Effect Of Telbivudine On Expression Of Vascular Endothelial Growth Factor In Human Podocyte Cells On Cultured Vascular Endothelial

BackgroundPatients with chronic HBV infection need more long-term treatment and may even need life-long antiviral treatment.Anti HBV nucleoside drugs are all metabolically related to the kidney.It is very important to screen out drugs that may protect the kidneys from patients with chronic hepatitis B,who are at high risk of renal dysfunction or have potential renal dysfunction.Many studies have shown that: telbivudine(telbivudine,L-DT)with renal function in patients with continuous improvement of chronic HBV infection in antiviral effect at the same time,the specific mechanism is not clear.ObjectiveAccording to the different concentrations of L-DT on podocyte(Human Podocyte Cells,HPC)influence the proliferation of L-DT,understanding the influence of HPC face in cell proliferation,and cell proliferation level according to the understanding of the optimal concentration,on vascular endothelial growth factor in cytokine levels(Vascular Endothelial,Growth Factor,VEGF).Effect of m RNA on the expression of.The possible mechanism for the protection of renal function presented by L-DT provides a certain experimental basis.MethodsTaking HPC as the research object,the in vitro cultured cells after digestion and passage,choose the logarithmic experiment with free drug telbivudine(L-DT 0 mmol/L)blank group as control group,and 6 different concentration gradient(L-DT 10,20,40,60,100,150mmol/L)for the experiment in the group,cell density was 5000/ hole in advancedisinfection 96 good Kong Banzhong,volume of 100ul/ hole,24 h culture,the cell contact area of 80% after the suction to the culture medium,after 24 h starvation.Each hole added volume prepared in each experimental group were 24 and 48 concentration,72 h with the inverted microscope to observe cell morphology changes and 4-methyl thiazolyl tetrazolium(methyl thiazolyl tetrazolium MTT)method was used to detect cell viability,to understand the effect of telbivudine on proliferation of HPC.The concentration of half inhibitory drug(half maximal inhibitory concentration,IC50)on the proliferation of HPC was calculated by L-DT.According to the three representative concentration of desired RT-PCR the IC50(L-DT10,50,100mmol/L)compared with the control group in response to telbivudine effects on HPC VEGFmRNA expression.The semi quantitative RT-PCR products were gel electrophoresis imaging,and take GAPDH as a reference,the gray values were detected by using related software contrast ratio represents the relative expression of VEGFmRNA with gray VEGF/ GAPDH value.Foreign language version of the software in this experiment using SPSS 21 statistical data,the mean and standard deviation of measurement data taken by(s))said,compared with group ANOVA(analysis of variance detection Levene test variance homogeneity test),P < 0.05,the difference was statistically significant ResultsChanges in cell morphology with inverted microscope observation can be extended:concentrations in cell shape with the increase of drug concentration and action time gradually showing nuclear enrichment,even showing cytoplasmic vacuoles;MTT cell viability assay: when the drug concentration of 100 mmol/L 24,48 After 72 h,the inhibition rate of cells for(40.85 + 1.37)% and(51.03 + 2.15)% and(76.11 + 4.36)%,the difference was statistically significant between two groups(P < 0.05);L-DT HPC IC50100 mmol/L;there were differences between the different inhibition rate of different concentration and action time of HPC,and statistics there was significant difference(P <0.05).Semi quantitative RT-PCR showed that the expression of VEGFmRNA wasdecreased in the three groups of L-DT10,50 and 100mmol/L compared with the blank control group(P<0.05),and the inhibition effect of 100mmol/L on VEGFmRNA expression was more significant(P < 0.05).Conclusions1.L-DT has a inhibitory effect on the proliferation of HPC,and it is time and dose dependent.2.L-DT downregulated the expression of VEGFmRNA.3.L-DT may be involved in the mechanism of renal protection by inhibiting the expression of VEGFmRNA on the patient's HPC.