title: Isolation and evaluation of the myorelaxant effect of bergapten on isolated rat jejunum.
abstract: BACKGROUND
  Plants of the genus Heracleum L. (Apiaceae) have a long history of being used in traditional medicines for the treatment of alimentary tract disorders, and these biological effects have been ascribed to the presence of furanocoumarins (including bergapten).
  
  
  OBJECTIVE
  This study aimed to develop an efficient, preparative, counter-current chromatographic separation of bergapten in order to characterize its spasmolytic activity in isolated rat jejunum strips.
  
  
  METHODS
  Successful separation of the dichloromethane extract of the fruits of Heracleum leskovii Grossh. was achieved by high-performance countercurrent chromatography (HPCCC) using a two-phase solvent system composed of n-heptane/EtOAc/MeOH/H2O (6:5:6:5, v/v/v/v). The pharmacological assessment of bergapten (0.0001-50 μM) on jejunum smooth muscle strips isolated from rats was conducted under isotonic conditions, following up to three hours of incubation.
  
  
  RESULTS
  The separation method was scaled up six-fold from analytical to semi-preparative conditions, affording bergapten of >99% purity in less than 30 min. This permitted bergapten to be available in quantity for spasmolytic tests on isolated jejunum strips from rats. Bergapten caused myorelaxation of the intestine preparations in the concentration range of 0.0001-1 μM. At higher doses, bergapten caused either relaxation or contraction of the smooth muscle.
  
  
  CONCLUSIONS
  Bergapten was successfully isolated by rapid HPCCC and its spasmolytic activity was confirmed, thereby providing a preliminary evidence base for the traditional medicine application. The data suggest that bergapten causes no irreversible changes to intestinal tissue.
authors.affiliation: a Department of Pharmacognosy with Medicinal Plants Unit , Medical University of Lublin , Lublin , Poland and.
authors.forename: Krystyna
authors.initials: K
authors.lastname: Skalicka-Woźniak
authors.affiliation: b Division of Pharmacology and Toxicology, Faculty of Veterinary Medicine , Warsaw University of Life Sciences , Warsaw , Poland.
authors.forename: Marta
authors.initials: M
authors.lastname: Mendel
authors.affiliation: b Division of Pharmacology and Toxicology, Faculty of Veterinary Medicine , Warsaw University of Life Sciences , Warsaw , Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Chłopecka
authors.affiliation: b Division of Pharmacology and Toxicology, Faculty of Veterinary Medicine , Warsaw University of Life Sciences , Warsaw , Poland.
authors.forename: Natalia
authors.initials: N
authors.lastname: Dziekan
mesh_terms: D000078223:5-Methoxypsoralen; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D005638:Fruit; D005765:Gastrointestinal Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D030002:Heracleum / Q000737:chemistry*; D066298:In Vitro Techniques; D007583:Jejunum / Q000187:drug effects* / Q000502:physiology; D008730:Methoxsalen / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D008752:Methylene Chloride / Q000737:chemistry; D009126:Muscle Relaxation / Q000187:drug effects*; D009130:Muscle, Smooth / Q000187:drug effects* / Q000502:physiology; D010276:Parasympatholytics / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D008517:Phytotherapy; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D010946:Plants, Medicinal; D017208:Rats, Wistar; D012997:Solvents / Q000737:chemistry; D013997:Time Factors
chemical_list: D005765:Gastrointestinal Agents; D010276:Parasympatholytics; D010936:Plant Extracts; D012997:Solvents; D000078223:5-Methoxypsoralen; D008752:Methylene Chloride; D008730:Methoxsalen
keywords: Heracleum leskovii; High-performance counter-current chromatography; intestinal smooth muscle; spasmolytic activity