Experimental study on the effect of anti-microRNA-21 treatment on renal injury in a diabetic nephropathy mouse model
Objective To investigate the effect of anti-microRNA-21(miR-21)treatment on renal injury in a diabetic nephropathy(DN)mouse model.Methods Eight-week-old C57BL/6 mice were randomly divided into the control group,DN group,DN+miR-21 antisense oligonucleotides(ASO)group(DN+miR-21-ASO group),and DN+miR-21 antisense oligonucleotides mismatch base pair(miR-21-mismatched-ASO)group(DN+miR-21-MM-ASO group).DN mouse model was established by intraperitoneal injection of streptozotocin.The 24 h urinary protein(Upro),serum creatinine(Scr),and blood urea nitrogen(BUN)levels in mice were detected using a fully automatic biochemical analyzer.Periodic acid-Schiff(PAS)staining was used to observe glycogen deposition in renal tissues of mice,hematoxylin-eosin(HE)staining was used to observe pathological changes in renal tissues of mice,and TUNEL staining was used to observe apoptosis in renal tissue cells of mice.The relative expression of miR-21 in renal tissue of mice was quantified using real-time quantitative PCR.The protein expression levels of B-cell lym-phoma-2(Bcl-2),Bcl-2 associated X protein(Bax),and cleaved caspase-3 were detected using Western blotting.Results Compared with the control group,the DN group showed significantly upregulated expression of miR-21 in renal tissue[(3.23±0.24)vs.(1.00±0.11),P<0.01],increased 24 h Upro[(53.87±5.87)mg vs.(13.83±1.80)mg,P<0.01],Scr[(89.35±3.34)μmol/L vs.(28.27±2.91)μmol/L,P<0.01],and BUN levels[(25.79±2.15)mmol/L vs.(7.36±0.50)mmol/L,P<0.01].Compared with DN+miR-21-MM-ASO group,the DN+miR-21-ASO group showed inhibited expression of miR-21 in kidney tissue[(0.77±0.11)vs.(3.52±0.27),P<0.01],and significantly decreased 24 h Upro[(29.92±6.06)mg vs.(50.69±6.93)mg,P<0.01],Scr[(68.39±3.75)μmol/L vs.(87.84±6.63)μmol/L,P<0.01]and BUN levels[(17.66±1.04)mmol/L vs.(23.80±3.16)mmol/L,P<0.01].Compared with the control group,PAS staining showed that the DN group mice had increased PAS-positive substances in the mesangial and basement membrane regions of renal tissue,with a darker coloration.Compared with the DN+miR-21-MM-ASO group,the DN+miR-21-ASO group exhibited a significant reduction in PAS-positive substances in the mesangial and basement mem-brane regions of renal tissue,with fewer red-stained areas.Compared with the control group,HE staining showed that the DN group showed renal injury including enlarged glomerular volume and brush border detachment of renal tubules.Compared with the DN+miR-21-MM-ASO group,the DN+miR-21-ASO group showed significant improve-ment in renal injury.In addition,compared with the control group,TUNEL staining showed that the DN group had a significant increase in cell apoptosis in renal tissue cells.Compared with DN+miR-21-MM-ASO group,the DN+miR-21-ASO group showed a significant decrease in renal cell apoptosis.Compared with the control group,West-ern blotting showed that the DN group had upregulated expression of Bax[(3.24±0.26)vs.(1.00±0.10),P<0.01]and cleaved caspase-3[(2.82±0.16)vs.(1.00±0.13),P<0.01],and downregulated expression of Bcl-2[(1.06±0.12)vs.(4.16±0.41),P<0.01].Compared with the DN+miR-21-MM-ASO group,the DN+miR-21-ASO group showed down-regulated expression of Bax[(2.04±0.19)vs.(3.20±0.07),P<0.01]and cleaved caspase-3[(1.54±0.11)vs.(3.10±0.13),P<0.01],and upregulated expression of Bcl-2[(2.05±0.14)vs.(1.02±0.09),P<0.01].Conclusion Anti-miR-21 treatment can significantly alleviate renal injury in diabetic nephropathy mouse model.
万方数据
维普
