Zhang Wei, Han Qi, Chen Song, Yan Hua
Department of Ophthalmology, Tianjin Medical University General Hospital, Tianjin 300052, China.
Zhonghua Yan Ke Za Zhi. 2012 Nov;48(11):1015-20.
To observe the effect of simvastatin on the amount and function of endothelial progenitor cells from bone marrow in diabetic rats.
Experimental study. Twenty male Wistar rats were induced with streptozotocin (STZ) injection for the establishment of diabetic retinopathy model. Mononuclear cells were collected by density gradient centrifugation from the bone marrow of rats. The isolated cells were cultivated in dishes coated with fibronectin. Cultured cells were divided into normal control (CON) group and the intervention group. Cells in the CON group were cultured with regular culture medium and the intervention group was treated with 0.01, 0.1, 1 and 10 µmol/L of simvastatin. Immuno-fluorescence staining and flow cytometry were used to identify EPC. The colony number of EPC was assayed by CFU counting. Proliferation, migration and adhesion function of EPC were assayed by MTT chromatometry, modified Boyden chamber assay and adhesion activity assay, respectively. All eyeballs were examined by histopathological examination stained by hematoxylin and eosin (HE) and electron microscopic examination. The amount, proliferation, migration and adhesion function of EPC were analyzed by one-way ANOVA and LSD-t methods.
The cell clusters number of bone marrow-derived EPC was significantly increased in intervention groups [(16.5 ± 1.6), (19.0 ± 2.3), (21.9 ± 2.0), (13.9 ± 2.4) n/200 fields in cultures treated with 0.01, 0.1, 1 and 10 µmol/L of simvastatin] compared with that in the CON group [(14.0 ± 2.4) n/200 fields]. The proliferation ability of EPC was increased in intervention groups (0.105 ± 0.014, 0.133 ± 0.024, 0.202 ± 0.039 and 0.068 ± 0.011) compared with that in the CON group (0.072 ± 0.011). The migration ability of EPC was increased in intervention groups [(10.5 ± 1.6), (12.9 ± 2.2), (15.9 ± 2.4), (9.4 ± 1.4) cells/200 fields] compared with that in the CON group [(8.9 ± 1.2) cells/200 fields]. The adhesion ability of EPC was increased in intervention groups [(10.6 ± 1.9), (15.1 ± 2.7), (19.0 ± 3.9), (7.9 ± 1.2) cells/200 fields] compared with that in the CON group [(7.5 ± 1.2) cells/200 fields]. The thickness of retina was reduced and retinal cells became disorganized in diabetic rats. Transmission electron microscopy showed capillary lumen stenosis and retinal microaneurysms with severe local tissue ischemia such as vacuolar degeneration perivascular tissue.
Simvastatin can enhance the amount and function of EPC from bone marrow in diabetic rats in a dose-dependent manner.
观察辛伐他汀对糖尿病大鼠骨髓内皮祖细胞数量及功能的影响。
实验研究。20只雄性Wistar大鼠经链脲佐菌素（STZ）注射诱导建立糖尿病视网膜病变模型。采用密度梯度离心法从大鼠骨髓中收集单个核细胞。将分离的细胞接种于包被纤连蛋白的培养皿中。培养的细胞分为正常对照组（CON）和干预组。CON组细胞用常规培养基培养，干预组分别用0.01、0.1、1和10 μmol/L辛伐他汀处理。采用免疫荧光染色和流式细胞术鉴定内皮祖细胞（EPC）。通过集落形成单位（CFU）计数检测EPC的集落数。分别采用MTT比色法、改良Boyden小室法和黏附活性测定法检测EPC的增殖、迁移和黏附功能。所有眼球均进行苏木精-伊红（HE）染色组织病理学检查和电子显微镜检查。采用单因素方差分析和LSD-t检验分析EPC的数量、增殖、迁移和黏附功能。
与CON组[（14.0±2.4）个/200视野]相比，干预组[用0.01、0.1、1和10 μmol/L辛伐他汀处理的培养物中分别为（16.5±1.6）、（19.0±2.3）、（21.9±2.0）、（13.9±2.4）个/200视野]骨髓来源EPC的细胞簇数量显著增加。与CON组（0.072±0.011）相比，干预组EPC的增殖能力增强（分别为0.105±0.014、0.133±0.024、0.202±0.039和0.068±0.011）。与CON组[（8.9±1.2）个/200视野]相比，干预组EPC的迁移能力增强[（10.5±1.6）、（12.9±2.2）、（15.9±2.4）、（9.4±1.4）个/200视野]。与CON组[（7.5±1.2）个/200视野]相比，干预组EPC的黏附能力增强[（10.6±1.9）、（15.1±2.7）、（19.0±3.9）、（7.9±1.2）个/200视野]。糖尿病大鼠视网膜厚度变薄，视网膜细胞排列紊乱。透射电子显微镜显示毛细血管腔狭窄和视网膜微动脉瘤，伴有严重的局部组织缺血，如血管周围组织空泡变性。
辛伐他汀可剂量依赖性地增强糖尿病大鼠骨髓EPC的数量及功能。
