Renal damage following Alloxan‐induced diabetes is associated with generation of reactive oxygen species, alterations of p53, TGF‐β1, and extracellular matrix metalloproteinases in rats

Abstract

Renal damage is a common problem in diabetes. Alloxan, a potent hyperglycemic and diabetogenic molecule, can induce diabetes through oxidative stress-related mechanisms. Here, we hypothesize that "Alloxan-induced renal damage is associated with alterations of p53, TGF-β1, and extracellular matrix metalloproteinases." To test our hypothesis, we established an animal model (male abino rats) and induced diabetes by intraperitoneal injection of Alloxan monohydrate. Rats with fasting blood glucose level ≥ 200 mg/dL were considered diabetic and were sacrificed after 14, 28, and 42 day intervals. Tissue levels of malondialdehyde and glutathione levels (markers of oxidative stress), and serum MMP-1 were measured. The expression patterns of p53, TGF-β1were evaluated using Western blot and immunohistochemical methods. TIMP-1 expression pattern was determined using RT-PCR and immunohistochemical methods. Alloxan treatment induced histological features of renal damage (inflammation and fibrosis) and was associated with deterioration of the renal functions (elevated blood urea nitrogen and creatinin levels), hyperglycemia, and oxidative stresss (increased malondialdehyde and decreased glutathione levels). There was over-expression of the TGF-β1 protein (profibrogenic protein), p53 (proapoptotic protein), and alterations of extracellular matrix proteins (low level of serum MMP-1 and over-expression of TIMP-1). Alterations of TGF-β, p53, and extracellular matrix metalloproteinases contribute to the pathogenesis of Alloxan-induced renal damage.

Transforming Growth Factor-Beta1 in Diabetic Kidney Disease

“…The abundance of matrix metalloproteinase-9 (MMP-9), an ECM-degradation MMP, was decreased in transgenic mice that overexpressed TGF-β1 (Zechel et al, 2002;Ueberham et al, 2003). In addition, TGF-β1 augmented the expression of tissue inhibitor of metalloproteinases-1 (Ueberham et al, 2003;Abdel Aziz et al, 2017), which inhibited the ECM-degrading MMPs.…”

“…Although sodium-glucose co-transporter 2 (SGLT2) inhibitors have conferred cardiovascular and renal protection (Perkovic et al, 2019), effective therapy for DKD is still unavailable. An epidemiological study revealed that the 5-year mortality rate of DKD was approximately 40%, as high as many cancers (Abdel Aziz et al, 2017). Transforming growth factor-β1 (TGF-β1) signaling contributes to DKD progression, and inhibiting TGF-β1 signaling has shown potential renoprotective properties in animal and human studies.…”

Matrix Metalloproteinases in Diabetic Kidney Disease

“…Unlikely to enalapril [74], insulin treatment ameliorated these changes [75]. MMP-1 expression and serum levels were also down-regulated in alloxan induced-diabetic mice [76,77], while local delivery of Mmp-1 into mice kidneys prevented diabetic renal fibrosis [78].…”

Matrix Metalloproteinases in Renal Diseases: A Critical Appraisal

“…Another experimental study with alloxan-induced diabetes in rodents was associated with low serum MMP-1 levels and increased levels of TIMP-1 expression in promoting kidney damage [31]. …”