Keratinocyte lipid fluidity under the influence of cholesterols, hydrocortisones, "active lipid", tocopherol and retinoic acid--a fluorescence polarization study with regard to physiological and pathophysiological epidermopoiesis and its therapeutic accessibility.
Lipid fluidity of freshly isolated human (H) and guinea pig (GP) keratinocytes (K) was determined as the reciprocal of diphenylhexatriene (DPH) fluorescence polarization (P-value), the temperature being kept at 25 degrees C and cell density standardized to 550,000 per ml (level of statistical significance a less than 0.05). An experimental model involving short-term incubations (2.5 hours, 37 degrees C) of GPK in 1% ethanolic lipid solutions (15 mg lipid agent per ml ethanol) was set up to investigate accumulation a) of cholesterol due to terminal differentiation of keratinocytes and b) of cholesteryl sulfate due to the lack of steroid sulfatase activity in recessive X-linked ichthyosis (RXLI). In comparison to the control including 1% ethanol (P = 0.291 +/- 0.004), significant rigidifying effects were demonstrated for cholesteryl hemisuccinate (0.331 +/- 0.005) and cholesteryl sulfate (0.310 +/- 0.002). Correspondingly, a significant increase of the P-value was also induced by cholesteryl hemisuccinate in HK. Rigidification of GPK by a preincubation with cholesteryl sulfate (P = 0.306 +/- 0.002) could be antagonized by a subsequent short-term incubation with "active lipid (mixture 721)" (0.285 +/- 0.003, a less than 0.05) which may be relevant for future therapeutic strategies in RXLI. Other steran molecules such as hydrocortisone-21-hemisuccinate or hydrocortisone acetate did not affect lipid fluidity. With regard to the therapeutic potency of retinoids in epidermopoietic disorders, incubations of HK with all-trans-retinoic-acid were compared to those with also lipophilic vitamin E, i.e. d-alpha-tocopherol, for 2.5 hours at 37 degrees C using 1% DMSO as a solvent.(ABSTRACT TRUNCATED AT 250 WORDS)
Bonnekoh
B
B
Department of Dermatology, University of Cologne, F.R.G.
Daefler
S
S
Krueger
G R
GR
Mahrle
G
G
D000328:Adult; D000818:Animals; D002478:Cells, Cultured; D002784:Cholesterol / Q000494:pharmacology*; D002788:Cholesterol Esters / Q000494:pharmacology; D006168:Guinea Pigs; D006801:Humans; D006854:Hydrocortisone / Q000494:pharmacology*; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D008560:Membrane Fluidity / Q000187:drug effects*; D012871:Skin Diseases / Q000378:metabolism* / Q000473:pathology; D013050:Spectrometry, Fluorescence / Q000379:methods; D014212:Tretinoin / Q000494:pharmacology*; D014810:Vitamin E / Q000494:pharmacology*
D002788:Cholesterol Esters; D014810:Vitamin E; D014212:Tretinoin; D002784:Cholesterol; C013440:cholesteryl succinate; D006854:Hydrocortisone