The fungal cell wall is a major target of antifungals. In this study, we report the antifungal activity of an ethanol extract from Aucklandia lappa against Candida albicans. We found that the extract caused cell wall injury by decreasing chitin synthesis or assembly and (1,3)-β-D-glucan synth...
The fungal cell wall is a major target of antifungals. In this study, we report the antifungal activity of an ethanol extract from Aucklandia lappa against Candida albicans. We found that the extract caused cell wall injury by decreasing chitin synthesis or assembly and (1,3)-β-D-glucan synthesis. A sorbitol protection assay demonstrated that the minimum inhibitory concentration (MIC) of the A. lappa extract against C. albicans cells increased eight-fold from 0.78 to 6.24 mg/ml in 72 h. Cell aggregates, which indicate damage to the cell wall or membrane, were commonly observed in the A. lappatreated C. albicans cells through microscopic analysis. In addition, the relative fluorescence intensities of the C. albicans cells incubated with the A. lappa extract for 3, 5, and 6 h were 92.1, 84.6, and 79.8%, respectively, compared to the controls, estimated by Calcofluor White binding assay. This result indicates that chitin content was reduced by the A. lappa treatment. Furthermore, synthesis of (1,3)-β-D-glucan polymers was inhibited to 84.3, 79.7, and 70.2% of that of the control treatment following incubation of C. albicans microsomes with the A. lappa extract at a final concentration equal to its MIC, 2× MIC, and 4× MIC, respectively. These findings suggest that the A. lappa ethanol extract may aid the development of a new antifungal to successfully control Candidaassociated disease.
The fungal cell wall is a major target of antifungals. In this study, we report the antifungal activity of an ethanol extract from Aucklandia lappa against Candida albicans. We found that the extract caused cell wall injury by decreasing chitin synthesis or assembly and (1,3)-β-D-glucan synthesis. A sorbitol protection assay demonstrated that the minimum inhibitory concentration (MIC) of the A. lappa extract against C. albicans cells increased eight-fold from 0.78 to 6.24 mg/ml in 72 h. Cell aggregates, which indicate damage to the cell wall or membrane, were commonly observed in the A. lappatreated C. albicans cells through microscopic analysis. In addition, the relative fluorescence intensities of the C. albicans cells incubated with the A. lappa extract for 3, 5, and 6 h were 92.1, 84.6, and 79.8%, respectively, compared to the controls, estimated by Calcofluor White binding assay. This result indicates that chitin content was reduced by the A. lappa treatment. Furthermore, synthesis of (1,3)-β-D-glucan polymers was inhibited to 84.3, 79.7, and 70.2% of that of the control treatment following incubation of C. albicans microsomes with the A. lappa extract at a final concentration equal to its MIC, 2× MIC, and 4× MIC, respectively. These findings suggest that the A. lappa ethanol extract may aid the development of a new antifungal to successfully control Candidaassociated disease.
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문제 정의
tropicalis. This study aimed to reveal the mode of antifungal action against C. albicans, which is a major model organism among fungal pathogens.
가설 설정
albicans cells. Based on the data, it is hypothesized that the A. lappa-treated C. albicans cells have a thinner cell wall with loose chitin, and the treatment causes a series of events to take place such as the inhibition of the budding process and cell cycle arrest.
제안 방법
To investigate the effect of the A. lappa ethanol extract on the integrity of the C. albicans cell wall, a sorbitol protection assay was carried out using a modified CLSI M27-A3 protocol containing resazurin, as described above [15]. Briefly, two-fold serial dilutions of the A.
Damage of the essential cell wall components by antifungals will cause cell lysis, but cells can stay alive with an appropriate osmotic stabilizer in the medium [17]. To investigate whether the antifungal activity of A. lappa is linked to the modification of the fungal cell wall, CLSI M27-A3 microdilution assay was carried out with the A. lappa extract against C. albicans cells with or without 0.8 M sorbitol (Table 2). Regardless of the sorbitol, the MIC values of A.
대상 데이터
The standard strain of Candida albicans, SC5314, was purchased from the American Type Culture Collection (USA). C. glabrata (ATCC 2001, KCCM 50044), C. krusei (ATCC 32196, KCCM 11426), and C. tropicalis (ATCC 750, KCCM 50075), obtained from the Korean Culture Center of Microorganisms (KCCM), were included as controls.
데이터처리
The effect of A. lappa treatments when compared with controls was analyzed in SigmaPlot 13.0, using the Student’s t-test.
이론/모형
Antifungal susceptibilities of Candida species were determined by the modified CLSI M27-A3 method containing resazurin.
Antifungal susceptibility tests were carried out according to the modified CLSI M27-A3 protocol containing resazurin with or without 0.8 M sorbitol as an osmotic stabilizer. The 96-well round-bottom plate was incubated at 35°C, and minimum inhibitory concentrations (MICs) were determined after 24 and 72 h, respectively.
성능/효과
Synthesis of (1,3)-βD-glucan was inhibited to 84.3, 79.7, and 70.2% of that of the control treatment following treatment of microsomes with the A. lappa extract at a final concentration equal to its MIC, 2× MIC, and 4× MIC, respectively, and the differences between the DMSO control and the A. lappa-treated C. albicans cells were statistically significant (p < 0.01).
The relative fluorescence intensity of the A. lappa-treated cells decreased in a time-dependent manner, and the differences in fluorescence intensity between the DMSO control and each experimental group were statistically significant between 3 and 6 h (p < 0.05).
lappa ethanol extract decreases chitin content and inhibits (1,3)-β-D-glucan synthase activity. Therefore, it can be concluded that the A. lappa extract considerably damages the C. albicans cell wall and has therapeutic potential against Candida-associated infections.
albicans appeared thinner than those of the DMSO controls. Third, photobleaching, fading of the blue fluorescence of Calcofluor White, was more obvious in the A. lappa-treated C. albicans cells than in the controls. Hence, the thinner cell walls and remarkable photobleaching observed in the A.
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