title: Biomolecular evidence of anti-inflammatory effects by Clematis mandshurica Ruprecht root extract in rodent cells.
abstract: BACKGROUND
  Clematis mandshurica Ruprecht root is widely used in Asia as an analgesic and anti-inflammatory agent. This research investigated the anti-inflammatory effects of Clematis mandshurica Ruprecht root extract (CRE) using RAW 264.7 macrophage cells and carrageenan- (CA-) induced rat paw edema.
  
  
  METHODS
  Production of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, nitric oxide (NO) and prostaglandin E2 (PGE2) in the culture supernatant, mRNA expression of TNF-α, IL-1β, IL-6, iNOS and COX-2, protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs) in the extract were assayed. In addition, the effect of CRE on acute inflammation in vivo was observed using CA-induced rat hind paw edema assay. The changes on the histopathology and histomorphometry of hind paw skins-dorsum and ventrum pedis were observed using CA-treated rats.
  
  
  RESULTS
  Treatment with CRE (0.25, 0.5, and 1 mg/mL) resulted in inhibited levels of protein expression of lipopolysaccharide- (LPS-) induced iNOS, COX-2, NF-κB, and MAPKs (ERK, JNK, and p38) as well as production of TNF-α, IL-1β, IL-6, NO, and PGE2 induced by LPS. Consistent with these results, CRE reduced the LPS-induced expressions of these cytokines, iNOS and COX-2 at the mRNA levels in a dose-dependent manner. In particular, results of the CA-induced rat hind paw edema assay showed an anti-edema effect of CRE. In addition, treatment with CRE resulted in dose-dependent inhibition of CA-induced increases of skin thickness, mast cell degranulation, and infiltrated inflammatory, TNF-α, IL-1β, iNOS, and COX-2-positive cells in both dorsum and ventrum pedis skin, respectively.
  
  
  CONCLUSIONS
  These results demonstrate that CRE exhibits anti-inflammatory activities via decreasing production of pro-inflammatory mediators through suppression of the pathways of NF-κB and MAPKs in LPS-induced macrophage cells. In addition, results of the CA-induced rat hind paw edema assay show an anti-edema effect of CRE. Our findings also support the traditional use of CRE in the inflammatory symptoms of rheumatic arthritis and acute icteric hepatitis. Thus, CRE may have therapeutic potential for a variety of inflammation-mediated diseases and may be developed into potent anti-inflammatory drugs.
authors: Institute of Marine BioTechnology, Pusan National University, Busan 609-735, Republic of Korea.
authors: Chul Won
authors: CW
authors: Lee
authors: College of Oriental Medicine, Daegu Haany University, Gyeongsan 712-715, Republic of Korea.
authors: Sang Mi
authors: SM
authors: Park
authors: School of Medicine, Pusan National University, Yangsan 626-870, Republic of Korea.
authors: Youn Sook
authors: YS
authors: Kim
authors: College of Oriental Medicine, Daegu Haany University, Gyeongsan 712-715, Republic of Korea.
authors: Kyung Hwan
authors: KH
authors: Jegal
authors: Department of Pharmaceutical Engineering, Daegu Haany University, Gyeongsan 712-715, Republic of Korea.
authors: Jong Rok
authors: JR
authors: Lee
authors: College of Oriental Medicine, Daegu Haany University, Gyeongsan 712-715, Republic of Korea.
authors: Il Je
authors: IJ
authors: Cho
authors: College of Oriental Medicine, Daegu Haany University, Gyeongsan 712-715, Republic of Korea.
authors: Sae Kwang
authors: SK
authors: Ku
authors: Institute of Marine BioTechnology, Pusan National University, Busan 609-735, Republic of Korea.
authors: Ji Yeon
authors: JY
authors: Lee
authors: Institute of Marine BioTechnology, Pusan National University, Busan 609-735, Republic of Korea.
authors: Yong-Tae
authors: YT
authors: Ahn
authors: Institute of Marine BioTechnology, Pusan National University, Busan 609-735, Republic of Korea.
authors: Yonghae
authors: Y
authors: Son
authors: Biomedical Research Center, Ulsan University Hospital, College of Medicine, University of Ulsan, Ulsan 680-749, Republic of Korea.
authors: Seong A
authors: SA
authors: Ju
authors: College of Oriental Medicine, Daegu Haany University, Gyeongsan 712-715, Republic of Korea. Electronic address: sckim@dhu.ac.kr.
authors: Sang Chan
authors: SC
authors: Kim
authors: Institute of Marine BioTechnology, Pusan National University, Busan 609-735, Republic of Korea; Division of Pharmacology, School of Korean Medicine, Pusan National University, Yangsan 626-870, Republic of Korea. Electronic address: wgan@pusan.ac.kr.
authors: Won G
authors: WG
authors: An
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D002351:Carrageenan / Q000633:toxicity; D002460:Cell Line; D039324:Clematis / Q000737:chemistry*; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004487:Edema / Q000188:drug therapy* / Q000473:pathology; D007249:Inflammation / Q000188:drug therapy / Q000473:pathology; D018836:Inflammation Mediators / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000473:pathology; D008297:Male; D010936:Plant Extracts / Q000008:administration & dosage / Q000494:pharmacology*; D018517:Plant Roots; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D000893:Anti-Inflammatory Agents; D016207:Cytokines; D018836:Inflammation Mediators; D010936:Plant Extracts; D002351:Carrageenan
keywords: Anti-inflammation; Clematis mandshurica Ruprecht; Histological examination; Mitogen-activated protein kinases; Nuclear factor-kappa B; Rat paw edema