Detecting apoptosis signal-regulating kinase 1 protein expression in diabetica nephropathy rats using magnetic hydrophilic nanomaterials

Abstract

Apoptosis signal-regulating kinase 1 (ASK1) signaling is involved in the occurrence and progression of diabetic nephropathy (DN) renal fibrosis, yet the detection of ASK1 requires further investigation. To explore the effect of magnetic nanomaterials combined with ASK1 signaling on renal fibrosis, a DN rat model was generated. Thirty Wistar rats were randomly allocated into three experimental models: control, sham-operated, and DN model groups. Concentrations of postoperative serum creatinine (Scr) and blood urea nitrogen (BUN) were measured to determine the success of the DN model group. Chitosan coated magnetic hydrophilic nanomaterials (Fe3O4@CS MCNCs) were prepared by a “one-pot reaction” method, and further characterized to assess their protein adsorption properties. ASK1 protein expression was analyzed by combining conventional immunohistochemistry and Western blotting techniques with the magnetic nanomaterials. Serum Scr and BUN levels in the DN model group were significantly increased (P < 0.01), indicating that the model was successfully generated. Fe3O4@CS MCNCs were characterized and demonstrated effective hydrophilicity, magnetism, and protein adsorption capacity, highlighting its potential application in proteomic research. The detection of ASK1 protein expression was consistent with that of immunohistochemistry and Western blotting when using the magnetic nanomaterials. Furthermore, ASK1 protein expression in the DN model group was significantly increased compared with the control and sham-operated groups. In this study, magnetic hydrophilic nanomaterials were demonstrated to be feasible for protein detection. In addition, the results indicated the potential involvement of ASK1 signaling in the process of renal fibrosis in DN rats.