白念珠菌对角质形成细胞β防御素-2 mRNA表达的影响

The Effect of Candida albicans on the Expression Level of Human β-defensin-2 mRNA in Keratinocytes in vitro

目的探讨白念珠菌及其菌体成分对人角质形成细胞β防御素-2表达的影响。方法采用酶解、反复冻融、超声破碎和超速离心等方法制备真菌成分;用逆转录聚合酶链反应(RT-PCR)方法检测白念珠菌及其菌体成分刺激人角质形成细胞24h后β防御素-2mRNA表达。结果正常培养的人角质形成细胞和角质形成细胞永生细胞系HaCaT均有β防御素-2mRNA的少量表达,白念珠菌、白念珠菌细胞壁提取物和纯甘露聚糖可以使β防御素-2mRNA表达增强(P值分别<0.01、0.05和0.01),而白念珠菌培养上清液和细胞浆提取物对β防御素-2表达无明显影响(P>0.05)。结论白念珠菌可上调角质形成细胞β防御素-2mRNA的表达,甘露聚糖可能是起诱导作用的活性成分。角质形成细胞通过表达β防御素-2在皮肤黏膜的抗感染防御机制中发挥重要作用。

Objective To investigate the effect of Candida albicans and its components on the expression level of human beta-defensin-2 mRNA (HBD-2) in keratinocytes in vitro. Methods Different components of Candida albicans were isolated by lyticase, repeated freezing and thawing, sonication, and centrifugation. The keratinocytes and HaCaT cell lines were co-cultured with Candida albicans and its cellular components for 24 h. The expression level of HBD-2 mRNA was detected by reverse-transcription polymerase chain reaction (RT-PCR). Results Low expression level of HBD-2 mRNA in the unstimulated keratinocytes and HaCaT cells was detected. The HBD-2 mRNA expression levels in the keratinocytes stimulated by Candida albicans, the extract of its cell wall, and pure mannan were significantly increased (P < 0.01, P < 0.05, and P < 0.01, respectively). No significant differences of the HBD-2 mRNA expression levels of the keratinocytes stimulated with the cultural supernatant and plasm extract of Candida albicans (P > 0.05). Conclusions Candida albicans, the extract of cell wall of Candida albicans, and commercial mannan can increase the expression level of HBD-2 mRNA in keratinocytes.