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目的观察长期高浓度氯化钠刺激对白假丝酵母菌耐受氧化应激能力的影响。方法选用2株白假丝酵母菌国际标准株SC5314和ATCC76615,用含有1mol/L氯化钠的YEPD培养液连续培养24d,考察子代与亲本菌对氧化应激的耐受能力。采用real-time RT-PCR考察白假丝酵母菌抗氧化酶的转录水平,并测定其抗氧化酶的活性,利用丙二醛(MDA)测定试剂盒衡量白假丝酵母菌脂质过氧化损伤,并测量白假丝酵母菌菌体细胞内活性氧水平。结果长期用高浓度氯化钠刺激能增加白假丝酵母菌对过氧化氢(12mmol/L)和咪康唑(4μg/ml)的耐受性,促进其抗氧化酶的转录,使抗氧化酶SOD1、CAT1、谷胱甘肽还原酶(GR,转录基因为TTR1)的转录水平升高2～5倍,增强抗氧化酶的活性(P<0.05),降低脂质过氧化损伤(P<0.01),减少菌体细胞内活性氧水平(P<0.001)。结论长期高浓度氯化钠刺激能够增强白假丝酵母菌耐受氧化应激的能力。
Objective To observe the effect of long-term high-concentration sodium chloride stimulation on Candida albicans tolerance to oxidative stress. Methods Two isolates of Candida albicans SC5314 and ATCC76615 were selected and cultured in YEPD medium containing 1 mol / L sodium chloride for 24 days. The tolerance to oxidative stress in progeny and their parental strains was investigated. Real-time RT-PCR was used to examine the transcriptional level of antioxidant enzymes of Candida albicans and the activity of antioxidant enzymes was measured. The lipid peroxidation injury of Candida albicans was evaluated by using MDA assay kit , And measured the level of reactive oxygen species in Candida albicans cells. Results Long-term stimulation with high concentration of sodium chloride increased the tolerance of Candida albicans to hydrogen peroxide (12 mmol / L) and miconazole (4 μg / ml), promoted the transcription of antioxidant enzymes, The transcription level of SOD1, CAT1 and glutathione reductase (GR, TTR1) were increased by 2 ~ 5 times, and the activity of antioxidant enzymes was increased (P <0.05) and the lipid peroxidation injury was decreased (P < 0.01), reduce the intracellular reactive oxygen species (P <0.001). Conclusion Long-term high-concentration sodium chloride stimulation can enhance the ability of Candida albicans to tolerate oxidative stress.