Induction of Apoptosis in Candida albicans by Semi-purified Fraction of Hypnea musciformis

Mary Santhi R
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Abstract

The leading cause of death worldwide is microbial infections. Amidst the rise of new species of Candida , Candida albicans leads the list with high morbidity and mortality rates. One of the most opportunistic pathogens responsible for infectious diseases in human beings is Candida albicans . To overcome this problem, there is a need to explore new antifungals that could replace the current treatment options. Marine red alga Hypnea musciformis was collected from the coastal areas of Rameswaram. The collected alga was cleaned, shade-dried, and powdered and extraction was done using the Soxhlet apparatus. The crude methanolic extract was subjected to bioassay-guided silica gel chromatography. A total of six fractions was obtained, among the six fractions, the fifth fraction was subjected to time killing assay, biofilm reduction assay and phosphatidylserine externalization against Candida albicans . The aim of the current study was to evaluate the potential of a semi-purified fraction of Hypnea musciformis to induce apoptosis in Candida albicans. In time killing assay, 1× and 2× MIC concentrations within 4 to 6 hr, the fungal cells were eradicated rapidly. The corresponding MIC values are required to reduce 50% of the biofilm’s metabolic activity 30-2000 times. From the results, it was evident that the SPF of H. musciformis exhibited good antibiofilm activity. SPF induced the production and accumulation of intracellular ROS, which finally induces apoptotic features in C. albicans, was confirmed using Annexin V double staining assay.
半纯化白丝酵母菌诱导白色念珠菌凋亡的研究
微生物感染是全世界死亡的主要原因。在念珠菌新种的兴起中,白色念珠菌以其高发病率和死亡率居首位。白色念珠菌是人类传染性疾病最具机会性的病原体之一。为了克服这个问题,有必要探索新的抗真菌药物,以取代目前的治疗方案。在Rameswaram沿海地区采集了海红藻Hypnea musciformis。收集的藻类经清洗、遮荫干燥,用索氏装置进行粉末和提取。对粗甲醇提取物进行生物测定导向硅胶层析。共得到6个组分,其中第5组分对白色念珠菌进行时间杀伤试验、生物膜还原试验和磷脂酰丝氨酸外化试验。当前研究的目的是评估半纯化的Hypnea musciformis部分诱导白色念珠菌凋亡的潜力。在时间杀伤实验中,1倍和2倍MIC浓度在4 ~ 6小时内,真菌细胞被迅速根除。相应的MIC值需要使生物膜的代谢活性降低50% 30-2000倍。结果表明,该菌具有良好的抗膜活性。通过Annexin V双染色法证实了SPF诱导细胞内ROS的产生和积累,最终诱导白色念珠菌的凋亡特征。
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