Abstract
Purpose: To determine the efficacy of Helichrysum petiolare ethanol leaf extract against skin aging.Methods: The cytotoxic potential of the plant extract towards human dermal fibroblast (MRHF) cells was determined by Hoechst 33342/Propidium iodide (PI) staining. Effect of H. petiolare extract on reactive oxygen species (ROS) levels in MRHF cells and NO (nitric oxide) production in RAW 246.7 cells activated by LPS (lipopolysaccharides) was investigated. The inhibitory effect of the extract againstcollagenase, elastase, tyrosinase and protein glycation was also evaluated.Results: The extract did not display cytotoxicity towards MRHF cells at the tested concentrations when compared to the trend seen with the untreated control (p < 0.05). The extract caused a significant decrease (p < 0.05) in ROS levels in MRHF cells in a concentration-dependent manner and also demonstrated a reduction in NO production in RAW cells with no toxicity. Furthermore, the extract produced a weak inhibition of collagenase, elastase and tyrosinase activities when compared to the corresponding positive controls, but effectively inhibited protein glycation at the tested concentrations.Conclusion: The findings suggest that the ethanol leaf extract from H. petiolare has the potential to mitigate skin aging and therefore needs to be further investigated for possible clinical applications.
 Keywords: Cytotoxicity, Efficacy, Helichrysum petiolare, MRHF cells, Oxidative stress, Protein glycation; skin aging
Highlights
Skin diseases have recently been of major health challenges worldwide and they amount to about 34% of all occupational diseases encountered [1]
The cytotoxic result obtained from the Hoechst/Propidium iodide (PI) dual staining method revealed that H. petiolare extract was not toxic to MRHF cells at all the tested concentrations, as evidenced by the trend seen in the percentage of the live cells (Figure 1)
Treatment of the cells with positive control, melphalan caused a significant decrease in the number of live cells when compared to that of extract and untreated control
Summary
Skin diseases have recently been of major health challenges worldwide and they amount to about 34% of all occupational diseases encountered [1]. The treatment medium was removed and replaced with 50 μL of staining solution (5 mL binding buffer (phosphate buffer saline with Ca2+ and Mg2+) containing Annexin V-FITC reagent (50 μL) and 2 μL Hoechst 33342 solution (10 mg/mL)). 20 μL substrate solution (4.4 mM succinyl-Ala-Ala-Alap-nitroanilide in Tris-HCl buffer) was added to the mixture to start the reaction and the absorbance was read at 410 nm for 10 min at 1 min intervals. The reaction mixture contained 20 μL extract (25- 200 μg/mL) or Kojic acid and blank (phosphate- buffered saline, pH 6.8) were pipetted into the 96-well plate. The reaction mixture contained 10 μL extract (50 - 200 μg/mL) or EDTA (0.5 mM) or catechin (0.5 mM), 10 μL collagenase enzyme solution and 10 μL gelatin. RAW cells was not as a result of cell death
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