【Animal Modeling】-The effect of aloin on NOX4/ROS/p38 MAPK signaling pathway and podocyte function in rats with diabetic nephropathy

  Objective: To investigate the function of podocytes in diabetic nephropathy (DN) rats and the signal transduction pathway of nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4)/reactive oxygen species (ROS)/p38 mitogen-activated protein kinase (p38MAPK).

  Method: The rats were fed a high-sugar and high-fat diet for 4 weeks, and 40 mg/kg STZ was injected intraperitoneally to establish a DN rat model, model group, and positive control group (low, medium, high). The normal group was not treated with rats. The positive control group was given 9.45 mg/(kg?D) Tangspeil in the stomach, while the (low, medium, and high) doses of the experimental group were given 10, 20 and 40 mg/(kg?D) aloin in the abdominal cavity. Both the normal group and the model group took aloin and administered the same amount of distilled water to the stomach for 6 weeks. Blood glucose meter detects fasting blood glucose level; enzyme-linked immunosorbent assay (ELISA) detects serum interleukin 1β (IL-1β) and tumor necrosis factor-α (TNF-α) HE staining to observe rat kidney tissue morphology; superoxide Dismutase (SOD), malondialdehyde (MDA) and reactive oxygen species (ROS) kits can detect SOD in kidney tissue, MDA and ROS levels Western blotting can detect kidney NOX4, p38MAPK, phosphorylated p38MAPK and renal small The transmembrane proteins nephrin and podosine at the level of the ball and foot.

  Results: Before administration, the fasting blood glucose levels of the model group, the positive control group and the (low, medium, and high) dose experimental group were higher than the normal group (P\u003c0.05). After the administration, the renal glomeruli in the model group were thickened, the glomerular mesangium thickened and the glomerular basement membrane thickened, and the interstitial inflammatory infiltration; (medium and low dose) the experimental group glomerular hypertrophy gradually reduced, The glomerular mesangial hyperplasia was mild to moderate, and the renal tubule dilatation slowed down; the renal tissue in the dose experimental group was normal, with clear structure, and regular glomeruli and tubules. The fasting blood glucose, serum IL-1β and TNF-α, MDA, ROS, NOX4 and phosphorylated p38MAPK protein levels of the model group were higher than those of the normal group (P\u003c0.05), and the SOD level and SOD were increased in the normal group. Kidney tissue/MDA, renin and podosine protein levels decreased (P\u003c0.05). Positive control group, high-dose experimental group, fasting blood glucose, serum IL-1β, TNF-α, kidney tissue MDA, ROS, NOX4, phosphorylated p38MAPK protein levels were lower than the model group (P\u003c0).05), middle-dose experiment The protein levels of SOD, SOD/MDA, nephrin and Podosine in the kidney tissue of the group increased (P\u003c0.05); fasting blood glucose and serum IL-1β decreased. MDA, ROS and NOX4 protein in kidney tissue (P\u003c0.05), P\u003c0.05), while the level of SOD, SOD/MDA, renin and Podosine protein in kidney tissue increased (P\u003c0.05) ). The low-dose experimental group had fasting blood glucose. , Serum IL-1β, TNF-α, renal tissue MDA, NOX4 protein levels decreased (P\u003c0.05); renal tissue SOD levels increased (P\u003c0.05). Compared with before administration, the fasting blood glucose levels of the positive control group and the (low, medium, and high) dose experimental groups were reduced.

  Conclusion: Aloin may down-regulate the NOX4/ROS/p38 signaling pathway, thereby achieving anti-inflammatory and podocyte recovery.