Objective To investigate the effects of the gene expression of carbohydrate response element binding protein(ChREBP)on human retinal microvascular endothelial cell(hRMEC)injury and angiogenesis under high-glucose conditions.Methods The hRMECs after 4-6 passages were added to a serum-free medium and cultured for 6 hours.Then,these hRMECs were treated by a normal concentration of glucose(5.6 mmol·L-1),a normal concentration of glu-cose+mannitol(24.4 mmol·L-1),a high concentration of glucose(30.0 mmol·L-1),blank siRNA+a high concentra-tion of glucose,and ChREBP siRNA+a high concentration of glucose,which were designed as the control group,hyperos-motic group,high-glucose alone group,blank siRNA group,and ChREBP knockdown group,respectively.Subsequently,these hRMECs were transfected with Lipofectamine RNAiMAX for 24 hours.Western blot was used to analyze the expression of ChREBP in hRMECs under high-glucose conditions.The terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)assay,cell scratch assay,Transwell migration assay,and tube formation assay were conducted to analyze the apoptosis and migration of these cells and capillary angiogenesis.Normal ChREBP-expressing wild-type mice and ChREBP-knockout mutant mice were selected to construct corresponding animal models,and the number of retinal angiogenic sprouts was counted under a microscope.Results The relative protein expression level of ChREBP in hRMECs after 30 minutes of high-glucose stimulation was significantly higher than that after 15 minutes,1 hour,2 hours,and 3 hours of stimulation(all P＜0.05).Meanwhile,the relative protein expression level of TXNIP in hRMECs after 2 hours of high-glu-cose stimulation was significantly higher than that after 15 minutes,30 minutes,1 hour,and 3 hours of stimulation(all P＜0.05).Under high-glucose conditions,the relative protein expression level of TXNIP and NLRP3 in hRMECs of the ChREBP knockdown group was significantly lower than that of the blank siRNA group and the high-glucose alone group(all P＜0.05).The results of TUNEL assay,cell scratch assay,and capillary tube formation assay showed that the relative apopto-sis rate,relative cell coverage area percentage,and relative migrating cell number percentage of the ChREBP knockdown group were significantly lower than those of the blank siRNA group and the high-glucose alone group,whereas the number of ring structures was significantly lower than that of the blank siRNA group,the high-glucose alone group,and the hyper-osmotic group(all P＜0.05).The relative expression level of p-AKT/AKT and p-mTOR/mTOR in the ChREBP knockdown group was significantly lower than that in the blank siRNA group and the high glucose alone group(all P＜0.05).The num-ber of retinal angiogenic sprouts in ChREBP-knockout mutant mice was(22.81±4.03),which was significantly lower than that of normal ChREBP-expressing wild-type mice(64.35±11.69)(P＜0.05).Conclusion ChREBP can affect the oc-currence and development of diabetic retinopathy by regulating the apoptosis,migration,and tubulogenesis of hRMECs and the development of retinal neovascularization under high-glucose conditions,and hence it can be regarded as a potential target for the treatment of this disease.

Objective To investigate the effects of the gene expression of carbohydrate response element binding protein(ChREBP)on human retinal microvascular endothelial cell(hRMEC)injury and angiogenesis under high-glucose conditions.Methods The hRMECs after 4-6 passages were added to a serum-free medium and cultured for 6 hours.Then,these hRMECs were treated by a normal concentration of glucose(5.6 mmol·L-1),a normal concentration of glu-cose+mannitol(24.4 mmol·L-1),a high concentration of glucose(30.0 mmol·L-1),blank siRNA+a high concentra-tion of glucose,and ChREBP siRNA+a high concentration of glucose,which were designed as the control group,hyperos-motic group,high-glucose alone group,blank siRNA group,and ChREBP knockdown group,respectively.Subsequently,these hRMECs were transfected with Lipofectamine RNAiMAX for 24 hours.Western blot was used to analyze the expression of ChREBP in hRMECs under high-glucose conditions.The terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)assay,cell scratch assay,Transwell migration assay,and tube formation assay were conducted to analyze the apoptosis and migration of these cells and capillary angiogenesis.Normal ChREBP-expressing wild-type mice and ChREBP-knockout mutant mice were selected to construct corresponding animal models,and the number of retinal angiogenic sprouts was counted under a microscope.Results The relative protein expression level of ChREBP in hRMECs after 30 minutes of high-glucose stimulation was significantly higher than that after 15 minutes,1 hour,2 hours,and 3 hours of stimulation(all P＜0.05).Meanwhile,the relative protein expression level of TXNIP in hRMECs after 2 hours of high-glu-cose stimulation was significantly higher than that after 15 minutes,30 minutes,1 hour,and 3 hours of stimulation(all P＜0.05).Under high-glucose conditions,the relative protein expression level of TXNIP and NLRP3 in hRMECs of the ChREBP knockdown group was significantly lower than that of the blank siRNA group and the high-glucose alone group(all P＜0.05).The results of TUNEL assay,cell scratch assay,and capillary tube formation assay showed that the relative apopto-sis rate,relative cell coverage area percentage,and relative migrating cell number percentage of the ChREBP knockdown group were significantly lower than those of the blank siRNA group and the high-glucose alone group,whereas the number of ring structures was significantly lower than that of the blank siRNA group,the high-glucose alone group,and the hyper-osmotic group(all P＜0.05).The relative expression level of p-AKT/AKT and p-mTOR/mTOR in the ChREBP knockdown group was significantly lower than that in the blank siRNA group and the high glucose alone group(all P＜0.05).The num-ber of retinal angiogenic sprouts in ChREBP-knockout mutant mice was(22.81±4.03),which was significantly lower than that of normal ChREBP-expressing wild-type mice(64.35±11.69)(P＜0.05).Conclusion ChREBP can affect the oc-currence and development of diabetic retinopathy by regulating the apoptosis,migration,and tubulogenesis of hRMECs and the development of retinal neovascularization under high-glucose conditions,and hence it can be regarded as a potential target for the treatment of this disease.