【Animal Modeling】-miR-122 Expression Changes and Action Mechanisms in Kidney Tissues of Mice with Diabetic Nephropathy

  Objective To investigate the expression changes of miR-122 in the kidney tissue of diabetic nephropathy mice and the effect and possible mechanism of silencing miR-122 on the kidney tissue of DN mice.

  Methods The DN model of C57BL/6 mice was established by intraperitoneal injection of streptozotocin (STZ) combined with a high-sugar and high-fat diet. Thirty DN mice with successful modeling were randomly divided into model group, antagomir-NC group, and antagomir-122 group, with 10 mice in each group, and 10 untreated healthy C57BL/6 mice were selected as the normal control group. . After successful modeling, antagomir-122 group and antagomir-NC group were given tail vein injection of antagomir-122 and antagomir-NC every 7 days for intervention, and model group and normal control group were given tail vein injection of equal volume of normal saline instead. After 8 weeks, the serum and urine were collected, and the mice were sacrificed to obtain kidney specimens. Serum creatinine (Cr), blood urea nitrogen (BUN) and 24-hour urine protein quantitative levels were detected by automatic biochemical analyzer, and renal tissue was evaluated by periodic acid Schiff (PAS) staining. Pathological changes, qRT-PCR was used to detect the expression level of miR-122 in renal tissue, and kits were used to detect glutathione peroxidase (GSH-Px), malondialdehyde (MDA), superoxide dismutase (SOD) in renal tissue ) level, Western blot was used to detect the expression levels of Sirt1, α-SMA and fibronectin proteins in kidney tissue.

  Results Compared with the normal control group, the glomerular sclerosis score of the mice in the model group increased (P<0.05), and the expressions of miR-122, α-SMA and fibronectin protein in the kidney tissue were significantly increased (P<0.05). Serum Cr and BUN levels, 24-hour urine protein quantitative levels, and renal tissue MDA levels were significantly increased (P<0.05), while renal tissue GSH-Px and SOD levels and Sirt1 protein expression levels were significantly decreased (P<0.05) .="" compared="" with="" there="" was="" no="" significant="" difference="" in="" each="" index="" between="" the="" model="" group="" and="" antagomir-nc="" p="">0.05). Compared with the antagomir-NC group, the glomerular sclerosis score of the mice in the antagomir-122 group decreased (P<0.05), and the expressions of miR-122 and α-SMA and fibronectin proteins in the kidney tissue were significantly decreased (P<0.05). , serum Cr, BUN levels, 24-h urine protein quantitative level, renal tissue MDA levels were significantly decreased (P<0.05), renal tissue GSH-Px and SOD levels, Sirt1 protein expression levels were significantly increased (P<0.05). 05).

  Conclusion The expression of miR-122 is increased in the kidney tissue of DN mice. Silencing miR-122 has obvious protective effect on kidney tissue of DN mice through anti-oxidative stress and fibrosis, and the mechanism may be related to the regulation of Sirt1 gene.