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Aucklandia lappa Causes Membrane Permeation of Candida albicans
Heung-Shick Lee1, Younhee Kim2
1Department of Biotechnology and Bioinformatics, Korea University, Sejongsi 30019, Republic of Korea.
Abstract:
Candida albicans is a major fungal pathogen in humans. In our previous study, we reported that an ethanol extract from Aucklandia lappa weakens C. albicans cell wall by inhibiting synthesis or assembly of both (1,3)-β-D-glucan polymers and chitin. In the current study, we found that the extract is involved in permeabilization of C. albicans cell membranes. While uptake of ethidium bromide (EtBr) was 3.0% in control cells, it increased to 7.4% for 30 min in the presence of the A. lappa ethanol extract at its minimal inhibitory concentration (MIC), 0.78 mg/ml, compared to uptake by heat-killed cells. Besides, leakage of DNA and proteins was observed in A. lappa-treated C. albicans cells. The increased uptake of EtBr and leakage of cellular materials suggest that A. lappa ethanol extract induced functional changes in C. albicans cell membranes. Incorporation of diphenylhexatriene (DPH) into membranes in the A. lappa-treated C. albicans cells at its MIC decreased to 84.8%, after 60 min of incubation, compared with that of the controls, indicate that there was a change in membrane dynamics. Moreover, the anticandidal effect of the A. lappa ethanol extract was enhanced at a growth temperature of 40°C compared to that at 35°C. The above data suggest that the antifungal activity of the A. lappa ethanol extract against C. albicans is associated with synergistic action of membrane permeabilization due to changes in membrane dynamics and cell wall damage caused by reduced formation of (1,3)-β-D-glucan and chitin.
Insights
The Aucklandia lappa ethanol extract damages Candida albicans cell membranes and walls, inhibiting fungal growth. This extract shows enhanced antifungal activity, particularly at higher temperatures.
Area of Science:
- Mycology
- Pharmacology
- Biochemistry
Background:
- * Candida albicans is a significant human fungal pathogen.
- * Previous research indicated Aucklandia lappa extract weakens C. albicans cell walls by inhibiting glucan and chitin synthesis.
- * This study investigates the extract's effect on C. albicans cell membranes.
Purpose of the Study:
- * To determine if Aucklandia lappa ethanol extract affects C. albicans cell membrane integrity.
- * To elucidate the mechanism of the extract's antifungal activity.
- * To assess the influence of temperature on the extract's efficacy.
Main Methods:
- * Measured ethidium bromide (EtBr) uptake to assess membrane permeabilization.
- * Quantified leakage of DNA and proteins from treated cells.
- * Evaluated membrane dynamics using diphenylhexatriene (DPH) incorporation.
- * Compared antifungal activity at 35°C and 40°C.
Main Results:
- * Aucklandia lappa extract increased EtBr uptake and leakage of DNA/proteins, indicating membrane permeabilization.
- * DPH incorporation decreased in treated cells, suggesting altered membrane dynamics.
- * The extract's anticandidal effect was enhanced at 40°C compared to 35°C.
- * Minimal inhibitory concentration (MIC) of the extract was 0.78 mg/ml.
Conclusions:
- * Aucklandia lappa ethanol extract disrupts C. albicans cell membrane integrity and dynamics.
- * The extract exhibits synergistic antifungal activity through membrane permeabilization and cell wall damage.
- * Temperature influences the efficacy of the extract against C. albicans.
