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AIM: To investigate the influence of hyperglycemia on the severity of choroidal neovascularization(CNV),especially the involvement of bone marrow-derived cells(BMCs) and underlying mechanisms.·METHODS: BMCs from firefly luciferase(Fluc)/green fluorescent protein(GFP) double transgenic mice were transplanted into C57BL/6J wide-type mice. The recipient mice were injected intraperitoneally with streptozotocin(STZ) daily for 5 consecutive days to induce diabetes mellitus(DM), followed by CNV laser photocoagulation.The BMCs recruitment in CNV exposed to hyperglycemia was firstly examined in Fluc/GFP chimeric mice by in vivo optical bioluminescence imaging(BLI) and in vitro Fluc assays. The CNV severity was evaluated by H&E staining and choroidal flatmount. The expression of vascular endothelial growth factor(VEGF) and stromal cell derived factor-1(SDF-1) was detected by Western blot.·RESULTS: BLI showed that the BMCs exerted dynamic effects in CNV model in Fluc/GFP chimeric mice exposed to hyperglycemia. The signal intensity of transplanted Fluc+GFP+BMCs in the DM chimeric mice was significantly higher than that in the control chimeric mice with CNV induction at days 5, 7, 14 and 21(121861.67 ±9948.81 vs 144998.33 ±13787.13 photons/second/cm2/sr for control and DM mice, P5d<0.05; 178791.67±30350.8 vs240166.67 ±22605.3, P7d<0.05; 124176.67 ±16253.52 vs196376.67 ±18556.79, P14d<0.05; 97951.60 ±10343.09 vs119510.00 ±14383.76, P21d<0.05), which was consistent with in vitro Fluc assay at day 7 [relative light units of Fluc(RLU1)], 215.00±52.05 vs 707.33±88.65, P <0.05; RLU1/relative light units of renilla luciferase(RLU2), 0.90 ±0.17 vs 1.83 ±0.17, P <0.05]. The CNVs in the DM mice were wider than those in the control group at days 5, 7, 14 and21(147.83±17.36 vs 220.33±20.17 μm, P5d<0.05; 212.17 ±24.63 vs 326.83 ±19.49, P7d<0.05; 163.17 ±18.24 vs265.17 ±20.55, P14d<0.05; 132.00 ±10.88 vs 205.33 ±12.98,P21d<0.05). The average area of CNV in the DM group was larger at 7d(20688.67±3644.96 vs 32218.00±4132.69 μm2,P <0.05). The expression of VEGF and SDF-1 was enhanced in the DM mice.·CONCLUSION: Hyperglycemia promots the vasculo-genesis of CNV, especially the contribution of BMCs,which might be triggered by VEGF and SDF-1 production.
AIM: To investigate the influence of hyperglycemia on the severity of choroidal neovascularization (CNV), especially the involvement of bone marrow-derived cells (BMCs) and underlying mechanisms. METHODS: BMCs from firefly luciferase (Fluc) / green fluorescent protein ) double transgenic mice were transplanted into C57BL / 6J wide-type mice. The recipient mice were injected intraperitoneally with streptozotocin (STZ) daily for 5 consecutive days to induce diabetes mellitus (DM), followed by CNV laser photocoagulation. The BMCs recruitment in CNV exposed to hyperglycemia was first examined in Fluc / GFP chimeric mice by in vivo optical bioluminescence imaging (BLI) and in vitro Fluc assays. The CNV severity was evaluated by H & E staining and choroidal flatmount. The expression of vascular endothelial growth factor (VEGF) and RESULTS: The BMCs exerted dynamic effects in CNV model in Fluc / GFP chimeric mice exposed t o hyperglycemia. The signal intensity of transplanted Fluc + GFP + BMCs in the DM chimeric mice was significantly higher than that in the control chimeric mice with CNV induction at days 5, 7, 14 and 21 (121861.67 ± 9948.81 vs 144998.33 ± 13787.13 photons / second / cm2 / sr for control and DM mice, P5d <0.05; 178791.67 ± 30350.8 vs240166.67 ± 22605.3, P7d <0.05; 124176.67 ± 16253.52 vs196376.67 ± 18556.79, P14d <0.05; 97951.60 ± 10343.09 vs119510.00 ± 14383.76, P21d <0.05), which was consistent with in vitro Fluc assay at day 7 [relative light units of Fluc (RLU1)], 215.00 ± 52.05 vs 707.33 ± 88.65, P <0.05; RLU1 / relative light units of renilla luciferase , 0.90 ± 0.17 vs 1.83 ± 0.17, P <0.05]. The CNVs in the more mice than those in the control group at days 5, 7, 14 and 21 (147.83 ± 17.36 vs 220.33 ± 20.17 μm, P5d <0.05; 212.17 ± 24.63 vs 326.83 ± 19.49, P7d <0.05; 163.17 ± 18.24 vs265.17 ± 20.55, P14d <0.05; 132.00 ± 10.88 vs 205.33 ± 12.98, P21d <0.05). The average area of CNV in the DM group was larger at 7d (20688.67 ± 3644.96 vs 32218.00 ± 4132.69 μm 2, P <0.05). CONCLUSION: Hyperglycemia promotors of the vasculo-genesis of CNV, especially the contribution of BMCs, which might be triggered by VEGF and SDF-1 production.