Limits...
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

Effects of SHI on cell protein expression. a Protein Expression in Kc; b Protein expression in HSF. Proteins were collected separately from Kc and HSF treated with SHI for 24 and 48 h. The expression of proteins was detected using the Odyssey Infrared Imaging system. GAPDH was included as a loading control. For quantitative analysis, the intensities of the bands were measured with densitometry and first normalized to GAPDH and then further converted to the percentage of the untreated control. The converted data from 5 different patients were pooled together as shown in the figure. Representative images of the western blots are presented. Error bars indicate mean +/− SEM (n = 5). *p < 0.05 versus the untreated control. Statistical analysis was performed using One-way ANOVA and Tukey’s post-hoc test
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig3: Effects of SHI on cell protein expression. a Protein Expression in Kc; b Protein expression in HSF. Proteins were collected separately from Kc and HSF treated with SHI for 24 and 48 h. The expression of proteins was detected using the Odyssey Infrared Imaging system. GAPDH was included as a loading control. For quantitative analysis, the intensities of the bands were measured with densitometry and first normalized to GAPDH and then further converted to the percentage of the untreated control. The converted data from 5 different patients were pooled together as shown in the figure. Representative images of the western blots are presented. Error bars indicate mean +/− SEM (n = 5). *p < 0.05 versus the untreated control. Statistical analysis was performed using One-way ANOVA and Tukey’s post-hoc test

Mentions: Mechanisms underlying the proliferation-inhibiting and apoptosis-inducing ability of SHI were determined using western blot approaches (Fig. 3). Mitogen-activated protein kinases (MAPK), including ERK1/2, JNK1/2 and p38α/β, have been widely demonstrated to play essential roles in cell proliferation and apoptosis [18]. Those kinases are activated by phosphorylation [19]. In addition, Bcl-2 can indirectly cleave caspase 3 by releasing cytochrome c from the mitochondria [20]. Cleaved caspase 3 will then further induce apoptosis [21]. As shown in Fig. 3a & b, increases in phosphorylated ERK1/2 and JNK1/2 (p-ERK1/2, p-JNK1/2), decreases in phosphorylated p38α/β (p-p38α/β) and Bcl-2, as well as cleavage of caspase 3, were observed in Kc at 48 h and in HSF at either 24 or 48 h after exposure to 3 μg/mL SHI compared to the untreated control (p < 0.05). These changes were also found in HSF when treated with 1 μg/mL SHI for 48 h, but no effect on Kc protein expression was detected at this dose of SHI. Taken together these results indicate that SHI increases p-ERK1/2 and p-JNK1/2, decreases p38α/β and Bcl-2 and induces cleavage of caspase 3 in both Kc and HSF in a dose-dependent manner. Again, these changes in protein expression occur more rapidly (24 h) in HSF than Kc at the same dose of SHI (3 μg/mL). SHI at 1 μg/mL can also trigger these changes in HSF but does not induce changes in these proteins in Kc.Fig. 3


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)

Effects of SHI on cell protein expression. a Protein Expression in Kc; b Protein expression in HSF. Proteins were collected separately from Kc and HSF treated with SHI for 24 and 48 h. The expression of proteins was detected using the Odyssey Infrared Imaging system. GAPDH was included as a loading control. For quantitative analysis, the intensities of the bands were measured with densitometry and first normalized to GAPDH and then further converted to the percentage of the untreated control. The converted data from 5 different patients were pooled together as shown in the figure. Representative images of the western blots are presented. Error bars indicate mean +/− SEM (n = 5). *p < 0.05 versus the untreated control. Statistical analysis was performed using One-way ANOVA and Tukey’s post-hoc test
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig3: Effects of SHI on cell protein expression. a Protein Expression in Kc; b Protein expression in HSF. Proteins were collected separately from Kc and HSF treated with SHI for 24 and 48 h. The expression of proteins was detected using the Odyssey Infrared Imaging system. GAPDH was included as a loading control. For quantitative analysis, the intensities of the bands were measured with densitometry and first normalized to GAPDH and then further converted to the percentage of the untreated control. The converted data from 5 different patients were pooled together as shown in the figure. Representative images of the western blots are presented. Error bars indicate mean +/− SEM (n = 5). *p < 0.05 versus the untreated control. Statistical analysis was performed using One-way ANOVA and Tukey’s post-hoc test
Mentions: Mechanisms underlying the proliferation-inhibiting and apoptosis-inducing ability of SHI were determined using western blot approaches (Fig. 3). Mitogen-activated protein kinases (MAPK), including ERK1/2, JNK1/2 and p38α/β, have been widely demonstrated to play essential roles in cell proliferation and apoptosis [18]. Those kinases are activated by phosphorylation [19]. In addition, Bcl-2 can indirectly cleave caspase 3 by releasing cytochrome c from the mitochondria [20]. Cleaved caspase 3 will then further induce apoptosis [21]. As shown in Fig. 3a & b, increases in phosphorylated ERK1/2 and JNK1/2 (p-ERK1/2, p-JNK1/2), decreases in phosphorylated p38α/β (p-p38α/β) and Bcl-2, as well as cleavage of caspase 3, were observed in Kc at 48 h and in HSF at either 24 or 48 h after exposure to 3 μg/mL SHI compared to the untreated control (p < 0.05). These changes were also found in HSF when treated with 1 μg/mL SHI for 48 h, but no effect on Kc protein expression was detected at this dose of SHI. Taken together these results indicate that SHI increases p-ERK1/2 and p-JNK1/2, decreases p38α/β and Bcl-2 and induces cleavage of caspase 3 in both Kc and HSF in a dose-dependent manner. Again, these changes in protein expression occur more rapidly (24 h) in HSF than Kc at the same dose of SHI (3 μg/mL). SHI at 1 μg/mL can also trigger these changes in HSF but does not induce changes in these proteins in Kc.Fig. 3

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