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Investigating the potential of Shikonin as a novel hypertrophic scar treatment.

Fan C, Xie Y, Dong Y, Su Y, Upton Z - J. Biomed. Sci. (2015)

Bottom Line: Our results indicate that Shikonin preferentially inhibits cell proliferation and induces apoptosis in fibroblasts without affecting keratinocyte function.In addition, we found that the proliferation-inhibiting and apoptosis-inducing abilities of SHI might be triggered via MAPK and Bcl-2/Caspase 3 signalling pathways.Furthermore, SHI has been found to attenuate the expression of TGF-β1 in Transwell co-cultured "conditioned" medium.

View Article: PubMed Central - PubMed

Affiliation: Tissue Repair and Regeneration Program, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Queensland, 4059, Australia. c3.fan@connect.qut.edu.au.

ABSTRACT

Background: Hypertrophic scarring is a highly prevalent condition clinically and results from a decreased number of apoptotic fibroblasts and over-abundant production of collagen during scar formation following wound healing. Our previous studies indicated that Shikonin, an active component extracted from Radix Arnebiae, induces apoptosis and reduces collagen production in hypertrophic scar-derived fibroblasts. In the study reported here, we further evaluate the potential use of Shikonin as a novel scar remediation therapy by examining the effects of Shikonin on both keratinocytes and fibroblasts using Transwell® co-culture techniques. The underlying mechanisms were also revealed. In addition, effects of Shikonin on the expression of cytokines in Transwell co-culture "conditioned" medium were investigated.

Results: Our results indicate that Shikonin preferentially inhibits cell proliferation and induces apoptosis in fibroblasts without affecting keratinocyte function. In addition, we found that the proliferation-inhibiting and apoptosis-inducing abilities of SHI might be triggered via MAPK and Bcl-2/Caspase 3 signalling pathways. Furthermore, SHI has been found to attenuate the expression of TGF-β1 in Transwell co-cultured "conditioned" medium.

Conclusions: The data generated from this study provides further evidence that supports the potential use of Shikonin as a novel scar remediation therapy.

No MeSH data available.


Related in: MedlinePlus

Link between MAPK and intrinsic apoptosis signalling pathway. HSF were treated with SHI with or without U0126 (10 μM) or SP600125 (50 μM) for 24 h. Expression of protein was measured using the Odyssey Infrared Imaging system. All experiments were performed 3 times using cells from 3 patients. Triplicate treatments were assessed in cells from each patient. The data was pooled as the percentage of the untreated control. Representative images of the western blots are presented. Quantitative data were pooled from experiments using cells 3 different patients. *p < 0.05
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Fig4: Link between MAPK and intrinsic apoptosis signalling pathway. HSF were treated with SHI with or without U0126 (10 μM) or SP600125 (50 μM) for 24 h. Expression of protein was measured using the Odyssey Infrared Imaging system. All experiments were performed 3 times using cells from 3 patients. Triplicate treatments were assessed in cells from each patient. The data was pooled as the percentage of the untreated control. Representative images of the western blots are presented. Quantitative data were pooled from experiments using cells 3 different patients. *p < 0.05

Mentions: As described above, we demonstrated that SHI up-regulates p-ERK and p-JNK and down-regulates p-p38, caspase 3 and Bcl-2 expression in Kc and HSF in a dose-dependent manner. Reports in the literature indicate that the MAPK proteins play roles in regulating cell apoptosis processes [22, 23]. To investigate the roles of MAPK proteins in SHI-induced apoptosis, the p-ERK inhibitor U0126 and the p-JNK inhibitor SP600125 were used to block the phosphorylation of ERK and JNK in HSF when treated with SHI. The results of this analysis (Fig. 4) indicated that U0126 significantly inhibits SHI-induced up-regulation of p-ERK in HSF at 24 h. When HSF were treated with both SHI (1 and 3 μg/mL) and U0126, there was no reduction in Bcl-2 and cleaved caspase 3 was observed, indicating that the blockage of p-ERK interrupts SHI-induced down-regulation of Bcl-2 and cleavage of caspase 3. However, reductions in Bcl-2 and cleavage of caspase 3 were detected when HSF were treated with both SHI and SP600125, indicating that blockage of p-JNK has no effect on SHI-induced apoptosis in HSF.Fig. 4


Investigating the potential of Shikonin as a novel hypertrophic scar treatment.

