Cytotoxicity of Capsaicin on Cultured Human Skin Fibroblast.
- Author:
Seok Kweon YUN
1
;
Jong Keun KIM
;
Seong Jin KIM
;
Young Ho WON
Author Information
1. Department of Dermatology, Chonbuk National University, Korea.
- Publication Type:Original Article
- Keywords:
Capsaicin;
Cytotoxicity;
Fibroblast
- MeSH:
Antioxidants;
Ascorbic Acid;
Blotting, Western;
Calcium;
Capsaicin*;
Caspase 3;
Cell Survival;
Chromatin;
Cycloheximide;
Cytoplasm;
DNA;
Epidermal Growth Factor;
Fibroblasts*;
Humans*;
In Situ Nick-End Labeling;
Intercellular Signaling Peptides and Proteins;
Neuropeptides;
Ruthenium Red;
Skin*;
Substance P;
Tyrosine
- From:Korean Journal of Dermatology
2005;43(2):194-203
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: Capsaicin has been shown to have different biologic and toxic effects, depending on non-neuronal cells and several transformed cells, however no study has been reported from cultured human skin fibroblast. OBJECTIVE: Present study was aimed to evaluate the cytotoxicity and its mechanism of capsacin on the cultured human skin fibroblast. MATERIAL AND METHOD: Normal neonatal human fibroblasts were used, and changes of cell survival were measured by MTT assay after the cells were pre-treated with growth factors, receptor antagonist, antioxidants, calcium modulators were pre-treated or co-treated with capsaicin. RESULTS: Suvival of fibroblast was significantly increased by treatment with EGF (10ng/ml), bFGF (10ng/ml), capsazepine (10M) but inhibited by cycloheximide (1g/ml). When 200 M capsaicin was added to fibroblasts, chromatin condensations were observed at 12 hours and cell survival rate was reduced to 25-50% at 24 hours. Vanilloid receptor antagonists, capsazepine and ruthenium red, did not prevent the toxic effect of capsaicin, and 10M capsazepine paradoxically rather enhanced the cytotoxicity. In contrast to bFGF (10ng/ml), EGF (10, 100ng/ml) enhanced the cytotoxicity of capsaicin. Neuropeptides, substance P (1, 10nM) and CGRP (1, 10nM), and a structural analogue to capsaicin, tyrosine (0.3-1.2mM) did not affect the cytotoxicity. However, antioxidants such as trolox (100M) and ascorbic acid (0.1, 0.3 mM) reduced the capsaicin cytotoxicity. Of calcium modulating agents, nifedifine, a Ca2+ channel blocker (10, 20M) and cyclopiazonic acid, a Ca2+-ATPase inhibitor in ER (10M) did not influence the cytotoxicity, however BAPTA/AM (10M) as a chelater for cytoplasmic free calcium ion (10M) significantly decreased capsaicin cytotoxicity. Unlike cycloheximide, z-VAD-FMK, a protein synthesis inhibitor and a non-specific caspase inhibitor, prevented the capsaicin cytotoxicity. The DNA ladder and TUNEL positive cells were observed among the capsaicin treated fibroblasts and Western blot revealed caspase-3 activity. CONCLUSION: The capsaicin-induced cytotoxicity on human skin fibroblasts is likely to suggest the mechanism of an apoptotic pathway, which can possibly be prevented by antioxidants.
