Abstract
Purpose : To investigate the effect of extracts of Artemisia anomala S. Moore tissues on viability, apoptosis and antioxidant capacity of human keratinocytes. Methods : Human keratinocyte cell line HaCaT were treated with extracts of A. anomala for 12 h or 24 h. Cell viability, level of reactive oxygen species (ROS), and incidence of apoptosis were measured by flow cytometry. Levels of mRNA and key proteins in the mitogen-activated protein kinase (MAPK) pathway were determined by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Key proteins of caspase pathways were assessed by western blot. The influence of the extract on the MAPK pathway was further probed by treating cells with MAPK activator in the presence and absence of the extract. Results : Treatment of cells with extracts of A. anomala enhanced viability and reduced apoptosis in a time-dependent manner, and increased ROS level, compared with control. mRNA and protein expressions of c-Jun N-terminal kinase (JNK), extracellular regulated protein kinase (ERK), and p38 MAPK decreased in extract-treated cells. The extracts also reversed the inhibitory effects of the MAPK pathway activator, actinomycin, on cell viability and ROS, and inhibited protein-cleaved caspase-8 and cleaved caspase-3. Conclusion : A. anomala extract increases cell viability and antioxidant capacity via inactivation of MAPK pathway, and also inhibits cell apoptosis via inactivation of caspase pathways. Hence, the extract may serve as a promising drug for the treatment of psoriasis. Keywords : Artemisia anomala , MAPK pathway, Anti-oxidation, Keratinocyte, Psoriasis
Highlights
The present study was undertaken to determine the effect of A. anomala extracts on cell viability and apoptosis of keratinocytes, as well as the antioxidant properties of keratinocytes treated with these extracts
Reverse transcription was performed with the RevertAid First Strand cDNA Synthesis Kit (Fermentas, USA) using 2 μg of RNA. quantitative reverse-transcription polymerase chain reaction (qRT-PCR) for c-Jun N-terminal kinase (JNK), extracellular regulated protein kinase (ERK), and mitogen-activated protein kinase (MAPK) was carried out via FastStart Universal SYBR Green Master (Roche Diagnostics, Tokyo, Japan)
The effect of A. anomala extracts on HaCaT cell growth were assessed 12 or 24 h after treatment by MTT assay
Summary
The present study was undertaken to determine the effect of A. anomala extracts on cell viability and apoptosis of keratinocytes, as well as the antioxidant properties of keratinocytes treated with these extracts. The keratinocyte cell line HaCaT (American Type Culture Collection, Manassas, VA, USA) was used. The effect of the extract on the HaCaT keratinocytes in media (DMEM) was evaluated For this experiment, HaCaT cells were incubated in A. anomala extract for 12 h or h, respectively. Keratinocytes treated with different extracts were placed in 6-well culture plates, fixed with 2% paraformaldehyde, washed with cold PBS, and re-suspended in 100 μL annexin-binding buffer. At this point, cell counting was carried out.
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