Fan C, Xie Y, Dong Y, Su Y, Upton Z - J. Biomed. Sci. (2015)

Link between MAPK and intrinsic apoptosis signalling pathway. HSF were treated with SHI with or without U0126 (10 μM) or SP600125 (50 μM) for 24 h. Expression of protein was measured using the Odyssey Infrared Imaging system. All experiments were performed 3 times using cells from 3 patients. Triplicate treatments were assessed in cells from each patient. The data was pooled as the percentage of the untreated control. Representative images of the western blots are presented. Quantitative data were pooled from experiments using cells 3 different patients. *p < 0.05
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
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getmorefigures.php?uid=PMC4537585&req=5

Fig4: Link between MAPK and intrinsic apoptosis signalling pathway. HSF were treated with SHI with or without U0126 (10 μM) or SP600125 (50 μM) for 24 h. Expression of protein was measured using the Odyssey Infrared Imaging system. All experiments were performed 3 times using cells from 3 patients. Triplicate treatments were assessed in cells from each patient. The data was pooled as the percentage of the untreated control. Representative images of the western blots are presented. Quantitative data were pooled from experiments using cells 3 different patients. *p < 0.05
Mentions: As described above, we demonstrated that SHI up-regulates p-ERK and p-JNK and down-regulates p-p38, caspase 3 and Bcl-2 expression in Kc and HSF in a dose-dependent manner. Reports in the literature indicate that the MAPK proteins play roles in regulating cell apoptosis processes [22, 23]. To investigate the roles of MAPK proteins in SHI-induced apoptosis, the p-ERK inhibitor U0126 and the p-JNK inhibitor SP600125 were used to block the phosphorylation of ERK and JNK in HSF when treated with SHI. The results of this analysis (Fig. 4) indicated that U0126 significantly inhibits SHI-induced up-regulation of p-ERK in HSF at 24 h. When HSF were treated with both SHI (1 and 3 μg/mL) and U0126, there was no reduction in Bcl-2 and cleaved caspase 3 was observed, indicating that the blockage of p-ERK interrupts SHI-induced down-regulation of Bcl-2 and cleavage of caspase 3. However, reductions in Bcl-2 and cleavage of caspase 3 were detected when HSF were treated with both SHI and SP600125, indicating that blockage of p-JNK has no effect on SHI-induced apoptosis in HSF.Fig. 4

Bottom Line: Our results indicate that Shikonin preferentially inhibits cell proliferation and induces apoptosis in fibroblasts without affecting keratinocyte function.In addition, we found that the proliferation-inhibiting and apoptosis-inducing abilities of SHI might be triggered via MAPK and Bcl-2/Caspase 3 signalling pathways.Furthermore, SHI has been found to attenuate the expression of TGF-β1 in Transwell co-cultured "conditioned" medium.

View Article: PubMed Central - PubMed

Affiliation: Tissue Repair and Regeneration Program, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Queensland, 4059, Australia. c3.fan@connect.qut.edu.au.

ABSTRACT

Background: Hypertrophic scarring is a highly prevalent condition clinically and results from a decreased number of apoptotic fibroblasts and over-abundant production of collagen during scar formation following wound healing. Our previous studies indicated that Shikonin, an active component extracted from Radix Arnebiae, induces apoptosis and reduces collagen production in hypertrophic scar-derived fibroblasts. In the study reported here, we further evaluate the potential use of Shikonin as a novel scar remediation therapy by examining the effects of Shikonin on both keratinocytes and fibroblasts using Transwell® co-culture techniques. The underlying mechanisms were also revealed. In addition, effects of Shikonin on the expression of cytokines in Transwell co-culture "conditioned" medium were investigated.

Results: Our results indicate that Shikonin preferentially inhibits cell proliferation and induces apoptosis in fibroblasts without affecting keratinocyte function. In addition, we found that the proliferation-inhibiting and apoptosis-inducing abilities of SHI might be triggered via MAPK and Bcl-2/Caspase 3 signalling pathways. Furthermore, SHI has been found to attenuate the expression of TGF-β1 in Transwell co-cultured "conditioned" medium.

Conclusions: The data generated from this study provides further evidence that supports the potential use of Shikonin as a novel scar remediation therapy.

No MeSH data available.


Related in: MedlinePlus