title: 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: BACKGROUND
  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.
  
  
  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).
  
  
  CONCLUSIONS
  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.
authors.affiliation: Key Laboratory of New Drug Delivery Systems of Chinese Meteria Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing 210028, PR China; Department of Pharmaceutics, Jiangsu University, Jiangsu, Zhenjiang 212013, PR China.
authors.forename: Ming-hua
authors.initials: MH
authors.lastname: Zhang
authors.affiliation: Key Laboratory of New Drug Delivery Systems of Chinese Meteria Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing 210028, PR China. Electronic address: wenmoxiushi@163.com.
authors.forename: Liang
authors.initials: L
authors.lastname: Feng
authors.affiliation: Key Laboratory of New Drug Delivery Systems of Chinese Meteria Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing 210028, PR China; Nanjing Institute of Supervision & Testing on Product Quality, Jiangsu, Nanjing 210028, PR China.
authors.forename: Mao-mao
authors.initials: MM
authors.lastname: Zhu
authors.affiliation: Key Laboratory of New Drug Delivery Systems of Chinese Meteria Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing 210028, PR China; College of Pharmacy, Nanjing University of Chinese Medicine, Jiangsu, Nanjing 210046, PR China.
authors.forename: Jun-fei
authors.initials: JF
authors.lastname: Gu
authors.affiliation: Key Laboratory of New Drug Delivery Systems of Chinese Meteria Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing 210028, PR China; Nanjing Institute of Supervision & Testing on Product Quality, Jiangsu, Nanjing 210028, PR China; College of Pharmacy, Nanjing University of Chinese Medicine, Jiangsu, Nanjing 210046, PR China.
authors.forename: Jun
authors.initials: J
authors.lastname: Jiang
authors.affiliation: Key Laboratory of New Drug Delivery Systems of Chinese Meteria Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing 210028, PR China; College of Pharmacy, Nanjing University of Chinese Medicine, Jiangsu, Nanjing 210046, PR China.
authors.forename: Xu-dong
authors.initials: XD
authors.lastname: Cheng
authors.affiliation: Key Laboratory of New Drug Delivery Systems of Chinese Meteria Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing 210028, PR China; College of Pharmacy, Nanjing University of Chinese Medicine, Jiangsu, Nanjing 210046, PR China.
authors.forename: Shu-ming
authors.initials: SM
authors.lastname: Ding
authors.affiliation: Key Laboratory of New Drug Delivery Systems of Chinese Meteria Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing 210028, PR China; College of Pharmacy, Nanjing University of Chinese Medicine, Jiangsu, Nanjing 210046, PR China.
authors.forename: Chan
authors.initials: C
authors.lastname: Wu
authors.affiliation: Key Laboratory of New Drug Delivery Systems of Chinese Meteria Medica, Jiangsu Provincial Academy of Chinese Medicine, Jiangsu, Nanjing 210028, PR China; Department of Pharmaceutics, Jiangsu University, Jiangsu, Zhenjiang 212013, PR China; College of Pharmacy, Nanjing University of Chinese Medicine, Jiangsu, Nanjing 210046, PR China. Electronic address: jxiaobin2005@hotmail.com.
authors.forename: Xiao-bin
authors.initials: XB
authors.lastname: Jia
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000494:pharmacology; D002460:Cell Line; D003921:Diabetes Mellitus, Experimental / Q000150:complications*; D003928:Diabetic Nephropathies / Q000188:drug therapy; D004041:Dietary Fats / Q000009:adverse effects*; D004305:Dose-Response Relationship, Drug; D004365:Drugs, Chinese Herbal / Q000737:chemistry / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D005947:Glucose / Q000008:administration & dosage / Q000009:adverse effects*; D017127:Glycation End Products, Advanced / Q000633:toxicity*; D050527:Mesangial Cells / Q000187:drug effects* / Q000378:metabolism; D029596:Paeonia / Q000737:chemistry; D011897:Random Allocation; D051381:Rats
chemical_list: D000893:Anti-Inflammatory Agents; D004041:Dietary Fats; D004365:Drugs, Chinese Herbal; D017127:Glycation End Products, Advanced; C038881:moutan cortex; D005947:Glucose
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