| Objective: The purpose of this study the effect of expression for NF-κ B, Caspase-3,and8hydroxy deoxyguanosine (8-OHdG) in cultured human umbilical vein endothelialcells (HUVECs) and the intervention of Liraglutide, to further explore that Liraglutidecan truly and effectively protect the injury of vascular endothelial cells, providing a newway of prevention for diabetic vascular complications.Method:HUVECs, cultured with different concentrations of glucose (respectively:5.5,11.1,25,33.3mmol/L)、high glucose (33.3mmol/L) in different concentrations ofLiraglutide (5,10,25,50,100nmol/L) in vitro and respectively incubated for24or48h.with MTT (four methyl thiazolyl tetrazolium) assay for the detection of cellsurvival rate of each experimental group; According to the results, cells were dividedinto5groups for the normal control group (5.5mmol/L glucose), high glucose group(33.3mmol/L), glucose (33.3mmol/L)+low dose Liraglutide group (25nmol/L), glucose(33.3mmol/L)+medium dose Liraglutide group (50nmol/L) and glucose (33.3mmol/L)+high dose of Liraglutide group (100nmol/L), respectively incubated for24or48h,then measureing the expression of HUVECs in determination of each experimentalgroup of NF-κ B mRNA by using RT-PC.,using enzyme linked immunosorbent assay(ELISA) method for determinating of the expression level of Caspase-3、8-OHdG in thesupernatants of the cells in the experimental group, the obtained data were statisticallyanalyzed with using SPSS19.0software.Results:1. MTT method for detecting cell survival rate: Compared with the normal controlgroup (glucose at the concentration of5.5mmol/L),25、33.3mmol/L glucoseconcentration group at different incubation time (24、48h), the reducement extent of theOD values shows statistical significance (P<0.01),The high glucose group(glucoseconcentration is33.3mmol/L) was decreased most obviously (P<0.01), And under acertain effect of high glucose concentration (33.3mmol/L). Lesions were graduallyaggravated with the extension of time (P<0.01); Compared with high glucose group, theOD values are all improved which added different concentrations of Liraglutide group.Comparison of the25,50,100nmol/L Liraglutide groupand high glucose group,there is statistical significance for the OD values increased of all experimental groups atdifferent incubation time (24、48h), Liraglutide, especially the concentration of100nmol/L, having strongest effect.2. Determination of RT-PCR for expression of HUVECs cells in NF-κ B mRNA:HUVECs expression of NF-κ B mRNA level of normal control group is low; for highglucose group, HUVECs NF-κ B mRNA levels increased significantly compared withnormal environment (P<0.01); with the intervention of different concentrations ofLiraru peptide, HUVECs NF-κ B mRNA level significantly reduced than before, whichLiraru peptide (50,100nmol/1) intervention in24、48h,NF-κ B mRNA expression wasstatistically significant (P<0.01). Liraglutide and HUVECs were incubated for24hours,with Liraglutide concentrations (25,50,100nmol/L) increased, the expression of NF-κB mRNA levels were significantly reduced (P<0.01).3. Determination of the expression level of HUVECs cell supernatants of Caspase-3、8-OHdG by ELISA: Compared with the normal control group, HUVECs cellsCaspase-3、8-OHdG secretion of high glucose group increased significantly (P<0.01),Compared with high glucose group, after treated with different concentrations of theLiraglutide, the secretion capacity of HUVECs cells Caspase-3、8-OHdG significantlyreduced, and also in a concentration dependent manner and Liraglutide. Amongthem,there is statistical significance (P<0.01).Conclusion1.High glucose accelerates the injury for the cell chich cultured human umbilical veinendothelial in vitro, and the extent of cell injury in a dose and time dependent.2.Liraglutide reduces the expression of NF-κB mRNA in cells, decreasing the oxidativestress in endothelial cells.3.Liraglutide can reduce the expression cell supernatant of Caspase-3、8-OHdG, anddecrease the apoptosis of endothelial cells which is induced by high glucose andoxidative injured endothelial cells reduced at the same time.4.Liraglutide has the protective effect on the apoptosis of endothelial cells which areinduced by high glucose in vitro,the mechanism may be related to reduced expression of NF-κ B mRNA,Caspase-3,8-OHdG. |
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