The anti-inflammation effect of Moutan Cortex on advanced glycation end products-induced rat mesangial cells dysfunction and High-glucose-fat diet and streptozotocin-induced diabetic nephropathy rats

Abstract

Ethnopharmacological relevance: Moutan Cortex (MC, family: Paeonia suffruticosa Andr.) is a well-known traditional herbal medicine that has been shown to hold a protective effect on inflammation in several diseases. However, its anti-inflammatory activity on diabetic nephropathy (DN) has been less reported. The present study was conducted to evaluate the potential attenuation activities of MC on inflammation in AGEs-induced rat mesangial cells dysfunction and high-glucose-fat diet and streptozotocin (STZ)-induced DN rats and explore the possible mechanism underlying its DN effect.

Materials and methods: The inflammation in mesangial cells (HBZY-1) was induced by 200 μg/ml advanced glycation end products (AGEs). DN rats model was established by an administration high-glucose-fat diet and an intraperitoneal injection of STZ (30 mg/kg). Interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) level in cell supernatant and rats serum were detected by appropriate kits. A co-culture system of mesangial cells and macrophages was performed to evaluate the migration of macrophages. Immunohistochemical assay was applied to examine transforming growth factor beta1 (TGF-β1), IL-6, MCP-1 and intercellular adhesion molecule-1 (ICAM-1) expression in kidney tissues of rats. Furthermore, western blot analysis was carried out to examine TGF-β1, IL-6, MCP-1, ICAM-1 and RAGE protein expressions in mesangial cells.

Results: Pretreatment with MC could significantly inhibit AGEs-induced migration of macrophages in the co-culture system of mesangial cell and macrophage. MC could decrease IL-6 and MCP-1 levels in serum of DN rats in a dose-dependent manner. Furthermore, MC also improved the blood glucose, serum creatinine and urine protein levels. Both immunocytochemistry analysis and western blot analysis showed that MC decreased significantly the over-expression of IL-6, MCP-1, TGF-β1, ICAM-1 and RAGE in mesangial cells or kidney tissues. Additionally, the protein expression of proinflammatory cytokine could also be down-regulated by the pretreatment of RAGE-Ab (5 μg/ml).

Conclusion: These findings indicated that the extract of MC had an amelioration activity on the inflammation in AGEs-induced mesangial cells dysfunction and high-glucose-fat diet and STZ-induced DN rats. The protective effect might be associated with the intervention of MC via target of RAGE. These findings suggested that MC might be a benefit agent for the prevention and treatment of DN.

Keywords: AG; AGEs; BSA; DMEM; DN; Diabetic nephropathy; Dulbecco's modified Eagle's medium; FBS; ICAM-1; IL-6; Inflammation; MC; MCP-1; Moutan Cortex; Moutan cortex; OD; OS; PBS; RAGE; ROS; Rage; SD; STZ; TGF-β1; advanced glycation end products; aminoguanidine; bovine serum albumin; diabetic nephropathy; fetal bovine serum; intercellular adhesion molecule-1; interleukin-6; monocyte chemoattractant protein-1; optical density; oxidative stress; phosphate buffered saline; reactive oxygen species; receptor for AGEs; standard deviation; streptozotocin; transforming growth factor beta1.