δ氨基酮戊酸光动力学疗法用于皮肤Fusarium equiseti感染的治疗及其机制  

Treatment and mechanism of delta-aminolevulinic acid mediated photodynamic therapy in cutaneous Fusarium equiseti infection

作　　者：梁嫔 孙毅[2] 蒋思[1] 郑楷平 彭黎慧 易文娟 LIANG Pin;SUN Yi;JIANG Si;ZHENG Kaiping;PENG Lihui;YI Wenjuan(Dept,of Dermatology,Zhongnan Hospital of Wuhan University.Wuhan 430071,Hubei,China;Dept.of Dermatology,Jingzhou Center Hospital,Jifigzhou 434020,Hubei,China)

机构地区：[1]武汉大学中南医院皮肤科,湖北武汉430071 [2]荆州市中心医院皮肤科,湖北荆州434020

出　　处：《武汉大学学报（医学版）》2021年第1期103-107,共5页Medical Journal of Wuhan University

基　　金：国家自然科学基金资助项目(编号:81602771)。

摘　　要：目的:报道1例由Fusarium equiseti(F.equiseti)所致的皮肤镰刀菌感染,探讨δ氨基酮戊酸(ALA)光动力学疗法(PDT)对镰刀菌杀伤效应及作用机制。方法:取皮损组织培养物进行菌种鉴定,体外培养后制备菌悬液,使其与不同浓度ALA(2.5%、5%)悬浮培养,置于光动力治疗仪下光照,通过菌落计数法观察菌株的生长情况。使用Annexin V-Alexa Fluor 488/PI kit对上述处理组中的镰刀菌进行凋亡检测。结果:根据真菌培养物形态、PCR扩增ITS(ribosomal internal transcribed spacer)区序列、Calmodulin序列及TEF-1a分析,鉴定该真菌为F.equiseti,ALA-PDT对F.equiseti的杀伤与ALA浓度成正相关(P<0.05),单纯激光和只加ALA无激光光照均对真菌生长无明显抑制作用;且ALA-PDT可诱导该真菌的凋亡及坏死。结论:δ氨基酮戊酸介导的光动力学疗法对体外培养的F.equiseti有明确杀伤效应,为ALA-PDT治疗皮肤镰刀菌感染提供新的视角。Objective:To report a case of cutaneous infection caused by Fusarium equiseti(F.equiseti) and related laboratory investigation,then to evaluate the killing effect of delta-aminolevulinic acid(ALA)-mediated photodynamic therapy on F.equiseti in vitro and investigate its mechanism.Methods:Fungal culture,PCR for ribosomal internal transcribed spacer(ITS),calmodulin,and TEF-1 a sequences analysis were performed for the diagnosis of fungal strain,and the fungi strain were inoculated in Sabouraud glucose peptone,for preparing fungi suspensions.The suspensions were treated with different concentrations of ALA(2.5% and 5% respectively) cultures,and lighted up under photodynamic therapeutic apparatus.The colony forming units were used to examine the viability of fungi.The apoptosis rates of fungal conidia were observed by flow cytometry after the treatment of(2.5% or 5%) ALAPDT,with Annexin V-Alexa Fluor 488/PI kit.Results:The colony was found in fungal culture.Sickle type conidia were observed in the slide culture.Ribosomal DNA ITS domains,calmodulin and TEF-1 a sequences were amplified,and the products were sequenced,and were aligned with 100% sequential similarity to Fusarium equiseti in the GenBank database.After incubated with ALA,we demonstrated that there was a positive correlation between the concentration of ALA and the inhibition ratio of fungi(P<0.05),but simple laser and simple ALA treatment had no significant inhibition on fungal growth.And ALA-PDT can induce the apoptosis and necrosis of Fusarium.Conclusion:ALA-PDT could inhibit F.equiseti excellently in vitro,might through inducing fungal cell necrosis,which provides a theoretical basis for the treatment of cutaneous infection caused by F.equiseti.

关 键 词：Fusarium equiseti Δ-氨基酮戊酸 光动力学治疗 凋亡 

分 类 号：Q939.5[生物学—微生物学]