|
Objective: In this study, by in vitro induced by high glucose umbilical vein endothelial cells (human umbilical vascular endothelial cells, HUVEC) injury, the establishment of the the HUVEC high sugar damage model to explore salvianolate umbilical vein endothelial cells induced by high glucose The protective effect and mechanism of injury. Methods: 1, in vitro HUVEC high glucose injury model in vitro cultured human umbilical vein endothelial cells (Human YANBIAN UNIVERSITY. Protective effects of the compound Danshen vascular endothelial cells, HUVEC) were randomly divided into normal control group (glucose concentration of 5.5mmol / L), high glucose injury Group 5 (glucose concentration, respectively, for 10mmo1 / L 20mmo1 / L, 30mmo1 / L, 40mmo1 / L, 50mmo1 / L), cells were collected after various incubation times (0h, 24h, 48h, 72h) with the appropriate culture medium, cell viability (OD), cell culture supernatant glutathione - peroxidase (GSH-PX) activity, detection of malondialdehyde (MDA), respectively, with an inverted phase contrast microscope cell morphology changes. 2 salvianolate protective effect of high glucose-induced human umbilical vein endothelial cell injury were divided: (1) normal control group (glucose concentration of 5.5 mmol / L); the the injury group ② high sugar (glucose concentration 30mmo1 / L); (3) low-dose salvianolate group (50mg / L salvianolate 30mmol / L glucose); the the ④ middle dose salvianolate group (100mg / L salvianolate 30mmol / L glucose) ; ⑤ high doses salvianolate group (200mg / L salvianolate 30mmol / L glucose); adding the appropriate culture medium the cells were collected 48 hours later, cell viability, cell culture supernatants malondialdehyde ( MDA), glutathione - peroxidase (GSH-PX) activity, endothelin -1 (ET-1), nitric oxide (NO) secretion measured. Results: 1, high-sugar injury model in vitro HUVEC 1) normal control group (5.5 mmol / L), 10 mmol / L, 20 mmol / L, cell viability values ??(OD) within 72 hours, gradually increased with the incubation time. . But at the same time, the growth of 10 mmol / L, 20 mmol / L high glucose group OD values ??significantly lower than 5.5 mmol / L group, a significant difference (P = 0.003) between groups. 30 mmol / L, 40 mmol / L, 50 mmol / L group culture period of time, with the decreased OD value of the extension of time, the comparison between groups was statistically significant (P = 0.02). 2) with the incubation time, the cells in each group of MDA contents increased. The incubation time point (time 0), compared with the normal control group, the higher the concentration of glucose, MDA content increased more obvious comparison between groups was statistically significant (P = 0.01). 3) with the incubation time, the cells in each group content of GSH-PX showed an upward trend. The incubation time point (time 0), compared with the normal control group, the higher the concentration of glucose, the smaller the rate of growth of the content of GSH-PX comparison between groups was statistically significant (P = 0.02). 4) inverted phase contrast microscope, cultured 48h HUVEC, glucose concentration of 10 mmol / L, 20 mmol / L group cell morphology and normal control group (5.5mmol / L) showed no significant difference; 30 mmol / L, 40 mmol / L, 50 mmol / L group of cell morphology compared with the control group was significantly different 30mmol / L, 40mmol / L cells are deformed cell deformation which 40mmol / L group than 30mmol / L, significantly, but the two groups of cells is still clear boundary, 50mmol / L group The cell deformation obvious cell boundary is not clear, the phenomenon of cell shedding. 2, salvianolate protective effect of human umbilical vein endothelial cell injury induced by high glucose compared with the normal control group, high glucose group HUVEC cell viability (OD value), the content of GSH-PX and NO secretion decreased (P = 0.003, P = 0.02, P = 0.01), compared to the control group with high sugar content of MDA, ET-1 secretion was significantly higher (P = 0.01, P = 0.003);, salvianolate permanganate after the intervention, HUVEC cell viability, the content of GSH-PX and NO secretion rebounded significantly (P = 0.02, P = 0.01 P = 0.003), MDA content of ET-1 secretion was significantly decreased (P = 0.03, P = 0.004). Conclusion: High glucose-induced HUVEC cell viability decreased, increased generation of reactive oxygen species, antioxidant enzyme activity, resulting in HUVEC injury, and with the glucose concentration and duration of action of HUVEC extent of the damage; concentration of glucose 30mmol / L, the role of 48 hours endothelial cell damage induced by high glucose in vitro as salvianolate of the best of the protective effect of the experimental conditions. 2, salvianolate has a protective effect of high glucose-induced in vitro endothelial cell injury, its mechanism of action may be scavenging reactive oxygen species by protecting the mitochondria, to improve the vitality of the antioxidant enzyme system of endothelial cells, inhibit the secretion of ET-l The protective effect of high sugar vascular endothelial cells.
|