Clopidogrel delays and can reverse diabetic nephropathy pathogenesis in type 2 diabetic db/db mice

Abstract

BACKGROUND Diabetic nephropathy (DN) is the principal cause of end-stage renal disease. Previous studies have shown that clopidogrel can prevent the early progression of renal injury. AIM To elucidate whether clopidogrel is beneficial against DN by using a db / db mouse model. METHODS db/db mice with a higher urinary albumin/creatinine ratio (ACR) relative to age- and sex-matched wild-type control mice were randomly allocated to clopidogrel and vehicle treatment groups. Clopidogrel was administered at doses of 5, 10, and 20 mg/kg by gavage for 12 wk. Body mass, blood glucose level, and urinary creatinine and albumin concentrations in each group were measured before and after the intervention. Renal fibrosis was evaluated using periodic acid-Schiff and Masson’s trichrome staining. The renal protein expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and F4/80 was assessed using immunohistochemistry. Urinary TNF-α, monocyte chemoattractant protein-1 (MCP-1), and IL-6 levels were analyzed using enzyme-linked immunosorbent assay; TNF-α and IL-1β mRNA expression was measured using real-time quantitative polymerase chain reaction. The protein expression of fibronectin (FN) and collagen I was assessed using immunohistochemistry. RESULTS Clopidogrel treatment did not affect the body mass or blood glucose level of the db/db mice; however, it increased bleeding time and reduced urinary ACR in a dose-dependent manner. Immunohistochemical staining revealed an amelioration of renal fibrosis, significantly lower deposition of FN and collagen I, and significantly lower expression of the proinflammatory cytokines TNF-α and IL-1β and lower levels of urinary TNF-α and MCP-1 in the clopidogrel-treated db/db mice ( P < 0.05). Furthermore, clopidogrel significantly reduced macrophage infiltration into the glomeruli of the db/db mice. CONCLUSION Clopidogrel significantly reduced renal collagen deposition and fibrosis and prevented renal dysfunction in db / db mice, most likely through inhibition of renal macrophage infiltration and the associated inflammation.

Mass cytometry reveals the corneal immune cell changes at single cell level in diabetic mice

“…As a well-known type 2 diabetes models, this DB mouse line has a series of characteristics such as significant increases in body weight beginning at 4 weeks, hyperglycemia at 8 weeks, insulinemia (> 3-fold) at 8 weeks ( 15 ). Moreover, the DB corneas showed significant histopathological alterations, with impaired proliferation, decreased innervation at 20-24 weeks ( 16 ), lower density of the corneal subbasal nerve plexus and corneal epithelial branches at 13 weeks ( 17 ).…”

The mechanisms and therapeutic potential of clopidogrel in mitigating diabetic cardiomyopathy in db/db mice

“… 16 Other studies reported that Clo treatment reversed diabetic nephropathy by inhibiting fibrosis and inflammation. 19 , 20 …”