CDK抑制剂对糖尿病大鼠视网膜Müller细胞胶质增殖及新生血管形成的抑制作用

Inhibitory effects of CDK inhibitors on the glial proliferation of retinal Müller cells and retinal neovascularization in diabetic rats

目的探讨CDK抑制剂对糖尿病大鼠视网膜Müller细胞胶质增殖及新生血管形成的影响。方法雄性SD大鼠30只,随机分成对照组、糖尿病组、治疗组(CDK抑制剂SCH727965),每组各10只。后两组大鼠采用单次腹腔注射55 mg·kg^-1链脲佐菌素(streptozotocin,STZ)的方法诱导糖尿病模型。模型诱导成功后,治疗组玻璃体内注射SCH727965 8μL(3 nmol·L^-1)。12周后,免疫荧光检测三组大鼠视网膜胶质纤维酸性蛋白质(glial fibrillary acidic protein,GFAP)、血管内皮生长因子(vascular endothelial growth factor,VEGF)表达,免疫组织化学检测视网膜色素上皮衍生因子(pigment epithelium-derived factor,PEDF)表达,Western blot检测GFAP、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)、PEDF蛋白相对表达量。结果与对照组GFAP (100.00±0.00)%、VEGF(100.00±0.00)%、PEDF(38.26±0.52)%、PCNA(9.34±0.47)%表达相比,糖尿病组GFAP(168.24±2.72)%、VEGF(156.79±1.75)%、PCNA(16.11±0.34)%表达均明显增加,PEDF(23.72±0.71)%表达明显降低(均为P<0.01);而与糖尿病组相比,治疗组GFAP(124.37±3.01)%、VEGF(118.36±1.98)%、PCNA(12.05±0.67)%表达均明显降低,PEDF(8.22±0.36)%表达明显增加(均为P<0.05)。结论 SCH727965可下调大鼠糖尿病状态下视网膜GFAP、VEGF、PCNA表达,上调PEDF表达,进而抑制Müller细胞增殖及新生血管形成。

Objective To investigate the inhibitory effects of CDK inhibitors on the glial proliferation of retinal Müller cells and retinal neovascularization in diabetic rats.Methods Thirty male SD rats were randomly divided into control group,diabetic group and treatment group(CDK inhibitor SCH727965),10 in each group.The diabetic model was induced by single intraperitoneal(IP) injection of streptozotocin(STZ)(55 mg·kg^-1) in the latter two groups.After successful model induction,the treatment group was given intravitreal injection of 8 μL(3 nmol·L^-1) SCH727965.After 12 weeks,the protein expressions of(glial fibrillary acidic protein,GFAP) and vascular endothelial growth factor,VEGF) in retina of rats in each group were detected by immunofluorescence,and the expressions of pigment epithelium-derived factor(PEDF) was detected by immunohistochemistry,and the relative expressions of GFAP,proliferating cell nuclear antigen(PCNA) and PEDF were detected by Western blot.Results Compared with the expression of GFAP(100.00±0.00)%,VEGF(100.00±0.00)%,PEDF(38.26±0.52)%,PCNA(9.34±0.47)% in the control group,the expression levels of GFAP [(168.24±2.72)%],VEGF [(156.79±1.75)%],PCNA [(16.11±0.34)%] were significantly increased and PEDF level [(8.22 +0.36)%] was significantly decreased in the diabetic group(all P<0.01).Compared with diabetes group,the expression levels of GFAP [(124.37±3.01)%],VEGF [(118.36±1.98)%],PCNA [(12.05±0.67)%] were decreased significantly and the expression level of PEDF [(8.22±0.36)%] was increased significantly in the treatment group(all P<0.05).Conclusion SCH727965 can down-regulate the expression of GFAP,VEGF and PCNA and up-regulate the expression of PEDF in diabetic rats,and then inhibit the retinal neovascularization and glial proliferation of retinal Müller cells.