Effects Of Telbivudine On Expression Of Angiotensin Ⅱ Type 1a Receptor In Glomerular Mesangial Cells Of Rats

BackgroundChronic hepatitis B(CHB)is caused by hepatitis B virus(HBV)infection,which has become a major infectious disease that threatens global human health.At present,the control of HBV-induced liver injury has become our critical task and the ultimate goal,to find effective antiviral drugs has become our ultimate goal of the bridge.Global multicenter studies have shown that patients with mild renal insufficiency with CHB are treated with L-DT for more than 48 weeks,glomerular filtration rate in these patients who were significantly higher than the baseline level,but the specific mechanism of action is not clear.ObjectiveTo investigate whether L-DT has an effect on the proliferation of rat glomerular mesangial cells(RMCs),it is up-regulated or down-regulated the expression of Angiotensin Ⅱ type 1a receptor(AT1aR),which is closely related to glomerular blood flow and filtration function.In order to further study the possible molecular mechanism of L-DT in improving the glomerular filtration rate of patients,,it provides some experimental basis.MethodsAfter digestion and passage in vitro,we used the logarithmic growth phase of RMCs to experiment.The number of RMCs counted by 5 × 104 cells / mL,according to the volume of 100 ul per well containing 5000 cells,then we put the cells evenly into the three 96-well cell culture plates.According to the L-DT drug concentration,the experiment was divided into blank control group(L-DT 0mmol/L)and experimental group(L-DT 1,10,40,60,100,150mmol/L)and RMCs were treated with L-DT for 24,48 and 72 hours.Then,the thiazolyl tetrazolium(MTT)method was used to detect the optical density(OD)of the cells at different time after different drug concentrations,and the half maximal inhibitory concentration(IC50)of L-DT on the proliferation of RMCs was calculated.The expression of AT1 aR mRNA in RMCs was measured by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR)in blank control group and three different concentration group low(10mmol/L),medium(50mmol/L)and high(100mmol/L),which is according to the value of IC50.The RT-PCR product was subjected to agarose gel electrophoresis,and the band gray value was measured according to Image-J software.The relative expression of AT1 aR mRNA was expressed by the ratio of gray value of AT1 aR / β-actin.Data entry SPSS 19.0 statistical software for analysis.The experimental data for the measurement data to mean ± standard(x ± s)deviation,the experimental data variance homogeneity test Levene test method,compared between groups using ANOVA variance analysis,P <0.05 for the difference was statistically significant.ResultsThe RMCs in the L-DT group showed shrinkage with the increase of the drug concentration and the time of action,and the nuclei were contracted and even cytoplasmic vacuoles.MTT method showed that RMCs in the experimental group were treated with different L-DT concentrations(1,10,40,60,100,150 mmol/L),and the L-DT group IR % Were higher than those in the control group(P <0.05).The IR% of RMCs was 50% after 48 h of L-DT at 100 mmol/L.Semi-quantitative RT-PCR results showed that compared with the control group(L-DT 0mmol / L),Semi-quantitative RT-PCR results showed that L-DT low(10mmol / L),medium(50mmol / L),high(100mmol / L)three groups compared with the blank control group were down-regulated RMCs AT1 aR mRNA expression(P <0.05),And the down-regulation of AT1 aR mRNA expression was more significant in high-dose group(P <0.05).Conclusions1.L-DT inhibited the proliferation of RMCs in a time-and concentration-dependent manner.2.L-DT down-regulates the expression of AT1 aR mRNA on RMCs.3.L-DT may be by inhibiting the expression of AT1 aR mRNA on glomerular mesangial cells to improve the level of GFR in patients.