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Ang-1 on high glucose in the culture of RF/6A


Of: left, Feng Wang Yijing Fu Chuang based economy and
【Abstract】 Objective


angiopoietin -1 (angiopoietin-1, Ang-1) on high glucose in cultured monkey choroid - retinal endothelial cells (RF/6A) in rats. Method of training RF/6A divided into 3 groups: control group (DMEM medium containing 25 mmol / L glucose),[link widoczny dla zalogowanych], high glucose (DMEM medium containing 40 mmol / L glucose), Ang-high glucose (DMEM medium containing 40 mmol / L glucose +0.25 mg / L Ang-1). Using trypan blue staining and MTT colorimetric assay in vitro survival and cell viability of various RF/6A, using western-blotting method to detect the suivivin expression. Results 1 d when the cells were no differences in survival and viability. 3,5,7 d when the high glucose group and the Ang-high glucose cell survival and cell viability decreased compared with the normal group (P angiopoietin -1; high glucose; survivin expression 【Abstract】 Objective To explore the damage of high glucose on rhesus macaque choroids-retinal endothelial cell (RF/6A) and also evaluate the protective effect and mechanisms of Angiopoietin-1 (Ang-1) on RF/6A in this environment.Methods RF/6A were divided into 3 groups to be cultured: (1) control group (DMEM containing 25 mmol / L glucose) ; (2) high glucose group (DMEM containing 40 mmol / L glucose); (3) Ang-high glucose group (DMEM containing both 40 mmol / L glucose and 0.25 mg / L Ang-1). Each group were evaluated by measurement of MTT, trypan blue dye exclusion method at the 1 st, 3rd,[link widoczny dla zalogowanych], 5th and 7th day respectively, and evaluated by western-blotting at the 5th day to determine the expression of survivin.Results At the 3rd, 5th and 7th day, lower cell proliferation and livability in high glucose group and Ang-high glucose group were showed, compared with control group (P <0.05), the same tendency was observed in high glucose group compared with Ang-high glucose group (P <0.05). The western -blotting showed the expression of survivn in Ang-high glucose group was significantly higher than other two groups (P <0.05). Conclusion High glucose can decrease the proliferation and livability of RF/6A while Ang-1 limits these damages in high glucose environment may by way of up regulating the expression of survivin. 【Key words】 Angiopoietin-1; High glucose; Expression of surviving diabetic retinopathy (diabetic retinopathy, DR) is the most common and most serious diabetes One of the complications that can lead to decreased visual acuity or even blindness. The disease pathogenesis is not fully understood, but endothelial cells (endothelial cell, EC) and pericytes (pericyte) accelerated apoptosis in the development of DR has played an important role. Angiopoietin -1 (angiopoietin-1, Ang-1) involved in the regulation of vascular maturation, maintenance of EC stability,[link widoczny dla zalogowanych], is to determine the regression of atrophy or vascular proliferation and leakage of the important factors [1-4]. Although Ang-1 on the physiological effects of more, but Ang-1 in high glucose in the study of the effects on EC has been reported rarely. In this study, that EC RF/6A this preliminary study carried out in order to apply for the Ang-1 to provide a theoretical basis for clinical treatment of DR. 1 Materials and methods 1.1 Materials and equipment monkey choroid - retinal endothelial cells (Shanghai Institute of Cell Bank), DMEM high glucose medium (Gibco Company),[link widoczny dla zalogowanych], survivin antibody (Santa Cruz Company) Trypan Blue (sigma company), MTT (sigma company), the Reader (Beijing Planck companies) Ultrasonic Cell Crusher (Ningbo Institute of Chicago Section devices, 92 - Ⅱ type), Kodak electrophoresis image analysis system (Kodak Company, EDAS290 type). 1.2 1.2.1 Experimental Methods Experimental groups will RF/6A in DMEM high glucose complete medium (containing sodium pyruvate, 2 mmol / L L-glutamine, 15% fetal bovine serum, 100 U / ml penicillin, 0.1 mg / ml streptomycin) at 37 ℃, 5% CO 2, humidified incubator in culture, when cells covered 90% of cells after the bottom,[link widoczny dla zalogowanych], with 0.25% trypsin / 0.02% EDTA digestion , in accordance with the passaged 1:2 1:4 training groups: control group (DMEM medium containing 25 mmol / L glucose), high glucose (DMEM medium containing 30 mmol / L glucose), Ang-high glucose (DMEM medium containing 30 mmol / L glucose +0.25 mg / L Ang-1). 1.2.2 Trypan blue cell viability test to adjust the cell concentration to 1 × 10 4 / inoculated in the density of holes 96, each well 200 μl. 4 h before testing into the corresponding serum-free medium, each well add MTT (5 mg / ml) 20 μl, 37 ℃, after 4 h incubation medium was discarded hole, add 150 μl dimethyl sulfoxide, the oscillation 15 min to fully dissolve crystals. Zero with an empty hole, 570 nm wavelength detection, 630 nm wavelength calibration, the instrument was determined by ELISA absorbance value of each hole (A value), according to a formula the proportion of experimental group of cell survival: cell viability = test group A value / control group A values × 100%. More articles related to topics：


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