阴道菌群改变与宫颈癌前病变相关性的初步研究

The correlation between changes in vaginal flora and cervical lesions

目的本实验对130例健康、HPV感染[HPV(+)]及不同级别宫颈病变(CIN)患者阴道菌群进行研究分析,揭示阴道菌群变化与宫颈病变的发生及HPV感染的关系,为阻断宫颈病变的进一步发展提供理论依据。方法收集2014-2015年大连市妇幼保健院门诊就诊的20~65岁患者130例,依据宫颈HPV检测和阴道镜下宫颈活组织检查病理诊断结果进行分组。采用QIAamp DNA mini和Blood mini试剂盒提取阴道菌群基因组DNA。利用PCR-DGGE技术对患者阴道菌群的变化进行分析,Quantity One软件进行聚类分析及凝胶测序法分析特异条带的种类。上述数据的处理和分析采用SPSS 19.0统计软件。结果在DGGE图谱中,HPV(+)组阴道菌群丰度高于HPV(-)组。CIN组阴道菌群丰度较HPV(+)组升高。测序结果显示,HPV(-)组与HPV(+)组中3条共有强度较大的条带均属于厚壁菌门,HPV(+)组的特征谱带为阴道加德纳菌(Gardnerella vaginalis)。聚类分析和主成分分析(PCA)显示,HPV(+)组患者发生了菌群失调。结论乳酸菌为阴道菌群中的优势菌;放线菌是HPV(+)的CIN患者的特殊细菌门,该菌属于阴道加德纳菌;HPV感染可引起阴道菌群稳态失调,且宫颈病变程度越重,阴道菌群失衡情况也相应加重。

Objective To reveal the correlation between the changes in vaginal flora and the development of cervical intraepithelial neoplasia（CIN）and HPV infection,and provide theoretical support for the upcoming studies that focus on blocking the development of CIN.Methods We randomly enrolled 130 patients aged 20 to 65 years from the gynecological clinic of Dalian Maternal and Child Health Hospital from 2014 to2015.The specimens were further divided into 4 groups based on the results of HPV detection and colposcopic cervical biopsy.QIAamp Mini DNA and Blood Mini Kits were used to extract DNA pairs from vaginal flora.PCR-DGGE was used to analyse the changes of vaginal flora.Different types of specific stripes were identified with cluster analysis and rubber cutting sequencing.All the data were processed with SPSS 19.0 Data Analyzing software.Results DGGE graph indicated that the abundance of vaginal flora in HPV（＋）group was higher than in HPV（-）group,and the abundance of vaginal flora in the CIN group was higher compared with the HPV（＋）group.The result of sequencing test showed that the three stronger strips that appeared in both HPV（-）and HPV（＋）groups belonged to Phylum Firmicutes,and the characteristic band of the HPV（＋）group was Gardnerella vaginalis.Besides,the results of cluster analysis and PCA revealed that flora imbalance occurred in patients in the HPV（＋）group.Conclusion Lactic acid bacteria are predominant among vaginal flora.Actinomycete is the special bacterial phylum of CIN patients with positive HPV,which belongs to the category of Gardnerella vaginalis.HPV infection can cause flora imbalance which is positively associated with the severity of cervical lesions.