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Int J Biol Sci 2014; 10(2):149-159. doi:10.7150/ijbs.7875

Research Paper

Establishment of an Inflamed Animal Model of Diabetic Nephropathy

Kun Ling Ma1,✉, Yang Zhang1, Jing Liu1, Yu Wu1, Ze Bo Hu1, Xiong Zhong Ruan2, Bi Cheng Liu1

1. Institute of Nephrology, Zhong Da Hospital, Southeast University School of Medicine, Nanjing City, Jiangsu Province, China.
2. Centre for Nephrology, University College London (UCL) Medical School, Royal Free Campus, UK.

Abstract

Aims Inflammatory stress plays a crucial role in the progression of diabetic nephropathy (DN). This study aimed to establish a novel inflamed animal model of DN and to evaluate its significance in DN.

Methods Nondiabetic db/m mice and diabetic db/db mice were randomly divided into four groups: db/m, db/m+casein, db/db, and db/db+casein for eight weeks. Casein was subcutaneously injected to induce chronic inflammation. Body weight and albumin to creatinine ratio (ACR) in the urine were measured every week. The plasma levels of serum amyloid protein A (SAA) and tumour necrotic factor-α (TNF-α) were determined with the enzyme-linked immunosorbent assay. The morphological changes to the renal pathology and ultra-microstructures were checked by pathological staining and electron microscopy. Immunofluorescent staining and Western blotting were used to determine the protein expression of podocyte-specific molecules and inflammatory cytokines in kidneys.

Results ACR, plasma levels of SAA and TNF-α, protein expression of inflammatory cytokines, mesangial expansion, collagen accumulation, and foot process effacement in kidneys of casein-injected db/db mice were significantly increased compared with the db/db mice. Casein injection markedly decreased the protein expression of Wilms' tumor-1 and nephrin in kidneys of db/db mice, which are specific podocyte biomarkers, suggesting that chronic inflammation accelerates podocyte injuries in db/db mice. Interestingly, no obvious urinary protein, inflammatory cytokine expression, or histological changes in the kidneys of casein-injected db/m mice were found compared with the db/m mice.

Conclusion An inflamed animal model of DN was successfully established and may provide a useful tool for investigating the pathogenesis of DN under inflammatory stress.

Keywords: Diabetic nephropathy, inflammation, db/db mice, animal model

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How to cite this article:
Ma KL, Zhang Y, Liu J, Wu Y, Hu ZB, Ruan XZ, Liu BC. Establishment of an Inflamed Animal Model of Diabetic Nephropathy. Int J Biol Sci 2014; 10(2):149-159. doi:10.7150/ijbs.7875. Available from http://www.ijbs.com/v10p0149.htm