目的 检测阴道念珠菌病患者体内感染状态下白念珠菌抗氧化基因的表达,初步了解抗氧化机制在阴道念珠菌病发病机制中的关系.方法 选取7例阴道念珠菌病患者阴道分泌物分离株,培养鉴定为白念珠菌,选取PDA斜面培养3天(体外培养静止期)及临床阴道分泌物检体,用Trizol法提取总RNA反转录为cDNA后进行荧光定量PCR,检测体外培养静止期与体内检体抗氧化基因(CAT,SOD2,SOD5)及抗氧化转录因子(CaSkn7,CaHog1,CAP1)的基因表达情况,运用Ct值比较法进行表达量的相时定量分析.结果 白念珠菌阴道病患者体内标本中CaSkn7,CAP1,SOD2,SOD5基因表达量比体外培养静止期少,分别为体外表达量的0.273,0.635,0.039,0.632;而CaHog1及CAT基因表达比体外培养静止期高,分别为体外表达量的3.5倍及1.67倍.经Wilcoxon检验,显示阴道分泌物标本中仅CaSkn7基因表达量较体外培养静止期低,差异有统计学意义P〈O.05,其余基因与体外培养静止期差异无统计学意义,P均〉0.05.结论 白念珠菌黏膜感染状态下抗氧化相关的基因表达较少,抗氧化机制在念珠菌性阴道炎发病过程中作用不明显.
Objective To evaluate the anti-oxidative relative gene expression in samples from vaginal candidiasis, and assay the anti-oxidative stress response in vivo vaginal candidasis. Methods Seven samples from vaginal candidiasis were cultured in PDA for 3 days ( in vitro stationary phase). The total RNA of C. albicans in vitro cultures and in vitro samples were isolated by Trizol method , and then performed reverse transcription real-time PCR. The comparative Ct method were used to analyze relative genes expression of anti-oxidant en- zyme (CAT,SOD2,SODS) and anti-oxidative related transcription factors (CaHogl, CaSkn7, CAP1 ) of C. albicans in vitro and in vivo. Results In vivo vaginal candidiasis samples, the expression of CaSkn7, CAP1, SOD2, SOD5 were at low level compared with in vitro stationary phase with 0. 273, 0.635, 0.039, 0.632-times and the expression of CaHogl and CAT were 3.5 and 1.67-times higher than in vitro stationary phase. The low expression level of CaSkn7 had statistically significant (wilcoxon signed-rank matched-pair test P 〈 0.05 ), while others genes'expression showed no statistically significant. Conclusion The low ex- pression level of anti-oxidant related gene in vivo vaginal candidasis suggest that little oxidative stress response in pathogenesis of vaginal candidasis.