Protection of GAP-43 gene–modified bone marrow mesenchymal stem cells on retinitis pigmentosa
摘要
To observe the therapeutic effects of growth associated protein-43 (GAP 43) gene modified rat bone marrow mesenchymal stem cells (BMSCs) on experimental retinal degeneration.
MethodsA total of 80 male Royal College of Surgeons (RCS) rats were divided into three groups at postnatal 21 (P21) randomly. A cell suspension of 5 × 104 BMSCs modified with GAP-43 in 2 µl PBS was injected into the subretinal space of BMSCs + GAP-43 group rats, BMSCs group rats received5 × 104 BMSCs in 2 µl PBS and PBS group rats received 2 µl PBS. Photoreceptor necrosis and apoptosis was assessed by HE staining and Terminal-deoxynucleotidyl Transferase Mediated Nick End Labeling (TUNEL) detection. The expression of GAP-43, Glutamine synthetase (GS), rhodopsin (RHO), Caspase-8 and Caspase-9 was analyzed by immunofluorescence, western blot.
ResultsTransplantation of GAP-43-modified BMSCs into the subretinal cavity of RCS rats revealed that: retinal GAP-43 expression was significantly elevated (P < 0.05); Rhodopsin (Rho) expression, a photoreceptor marker, was upregulated 1.5-fold (P < 0.05), and the thickness of the outer nuclear layer increased to 79.78 ± 6.83 μm (control 29.92 ± 4.17 μm, P < 0.01); The apoptosis rate decreased to 41.9% (control 63.8%, P < 0.01), and Caspase-8/9 activation was inhibited; Müller cell proliferation was reduced (GS decreased, P < 0.05). suggesting that GAP-43-modified BMSCs delayed retinal degeneration through anti-apoptosis and pro-photoreceptor survival.
ConclusionsThe results suggest that GAP-43 gene modified BMSCs has the therapeutic effect on the early stage of retinal degeneration, which might make more photoreceptors preserved.
Graphical Abstract