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In Vitro Effect of Thai herbal extracts with anti-psoriatic activity on the expression of caspase 9

In Vitro Effect of Thai herbal extracts with anti-psoriatic activity on the expression of caspase 9

Psoriasis, known as the incurable disease, is a chronic inflammatory disease characterize by hyperproliferative keratinocytes. The causes of this disease are unclear, and many side effects are evident as a result of certain treatments of psoriasis. In this work, we aimed at studying the in vitro effect of each of these following Thai herbal extracts, Alpinia galanga L. (rhizome), Curcuma longa L. (rhizome) and Annona squamosa L.(leaf), containing anti-psoriatic activity on the expression of caspase 9, the protein known to play a critical role in the intrinsic apoptotic pathway. The HaCaT keratinocyte cell line was chosen as an in vitro model. Based on Western blot analysis, the extract derived from Annona squamosa L. leaf at all three concentrations tested significantly reduced the expression of procaspase 9 (P<0.05), and two higher concentrations, 0.5IC50 (3.15 µg/mL) and IC50 (6.3 µg/mL), of this herbal extract significantly increased the expression of caspase 9 protien (P<0.05). Confocal immunofluorescence microscopy subsequently confirmed this finding. Taken together, this might suggest that the ethanolic extract of Annona squamosa leaf with anti-psoriatic activity could exert its biological effect by activating the cleavage of rocaspase 9 into caspase 9, thus inducing the intrinsic apoptotic pathway.

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98 views • 8 slides
Effects of ethanolic extract of Annona squamosa L. leaves on the expression of EGFR

Effects of ethanolic extract of Annona squamosa L. leaves on the expression of EGFR

The precise causes of psoriasis, a relatively common, chronic, inflammatory and hyperproliferative skin disease, are still not known, thus making it very difficult for treatment. We previously found an in vitro anti-psoriatic activity in ethanolic extract of Annona squamosa L. leaves. Epidermal growth factor receptor (EGFR) and its ligand, transforming growth factor (TGF)-α, have been demonstrated to be strikingly upregulated in active psoriatic plaques, thus implicating their functional roles in psoriatic hyperplasia. The objective of this study was to investigate the molecular effect of ethanolic extract derived from Annona squamosa L. leaves on the EGFR expression using HaCaT keratinocyte cell line as a model. Based on RT-PCR, concentrations at 3.15 and 1.575 µg/mL (IC50= 6.3 µg/mL) significantly reduced the EGFR mRNA expression (P<0.05) as compared to control (HaCaT pretreated with IFN-γ and TNF-α, 10 ng/mL for each) although all three concentrations tested showed a similar tendency of inhibitory effect. Using confocal immunofluorescence microscopy as a qualitative measurement, we also found the inhibitory effect of Annona squamosa L. leaf extract on the expression of EGFR protein. Western blot analysis confirmed that all concentrations of the extract tested markedly inhibited the expression of EGFR (P<0.05). Taken together, this suggest that the ethanolic extract of Annona squamosa leaves could exert its biological effect by suppressing the expression of EGFR biomarker, thus highlighting the potential therapeutic development of this extract for future applications.

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90 views • 7 slides
Effects of ethanolic extract of Annona squamosa L. leaves on the expression of EGFR

Effects of ethanolic extract of Annona squamosa L. leaves on the expression of EGFR

The precise causes of psoriasis, a relatively common, chronic, inflammatory and hyperproliferative skin disease, are still not known, thus making it very difficult for treatment. We previously found an in vitro anti-psoriatic activity in ethanolic extract of Annona squamosa L. leaves. Epidermal growth factor receptor (EGFR) and its ligand, transforming growth factor (TGF)-α, have been demonstrated to be strikingly upregulated in active psoriatic plaques, thus implicating their functional roles in psoriatic hyperplasia. The objective of this study was to investigate the molecular effect of ethanolic extract derived from Annona squamosa L. leaves on the EGFR expression using HaCaT keratinocyte cell line as a model. Based on RT-PCR, concentrations at 3.15 and 1.575 µg/mL (IC50= 6.3 µg/mL) significantly reduced the EGFR mRNA expression (P<0.05) as compared to control (HaCaT pretreated with IFN-γ and TNF-α, 10 ng/mL for each) although all three concentrations tested showed a similar tendency of inhibitory effect. Using confocal immunofluorescence microscopy as a qualitative measurement, we also found the inhibitory effect of Annona squamosa L. leaf extract on the expression of EGFR protein. Western blot analysis confirmed that all concentrations of the extract tested markedly inhibited the expression of EGFR (P<0.05). Taken together, this suggest that the ethanolic extract of Annona squamosa leaves could exert its biological effect by suppressing the expression of EGFR biomarker, thus highlighting the potential therapeutic development of this extract for future applications.

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93 views • 7 slides

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