF-1 alpha;： chi;2=7.692, 7.652;P＜0.05. occludin：F=23.23, 7.317;P＜0.05. VEGF：F=10.768, 4.562; P＜0.05). Compared with normal control group, the mRNA levels of PHD2, HIF-1 alpha;, VEGF and occludin in mannitol control group and high glucose group increased (PHD2：F=5.69, 14.27;P＜0.05. HIF-1 alpha;：F=6.07, 10.47;P＜0.05. VEGF：F=12.31, 9.14;P＜0.05. occludin：F=8.77, 8.00;P＜0.05). Compared with normal control group, the paracellular permeability of mannitol control group and high glucose group increased ( chi;2=20.57,F=56.09;P＜0.05). Conclusions  High glucose induced altered expression of PHD2 which might play an important role in endothelial barrier dysfunction. The mechanism might be associated with HIF-1 alpha; and VEGF.

Citation： 梁小玲,魏丽清,周焕娇,孙刚,杨诚,张熙芳,杨璐,林佩蓉. Influence of prolyl hydroxylase 2 expression on endothelial barrier dysfunction induced by high glucose in human retinal vascular endothelial cells. Chinese Journal of Ocular Fundus Diseases, 2013, 29(1): 35-39. doi: Copy