Honokiol induces reactive oxygen species-mediated apoptosis in Candida albicans through mitochondrial dysfunctionReportar como inadecuado




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Objective

To investigate the effects of honokiol on induction of reactive oxygen species ROS, antioxidant defense systems, mitochondrial dysfunction, and apoptosis in Candida albicans.

Methods

To measure ROS accumulation, 2′,7′-dichlorofluorescein diacetate fluorescence was used. Lipid peroxidation was assessed using both fluorescence staining and a thiobarbituric acid reactive substances TBARS assay. Protein oxidation was determined using dinitrophenylhydrazine derivatization. Antioxidant enzymatic activities were measured using commercially available detection kits. Superoxide dismutase SOD genes expression was measured using real time RT-PCR. To assess its antifungal abilities and effectiveness on ROS accumulation, honokiol and the SOD inhibitor N,N′-diethyldithiocarbamate DDC were used simultaneously. Mitochondrial dysfunction was assessed by measuring the mitochondrial membrane potential mtΔψ. Honokiol-induced apoptosis was assessed using an Annexin V-FITC apoptosis detection kit.

Results

ROS, lipid peroxidation, and protein oxidation occurred in a dose-dependent manner in C. albicans after honokiol treatment. Honokiol caused an increase in antioxidant enzymatic activity. In addition, honokiol treatment induced SOD genes expression in C. albicans cells. Moreover, addition of DDC resulted in increased endogenous ROS levels and potentiated the antifungal activity of honokiol. Mitochondrial dysfunction was confirmed by measured changes to mtΔψ. The level of apoptosis increased in a dose-dependent manner after honokiol treatment.

Conclusions

Collectively, these results indicate that honokiol acts as a pro-oxidant in C. albicans. Furthermore, the SOD inhibitor DDC can be used to potentiate the activity of honokiol against C. albicans.



Autor: Lingmei Sun, Kai Liao, Chengcheng Hang, Dayong Wang

Fuente: http://plos.srce.hr/



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