摘要
目的探讨Th17／白细胞介素（IL）17轴在小鼠念珠菌性阴道炎发病中作用。方法120只雌性BALB／c小鼠随机分为雌激素感染组（Ei）、雌激素非感染组（En）、对照感染组（Ci）和对照非感染组（Cn）。Ei、En组在阴道接种前3d于后腿皮下注射雌二醇0．05mg，而Ci、Cn组皮下注射灭菌大豆油0．1ml，以后隔日注射1次直至实验结束；Ei、Ci组在接种日在阴道内接种白念珠菌悬液10μL（5×10^4个孢子），而En、Cn组阴道内注入10μl无菌磷酸盐缓冲液。接种后3、7、14d，每组于各时间点分别随机取10只小鼠处死，完整切取阴道组织作液氮冷冻或石蜡包埋，实时荧光定量PCR（qRT-PCR）、Western印迹和免疫荧光染色分别检测阴道组织中维A酸相关孤儿受体（ROR）γt、RORα、IL-17mRNA、蛋白表达及免疫荧光强度。结果激光共聚焦显微镜显示，En、Cn组中RORγt、RORα和IL—17主要表达于阴道固有膜及血管中炎症细胞，Ci组中主要表达于黏膜上皮及其表面附着的菌丝、固有膜及血管中炎症细胞，Ei组中则广泛分布于黏膜上皮、固有膜及血管中炎症细胞、阴道腔和黏膜上皮中内吞菌丝。qRT-PCR和免疫荧光检查显示，En、Ci和Ei组中RORγt、RORα、IL-17mRNA表达及其免疫荧光强度在相同时间点均显著高于Cn组（均P〈0．05），且Ei组显著高于En、ci组（P〈0．05）；除Cn组外，其余各组中RORγt、RORα、IL-17mRNA表达及其免疫荧光强度均随时间延长有增加的趋势，其中RORγt、RORαmRNA和免疫荧光强度及IL-17mRNA一般在14d时最高，而IL-17免疫荧光强度在7d时最高（P〈0．05）。Western印迹显示，Ci、Ei组中RORγt和IL-17蛋白表达在相同时间点均明显高于Cn、En组（RORγt：F组别=45．685，P〈0．001；IL-17：F组别=29．655，P〈0．01），其中H组表达水平最高（P〈0.05），而Cn、En组表达差异无统计学意义（P〉0．05）；Ci、Ei组中�
Objective To investigate the role of T helper 17 cells/interleukin-17 （Th17/IL-17） axis in the occurrence of vaginal eandidiasis in mice. Methods A total of 120 female BALB/c mice were randomly and equally divided into El, En, Ci and Cn groups. Three days before vaginal inoculation, estrogen （Ei and En） groups and control （Ci and Cn） groups received subcutaneous injection of 0.05 mg estradiol and 0.1 ml sterilized soybean oil at the hind legs, respectively, and then the hormone treatment continued every other day until the end of experiment. Infected （Ei and Ci） groups and noninfected （En and Cn） groups were inoculated intravaginally with 10 μ1 （5 × 10^4 conidia） of Candida albicans suspension and 10μ1of sterilized phosphate-buffered saline, respectively. Ten mice were randomly selected from each group and sacrificed on day 3, 7 and 14 after inoculation. The intact vagina tissues were resected and then frozen in liquid nitrogen or embedded in paraffin. Real-time fluorescence-based quantitative PCR （qRT-PCR） and immunofluorescent staining were performed to measure mRNA expression and immunofluorescence intensities of retinoic acid-related orphan receptorγt （RORγt）, RORα and IL-17, respectively. Western blot analysis was conducted to determine protein expression of RORγt and IL-17. Results Laser scanning confocal microscopy showed that RORγt, RORα and IL-17 immunofluorescence was mainly located at inflammatory cells of the lamina propria and blood vessels in En and Cn groups, at mucosal epithelium, adherent hyphae, and inflammatory cells of the lamina propria and blood vessels in Ci group, and at mucosal epithelium, vaginal canal and endocytosed hyphae, and inflammatory cells of the lamina propria and blood vessels in Ei group, qRT- PCR and immunofluorescent staining uncovered that mRNA expression and immunofluorescence intensities of RORγt, RORα and IL- 17 were significantly higher in En, Ci and Ei groups than in Cn group at the same time points （all P 〈 0.05）, as w
出处
《中华皮肤科杂志》
CAS
CSCD
北大核心
2017年第1期33-38,共6页
Chinese Journal of Dermatology
基金
国家自然科学基金（81171512）志谢广东医科大学统计学教研室倪少凯副教授进行统计学指导
