Flow cytometry data are expressed as the geometric imply fluorescence intensity or the percentage of stained cells. The Kolmogorov Smirnov test indicated that the variables displayed a standard distribution. The statistical evaluation was performed working with one particular way evaluation of variance. When considerable variations were demon strated by ANOVA, the post hoc Bonferronis test was per formed. The statistical analyses had been performed employing the Prism application. The variations have been regarded signifi cant when p 0. 05. Results Physique and kidney weights, MAP and HR parameters and angiotensin II levels Initial body weight was statistically similar amongst the groups. By the finish of the experiments, only the 2K1C group displayed lowered physique weight.
Twenty eight days just after surgery, the left clipped kidney was atrophic, whereas the proper nonclipped kidney displayed compensatory hyper trophy inside the 2K1C mice. Interestingly, sildenafil not simply decreased renal atrophy but additionally original site attenuated the compensa tory hypertrophy. Figure 1 shows the average values of resting MAP and HR measurements in conscious animals 28 days following renal artery clipping. As anticipated, the 2K1C mice showed higher MAP than the Sham mice, as well as the 2K1C mice treated with sildenafil selelck kinase inhibitor showed MAP levels statistically similar to these observed inside the Sham mice. The resting HR of the 2K1C mice was drastically larger than that in ob served in Sham mice and sildenafil remedy abolished this tachycardia. Figure 2 shows typical values of intrarenal angiotensin II in clipped kidneys within the 3 groups of ani mals.
Angiotensin II levels in 2K1C mice were considerably augmented when compared with Sham mice. The 2K1C mice treated with sildenafil exhibited a reduction of 50% in these levels. Renal function The analysis of biochemical parameters showed that soon after 28 days of renovascular hypertension the kidney function was properly preserved. We didn’t observe signifi cant differences in plasma creatinine, urea, pro teinuria and also the esti mated glomerular filtration price involving groups. Sildenafil therapy did not modify those parameters in 2K1C mice. Cell viability assay We evaluated the effect of clipping induced stenosis on renal cell viability utilizing flow cytometry of PI labeled cells. As summarized in Table 1, the stenotic kidneys in the 2K1C group displayed lowered cell viability, which was restored with sildenafil therapy. ROS production ROS production was assessed utilizing flow cytometry with DHE, DCF DA and DAF 2 DA to quantify the produc tion of O2, H2O2 and NO, respectively. Typical histo grams from the flow cytometric analysis showed a rightward shift inside the log of DHE and DCF fluorescence inside the 2K1C group compared together with the Sham group, contrasting using the benefits with the 2K1C sildenafil group.