人宫颈黏液HMGN2鉴定及在宫颈组织的表达

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为探讨人子宫颈黏液的抗菌机制,采用酸性尿素聚丙烯酰胺凝胶电泳和反相高压液相色谱技术,从人子宫颈黏液酸溶性提取物鉴定出对大肠杆菌氨苄青霉素耐药株ML-35p有较强的抗菌活性的多肽HCP-21和HCP-26.蛋白质N端氨基酸序列测定和精确质谱分子质量测定证明,HCP-21为HMGN2,HCP-26为SLPI片段.提取原代培养宫颈上皮细胞的总RNA,采用RT-PCR技术扩增出一条与HMGN2 cDNA大小相同(270bp)目的基因片段,表明在生理状况下,宫颈上皮细胞即可表达其mRNA.制备HMGN2多克隆抗体,对生理状态下宫颈组织切片和宫颈黏液涂片进行HMGN2分子免疫组化分析表明,该分子主要分布于子宫颈黏膜层,并存在于子宫颈黏液.HMGN2分子在宫颈黏膜上皮组织和黏液中固有表达,可能在子宫颈天然免疫防御中起重要作用. In order to explore the antibacterial mechanism of human cervical mucus, acidic urea polyacrylamide gel electrophoresis and reverse phase high pressure liquid chromatography were used to detect the inhibitory effect of the ampicillin-resistant E. coli strain ML-35p HCP-21 and HCP-26, which have strong antibacterial activity, were determined by N-terminal amino acid sequence analysis and precise mass spectrometry molecular mass spectrometry.HCP-21 was identified as HMGN2 and HCP-26 as SLPI fragment.The primary culture of cervical epithelial cells Of total RNA was amplified by RT-PCR and a HMGN2 cDNA of the same size (270bp) gene fragment, indicating that under physiological conditions, the cervical epithelial cells can express their mRNA.Preparation of HMGN2 polyclonal antibody to physiological state Cervical tissue sections and cervical mucus smears HMGN2 molecular immunohistochemical analysis showed that the molecule is mainly distributed in the cervical mucosa and exists in cervical mucus.HMGN2 molecules in cervical mucosa epithelial tissue and mucus inherent expression may be in the cervix Natural immune defense plays an important role.
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