阴道上皮细胞先天性抗假丝酵母菌的作用研究

Congenital anti-candida of human vaginal epithelial cells

目的:以人阴道上皮细胞和人阴道灌洗液为模型,分析与阴道上皮细胞先天性抗假丝酵母菌机制有关的细胞因子在假丝酵母菌感染中的变化,研究阴道上皮对抗外阴阴道假丝酵母菌病(vulvovaginal candidiasis,VVC)的作用。方法:用组织块法和含表皮生长因子的无血清培养基体外培养人阴道上皮原代细胞,实验组(VVC组)为细胞传代后与培养白假丝酵母菌的上清液共同培养,对照组为传代细胞加无血清培养基,在0、3、6、12、24、48h收集各组的细胞上清液,用酶联免疫吸附测定法(ELISA)测定阴道上皮细胞分泌的人β防御素-1(human β defensin-1,HBD-1)、人β防御素-2(human β defensin-1,HBD-2)、表面活性蛋白A(surfactant protein-A,SP-A)等细胞因子的变化,并取VVC患者、复发性VVC(RVVC)患者及健康者的阴道灌洗液用ELISA方法分别测定SP-A变化,对实验数据进行统计学分析。结果:在培养的人阴道上皮细胞模型组中,VVC组的HBD-1(F=62.784,P=0.001)、HBD-2(F=5127.984,P=0.000)和SP-A(F=542.210,P=0.000)的表达均明显高于对照组。在人阴道灌洗液模型组中,VVC组的SP-A明显高于对照组(P=0.021),RVVC组的SP-A高于对照组及VVC组。结论:人阴道上皮细胞有先天性抗白假丝酵母菌作用,能分泌HBD-1、HBD-2、SP-A。白假丝酵母菌感染人阴道上皮细胞时,HBD-1、HBD-2、SP-A的分泌量均有增加。RVVC组的SP-A水平可能比VVC组更高。

Objective： To further investigate pathogenesis about vulvovaginal candidiasis （VVC）. Methods： Human normal vaginal epithelial cells in vitro were cultured by tissue culture and keratinocyto serum-free medium. Passaging epithelial cells were cocultured with candida albicans medium in separate wells for 0, 3, 6, 12, 24, and 48 h. Control epithelial cells were cultured alone in separate wells. For examination of cytokines and chemokines, at each time point, the coculture supernatant and the control culture were collected. ELISA was applied to detect the levels of HBD-1, HBD-2 and SP-A protein expression in the vaginal epithelial cells （ HBD-1 ：F = 62.784,P = 0. 001 ; HBD-2： F = 5127. 984, P = 0.000 ; F = 542.210, P = 0.000）. Also, we collected the vaginal doughing water of VVC patients and healthy women, and detected the levels of SP-A, then analysed the data. Results： In the trail of human vaginal epithelial cells, in contrast to the control group, the medium group induced the epithelial cells to secrete more HBD-1, HBD-2 and SP-A at h48, which was considered to be significant （ P 〈 0.05 ）. In the trail of human vaginal doughing water, in contrast to the control group, the levels of SP-A of VVC group were higher, which was considered to be significant （P 〈 0.05 ）. The levels of SP-A of rucurrent vulvovaginal candidiasis （RVVC） group were higher than those of VVC group and control group. Conclusion： Human vaginal epithelial cells function as congenital anti-Candida albicans and can secrete HBD-1, HBD-2 and SP-A. When infected by Candida albicans, human vaginal epithelial cells can secrete HBD-1, HBD-2 and SP-A The levels of SP-A of RVVC patients are probably higher than those of VVC patients.