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Organelle movement in melanophores: Effects of Panax ginseng, ginsenosides and quercetin
Linköping University, Department of Medical and Health Sciences, Pharmacology. Linköping University, Faculty of Health Sciences.
2009 (English)Licentiate thesis, comprehensive summary (Other academic)
Abstract [en]

Panax ginseng is a traditional herb that has been used for over 2000 years to promote health and longevity. Active components of ginseng include ginsenosides, polysaccharides, flavonoids, polyacetylenes, peptides, vitamins, phenols and enzymes, of which the ginsenosides are considered to be the major bioactive constituents. Although widely used, the exact mechanisms of ginseng and its compounds remain unclear. In this thesis we use melanophores from Xenopus laevis to investigate the effects of Panax ginseng extract G115 and its constituents on organelle transport and signalling. Due to coordinated bidirectional movement of their pigmented granules (melanosomes), in response to defined chemical signals, melanophores are capable of fast colour changes and provide a great model for the study of intracellular transport. The movement is regulated by alterations in cyclic adenosine 3’:5’-monophosphate (cAMP) concentration, where a high or low level induce anterograde (dispersion) or retrograde (aggregation) transport respectively, resulting in a dark or light cell. Here we demonstrate that Panax ginseng and its constituents ginsenoside Rc and Rd and flavonoid quercetin induce a concentration-dependent anterograde transport of melanosomes. The effect of ginseng is shown to be independent of cAMP changes and protein kinase A activation. Upon incubation of melanophores with a combination of Rc or Rd and quercetin, a synergistic increase in anterograde movement was seen, indicating cooperation between the ginsenoside and flavonoid parts of ginseng. Protein kinase C (PKC) inhibitor Myristoylated EGF-R Fragment 651-658 decreased the anterograde movement stimulated by ginseng and ginsenoside Rc and Rd. Moreover, ginseng, but not ginsenosides or quercetin, stimulated an activation of 44/42-mitogen activated protein kinase (MAPK), previously shown to be involved in both aggregation and dispersion of melanosomes. PKC-inhibition did not affect the MAPK-activation, suggesting a role for PKC in the ginseng- and ginsenoside-induced dispersion but not as an upstream activator of MAPK.

Abstract [sv]

Panax ginseng är ett av de vanligaste naturläkemedlen i världen och används traditionellt för att öka kroppens uthållighet, motståndskraft och styrka. Ginseng är ett komplext ämne bestående av ett antal olika substanser, inklusive ginsenosider, flavonoider, vitaminer och enzymer, av vilka de steroidlika ginsenosiderna anses vara de mest aktiva beståndsdelarna. Flavonoider (som finns i till exempel frukt och grönsaker) och ginseng har genom forskning visat sig motverka bland annat hjärt-och kärlsjukdomar, diabetes, cancer och demens. Trots den omfattande användningen är dock mekanismen för hur ginseng verkar fortfarande oklar. I den här studien har vi använt pigmentinnehållande celler, melanoforer, från afrikansk klogroda för att undersöka effekterna av Panax ginseng på pigment-transport och dess maskineri. Melanoforer har förmågan att snabbt ändra färg genom samordnad förflyttning av pigmentkorn fram och tillbaka i cellen, och utgör en utmärkt modell för studier av intracellulär transport. Förflyttningen regleras av förändringar i halten av cykliskt adenosin-monofosfat (cAMP) i cellen, där en hög eller låg koncentration medför spridning av pigment över hela cellen (dispergering) eller en ansamling i mitten (aggregering), vilket resulterar i mörka respektive ljusa celler. Här visar vi att Panax ginseng, ginsenosiderna Rc och Rd samt flavonoiden quercetin stimulerar en dispergering av pigmentkornen. När melanoforerna inkuberades med en kombination av ginsenosid Rc eller Rd och quercetin, kunde en synergistisk ökning av dispergeringen ses, vilket tyder på en samverkan mellan ginsenosid- och flavonoid-delarna av ginseng. Ett protein som tidigare visats vara viktigt för pigmenttransporten är mitogen-aktiverat protein kinas (MAPK), och här visar vi att också melanoforer stimulerade med ginseng, men dock inte med ginsenosider eller quercetin, innehåller aktiverat MAPK. Genom att blockera enzymet protein kinas C (PKC) (känd aktivator av dispergering), minskade den ginseng- och ginsenosid-inducerade dispergeringen, medan aktiveringen av MAPK inte påverkades alls. Detta pekar på en roll för PKC i pigment-transporten men inte som en aktivator av MAPK.

