IDO1影响外阴阴道念珠菌病小鼠阴道组织中巨噬细胞功能变化的研究  

Effect of IDO1 on functional changes in macrophages in vaginal tissues from mouse models of vulvovaginal candidiasis

作　　者：唐诗琴 郝瑞英 贺慧娜 田亚楠 徐艳艳 李小静 景婷婷[2,3] Tang Shiqin;Hao Ruiying;He Huina;Tian Yanan;Xu Yanyan;Li Xiaojing;Jing Tingting(School of Clinical Medicine,Hebei University of Engineering,Handan 056002,Hebei,China;Department of Dermatology,Affiliated Hospital of Hebei University of Engineering,Handan 056002,Hebei,China;Hebei Immunological Dermatology Laboratory,Handan 056002,Hebei,China;Hebei University of Engineering,Handan 056038,Hebei,China)

机构地区：[1]河北工程大学临床医学院,邯郸056002 [2]河北工程大学附属医院皮肤科,邯郸056002 [3]河北省免疫性皮肤病实验室,邯郸056002 [4]河北工程大学,邯郸056038

出　　处：《中华皮肤科杂志》2024年第10期931-939,共9页Chinese Journal of Dermatology

基　　金：河北省自然科学基金(H2021402009);河北省临床医学优秀人才培养项目(ZF2023222、ZF2024205)。

摘　　要：目的通过调控吲哚胺2,3-双加氧酶(IDO1)观察外阴阴道念珠菌病(VVC)小鼠模型中巨噬细胞功能的变化。方法40只无特定病原级雌性ICR小鼠按完全随机法分为4组:空白组、VVC模型组、VVC模型+1-甲基色氨酸组(简称1-MT组)、VVC模型+γ干扰素组(简称IFN-γ组),每组10只。除空白组外,在造模前6 d,每隔1天予小鼠腹部皮下注射苯甲酸雌二醇油剂诱导假发情;发情成功后开始建模,继续皮下注射苯甲酸雌二醇油剂,予小鼠阴道接种2×109 CFU/ml白念珠菌菌悬液,每天1次,连续5 d;1-MT组和IFN-γ组分别在接种菌悬液前1 d予1-MT或IFN-γ预处理,之后每天1次,连续干预6 d;空白组和VVC模型组给予相应的生理氯化钠溶液。在造模第5天时,观察小鼠阴道情况,并进行阴道症状评分;收集阴道灌洗液涂片镜检和真菌载荷计数;随后处死小鼠,收集阴道组织,分别进行苏木精-伊红(HE)染色及过碘酸希夫(PAS)染色,免疫荧光法检测组织中IDO1和巨噬细胞表面标志物F4/80的表达;实时荧光定量PCR(qPCR)检测组织中IDO1、诱导型一氧化氮合酶(iNOS)、精氨酸酶1(Arg-1)mRNA表达水平,Western印迹检测组织中IDO1蛋白表达。采用单因素方差分析检验组间各指标的差异,Tukey检验进行两两多重比较。结果造模第5天,小鼠阴道灌洗液涂片镜检,VVC模型组中菌丝细长,见少量孢子;1-MT组大量细长菌丝聚集成团,周围可见孢子;IFN-γ组见少量细短菌丝,孢子较多。小鼠阴道灌洗液载菌量计数显示,与VVC模型组载菌量(360.0±15.9)相比,1-MT组较高(523.7±67.7,P=0.002),而IFN-γ组较低(258.3±27.6,P=0.026)。HE染色和PAS染色显示,VVC模型组表皮局灶可见小脓肿形成,表皮及真皮全层见以中性粒细胞为主的炎症细胞浸润,且存在大量孢子,伴少量菌丝;1-MT组表皮增生、增厚,真皮可见大量以中性粒细胞为主的炎症细胞弥漫性浸润,表皮及真皮全层大量菌丝和孢子聚集成团,以菌Objective To analyze functional changes in macrophages in mouse models of vulvovaginal candidiasis(VVC)by modulating indoleamine 2,3-dioxygenase 1(IDO1).Methods Forty specific-pathogen-free female ICR mice were randomly divided into 4 groups using a complete randomization method:a blank group,a VVC model group,a VVC model+1-methyltryptophan(1-MT)group(referred to as the 1-MT group),a VVC model+interferon-γ(IFN-γ)group(referred to as the IFN-γgroup),with 10 mice in each group.Except for the blank group,all the mice were injected subcutaneously with estradiol benzoate oil solution in the abdomen every other day for 6 days prior to modeling to induce pseudoestrus;after successful induction of pseudoestrus,the mice were inoculated vaginally with Candida albicans suspensions at a concentration of 2×109 CFU/ml once a day for 5 days to establish VVC mouse models,and subcutaneous injections of estradiol benzoate oil solution were continued simultaneously to maintain the pseudoestrus state;1 day before inoculation with fungal suspensions,mice in the 1-MT group and IFN-γgroup were pretreated with 1-MT and IFN-γrespectively,followed by once-daily same intervention for 6 consecutive days;mice in the blank group and VVC model group were intraperitoneally injected with physiological saline solution once a day for 6 consecutive days.On the 5th day of modeling,vaginal conditions in mice were observed and vaginal symptoms were scored;the vaginal lavage fluid was collected for smear microscopy and fungal colony counting;then,the mice were sacrificed,the vaginal tissues were collected and subjected to hematoxylin-eosin(HE)staining and periodic acid-Schiff(PAS)staining,and the expression of IDO1 and the macrophage surface marker F4/80 was determined in the vaginal tissues by an immunofluorescence method;real-time fluorescence-based quantitative PCR(qPCR)was performed to determine mRNA expression levels of IDO1,inducible nitric oxide synthase(iNOS),and arginase 1(Arg-1)in the vaginal tissues,and Western blot analysis to determi

关 键 词：白色念珠菌 念珠菌病 外阴阴道 巨噬细胞 吲哚胺2 3-双加氧酶 宿主免疫 

分 类 号：R711.31[医药卫生—妇产科学]