生长相关蛋白43基因修饰脂肪间充质干细胞对视网膜色素变性的修复及保护

Growth-associated protein-43 gene modified adipose-derived mesenchymal stem cells protect against retinitis pigmentosa

背景:研究发现基因治疗和细胞移植治疗视网膜色素变性有效果,而且二者相结合治疗视网膜色素变性效果更为显著,但其作用机制有待深入研究。目的:探讨生长相关蛋白43基因修饰脂肪间充质干细胞对视网膜色素变性大鼠视网膜的保护作用。方法:分离和培养视网膜色素变性大鼠脂肪间充质干细胞,以生长相关蛋白43慢病毒载体转染脂肪间充质干细胞。将60只视网膜色素变性大鼠分3组干预,实验组视网膜下腔注射生长相关蛋白43慢病毒载体转染的脂肪间充质干细胞,对照组注射脂肪间充质干细胞,假手术组注射PBS,注射后3,7,14,21 d,观察眼球组织脂肪间充质干细胞变化及视网膜生长相关蛋白43表达;注射后28 d,观察视网膜Rho蛋白、GS蛋白表达及外核层厚度。结果与结论:(1)实验组脂肪间充质干细胞移植后,随着时间的延长,生长相关蛋白43表达量逐渐升高,不同时间点间两两比较差异均有显著性意义(P<0.05);(2)实验组、对照组Rho蛋白和GS蛋白表达量高于假手术组(P<0.05);实验组Rho蛋白表达量高于对照组(P<0.05),GS蛋白表达量低于对照组(P<0.05);(3)实验组、对照组视网膜外核层厚度高于假手术组(P<0.05),实验组视网膜外核层厚度高于对照组(P<0.05);(4)结果表明,生长相关蛋白43基因修饰的脂肪间充质干细胞,对视网膜色素变性大鼠模型视网膜具有保护作用。

BACKGROUND:Gene therapy and cell transplantation therapy,especially their combined use,have certain therapeutic effects on retinitis pigmentosa.However,little attention has been paid to the combination of gene therapy and cell transplantation in the treatment of retinitis pigmentosa.OBJECTIVE:To investigate the protective effect of adipose-derived mesenchymal stem cells(ADMSCs)modified by growth-associated protein-43(GAP-43)gene on the retina of rats with retinitis pigmentosa.METHODS:ADMSCs from rats with retinitis pigmentosa were isolated and cultured,and then transfected with GAP-43lentiviral vector.Sixty retinitis pigmentosa rats were randomized into experimental group(subretinal injection of GAP-43-transfected ADMSCs),control group(subretinal injection of ADMSCs)and sham operation group(PBS injection).The expressions of GAP-43protein,Rho protein and GS protein in the retina of rats were determined by western blot.The outer nuclear layer thickness of the retina was determined by hematoxylin-eosin staining.RESULTS AND CONCLUSION:After transfected cell transplantation,the expression of GAP-43protein in the retina was gradually increased with time and showed significant differences at different time post-transplantation(P<0.05).The expression levels of Rho protein and GS protein in the retina of experimental and control groups were higher than that in the sham operation group(P<0.05).The expression of Rho protein in the retina of the experimental group was higher than that in the control group(P<0.05).The expression of GS protein in the retina of the experimental group was lower than that in the control group(P<0.05).The thickness of retinal outer nuclear layer was ranked as follows:the experimental group>the control group>the sham group,and there was significant difference between groups(P<0.05).These results show that the ADMSCs modified by GAP-43gene have protective effect on the retina of rats with retinitis pigmentosa.