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视网膜Mnüer细胞条件性基因敲除血管内皮生长因子对氧诱导视网膜病变小鼠的影响被引量:5
2017年
目的观察视网膜Mnüer细胞条件性基因敲除血管内皮生长因子(VEGF)对氧诱导视网膜病变(OIR)小鼠的影响。方法应用Cre-Loxp重组酶技术建立Mnüer细胞条件性基因敲除VEGF小鼠。Cre阳性为敲除VEGF(CKO)小鼠,Cre阴性为未敲除VEGF(NKO)小鼠。CK0小鼠(CKO组)、NK0小鼠(NKO组)各取20只建立OIR模型,观察两组小鼠在建模过程中的死亡率。小鼠17日龄时,行荧光血管造影视网膜铺片观察小鼠视网膜血管形态,计算无血管区面积占全视网膜面积的百分比;行视网膜石蜡切片苏木精伊红染色计数突破内界膜的血管内皮细胞核数;免疫荧光组织化学染色观察小鼠视网膜中缺氧诱导因子-1α(HIF-1α)的表达。结果OIR建模过程中,CKO组、NKO组小鼠总死亡率分别为65.00%、30.00%;两组小鼠总死亡率比较,差异有统计学意义(Х^2=4.912,P=0.027)。CKO组小鼠视网膜正常血管化推迟,可见大片无血管区,新生血管丛少见,整个视网膜单薄、无层次感;NKO组小鼠视网膜正常血管网状结构可见范围较大,血管密度较高,新生血管丛多见,荧光素渗漏较明显。CKO组、NKO组小鼠视网膜无血管区面积占全视网膜面积的百分比分别为(28.314-11.15)%、(16.82±7.23)%;两者比较,差异有统计学意义(t=2.734,P=0.014)。CKO组、NKO组小鼠平均每张切片突破内界膜的血管内皮细胞核数分别为(26.10±6.37)、(28.80±7.59)个;两者比较,差异有统计学意义(t-=2.437,P=-0.016)。CKO组、NKO组小鼠视网膜均可见HIF一1a呈阳性表达,主要位于神经节细胞层和光感受器细胞层;CKO组小鼠视网膜HIF-1α阳性表达强于NKO组。结论Mnüer细胞条件性基因敲除VEGF明显减弱新生小鼠在OIR环境中的生存能力,抑制部分视网膜新生血管的同时推迟了视网膜正常血管化。
金姬卢一冯佳任艳红杨莉丽顾伟忠
关键词:小神经胶质细胞血管内皮生长因子类小鼠缺氧诱导
Anti-inflammatory effects of a synthetic peptide derived from pigment epithelium-derived factor on H2O2-induced corneal injury in vitro被引量:7
2014年
Background The common pathological characteristics of corneal injury include inflammatory factors activation, vascular endothelial cells or inflammatory cells infiltration into lesions, corneal edema, corneal neovascularization (CNV), and scar formation. PEDF-34 is the functional fragment of pigment epithelium-derived factor (PEDF) that has anti-angiogenic and anti-inflammatory properties and contains an N-terminal 34-amino acid peptide. This study was to investigate the anti- inflammatory effects of PEDF-34 on H202-induced corneal injury in vitro. Methods After cultured in H202 (0.1 mmol/L) for 2 hours, human corneal fibroblasts (HCFs) and human umbilical vein endothelial cells (HUVECs) were treated with PEDF-34-nanoparticles (NPs) at different concentrations (0.1, 0.5, 1.0, 2.0 μg/ml) or 2.0 μg/ml controI-NPs for 24 hours. The viable cells were quantified using the MTT assay. Western blotting or ELISA analysis was performed for measuring the human vascular endothelial growth factor (VEGF) and intercellular adhesion molecule-1 (ICAM-1) expression of both HCFs and HUVECs. VEGF and nuclear factor KB (NF-KB) mRNA levels of HCFs were semi-quantified by RT-PCR. Results The survival rates of HCFs or HUVECs stimulated by H202 did not decrease significantly (P 〉0.05) compared to those in the normal conditions. As compared to controI-NP group, PEDF-34-NPs had dose-dependent inhibitive effect on HUVECs with the MTT assay, but not HCFs. Western blotting analysis showed that the VEGF and ICAM-1 levels in the HCFs and HUVECs stimulated by H202 were significantly higher than those in the normal conditions, which were decreased dramatically in those treated with PEDF-34-NPs. RT-PCR analysis revealed that the VEGF mRNA and NF-KB mRNA levels increased in H202-stimulated HCFs, while both of them decreased in PEDF-34-NP groups dose dependently. Conclusions PEDF-34-NPs may play an important role in regulating the NF-kB pathway, inhibiting inflammatory activity. PEDF-34-NPs may be a po
Lu YiFeng JiaYang LiliTang HongfengJin JiXu Xun
关键词:ANTI-INFLAMMATION
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