Protective effect and mechanism of Krüppel-like factor 7 overexpression on retinal ganglion cells in mice after optic nerve clamp
Objective To explore the protective effect and mechanism of Krüppel-like factor 7(KLF7)overexpression on retinal ganglion cells(RGC)in mice after optic nerve crush(ONC).Methods A total of sixty five 10-week-old C57BL/6J mice were randomly divided into five groups:blank control group(group A),intravitreal injection(IVT)-KLF7 group(group B),IVT-phosphate buffer saline-ONC group(group C),IVT-KLF7-ONC group(group D),and IVT-recombinant adeno-associated virus 2-enhanced green fluorescent protein-ONC group(group E),with 13 mice in each group.On the 7 days after the ONC model,the mice in each group were killed.RGC survival rate was counted by whole retina flat mount and immunofluorescence techniques.KLF7,nerve growth factor(NGF),tyrosine kinase A(TrkA),phosphorylated TrkA(pTrkA),tyrosine kinase B(TrkB)and phosphorylated TrkB(pTrk)were detected by western blot,growth associated protein 43(GAP43),brain-derived neurotrophic factor,B lymphoblastoma-2(Bcl-2),Bcl-2 associated X protein(BAX),Caspase-3 protein relative expression levels.One-way analysis of variance or Kruskal-Wallis test were used for comparison between groups.Results On the 7 days after the ONC model,the density of RGC in the retina of groups A,B,C,D and E were(3 707.4±12.8),(3 582.4±13.3),(1 396.3±16.1),(1 658.3±22.2)and(1 323.6±16.9)/mm2,respectively.Compared with groups C and E,RGC density in group D was significantly increased,and the difference was statistically significant(P=0.028,0.007).Compared with groups A,B,C and E,the relative expression levels of NGF,pTrkA,pTrkB,GAP43 and Bcl-2 proteins in the retina of mice in group D were increased,while the relative expression levels of BAX and Caspase-3 proteins were decreased,with statistical significance(P＜0.01).Conclusion In mouse ONC model,overexpression of KLF7 can improve RGC survival rate,increase the relative expression levels of NGF,pTrkA,pTrkB,GAP43 and Bcl-2 proteins in retina,and decrease the relative expression levels of BAX and Caspase-3 proteins.
Kruppel-like factor 7Optic nerve clampRetinal ganglion cellsNerve growth factorTyrosine kinaseB lymphoblastoma-2B lymphoblastoma-2-associated X protein
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