Place, publisher, year, edition, pages
Linköping: Linköping University Electronic Press , 2009. , p. 61
Series
Linköping Studies in Health Sciences. Thesis, ISSN 1100-6013 ; 103
Keywords [en]
melanophores, Panax ginseng, ginsenosides, quercetin, organell anterograde transport, endothelin-3, mitogen activated protein kinase, protein kinase C
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:liu:diva-19973ISBN: 978-91-7393-581-4 (print)OAI: oai:DiVA.org:liu-19973DiVA, id: diva2:232380
Presentation
2009-09-10, Eken, Campus US, Linköpings universitet, Linköping, 13:00 (Swedish)
Opponent
Supervisors
Available from: 2009-09-22 Created: 2009-08-21 Last updated: 2020-03-10Bibliographically approved
List of papers
1. Panax ginseng induces anterograde transport of pigment organelles in Xenopus melanophores
Open this publication in new window or tab >>Panax ginseng induces anterograde transport of pigment organelles in Xenopus melanophores
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2008 (English)In: Journal of Ethnopharmacology, ISSN 0378-8741, E-ISSN 1872-7573, Vol. 119, no 1, p. 17-23Article in journal (Refereed) Published
Abstract [en]

Melanophores from Xenopus laevis are pigmented cells, capable of quick colour changes through cyclic adenosine 3':5'-monophosphate (cAMP) coordinated transport of their intracellular pigment granules, melanosomes. In this study we use the melanophore cell line to evaluate the effects of Panax ginseng extract G115 on organelle transport. Absorbance readings of melanophore-coated microplates, Correlate-EIA direct cAMP enzyme immunoassay kit, and western blot were used to measure the melanosome movement and changes in intracellular signalling. We show that Panax ginseng induces a fast concentration-dependent anterograde transport of the melanosomes. No significant increase in the cAMP level was seen and pre-incubation of melanophores with the protein kinase C (PKC) inhibitor EGF-R Fragment 651-658 (M-EGF) only partly decreased the ginseng-induced dispersion. We also demonstrate that Panax ginseng, endothelin-3 (ET-3) and alpha-melanocyte stimulating hormone (MSH) stimulate an activation of mitogen activated protein kinase (MAPK). Pre-incubation with M-EGF decreased the MAPK activity induced by ET-3 and MSH, but again only marginally affected the response of Panax ginseng. Thus, in melanophores we suggest that Panax ginseng stimulates an anterograde transport of pigment organelles via a non-cAMP and mainly PKC-independent pathway.

Place, publisher, year, edition, pages
Elsevier, 2008
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-19968 (URN)10.1016/j.jep.2008.05.024 (DOI)
Available from: 2009-08-21 Created: 2009-08-21 Last updated: 2017-12-13Bibliographically approved
2. Role of ginsenosides and quercetin in anterograde transport of melanosomes
Open this publication in new window or tab >>Role of ginsenosides and quercetin in anterograde transport of melanosomes
(English)Manuscript (preprint) (Other academic)
Abstract [en]

Panax ginseng is a traditional herb that has been used in the Orient for several thousands of years as a tonic and restorative. Active components of ginseng include ginsenosides, polysaccharides, flavonoids, polyacetylenes, peptides, vitamins, phenols and enzymes, of which the ginsenosides are considered to be the major bioactive constituents. In spite of extensive use, the exact mechanisms of ginseng and its components are still unknown. In the present study we use melanophores from Xenopus laevis to investigate the effects of Panax ginseng extract G115, ginsenosides and the flavonoid quercetin on pigment organelle transport and signalling. Through coordinated transport of their black pigmented granules (melanosomes), melanophores are capable of fast colour changes. The movement is regulated by changes in cyclic adenosine 3’:5’-monophosphate (cAMP) concentration, where a high or low level induce anterograde (dispersion) or retrograde (aggregation) transport respectively, leaving a dark or light cell. Previously we have shown that Panax ginseng induces dispersion of melanosomes. Here, ginsenoside Rc and Rd and the flavonoid quercetin are shown to stimulate an anterograde transport of pigment organelles. When Rc or Rd and quercetin were combined, a significant synergistic increase in dispersion could be seen. Protein kinase C (PKC) inhibitor Myristoylated EGF-R Fragment (651-658) (M-EGF) decreased the anterograde movement stimulated by ginseng and ginsenoside Rc and Rd. We also demonstrate that ginseng, but not ginsenosides or quercetin, induce a concentration-dependent activation of 44/42-mitogen activated protein kinase (MAPK). PKC-inhibition did not affect the MAPK-activation, suggesting a role for PKC in the ginseng- and ginsenoside-induced anterograde transport but not in the activation of MAPK.

Keywords
melanophore, pigment organelle transport, Panax ginseng, ginsenosides, quercetin, PKC, MAPK
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-19970 (URN)
Available from: 2009-08-21 Created: 2009-08-21 Last updated: 2010-01-14Bibliographically approved

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