米非司酮对子宫内膜异位症大鼠内膜OPN及MMP-9表达的影响

摘要

目的 探讨米非司酮对大鼠子宫内膜异位症(Endometriosis,EMS)模型在位及异位内膜中骨桥蛋白(OPN)及金属蛋白酶9(MMP-9)表达的影响.方法 取建模成功的13只大鼠随机分为米非司酮组(7只),对照组(6只),药物干预4周后开腹取材,通过免疫组织化学法及RT-PCR法测定药物干预前、后在位及异位内膜中MMP-9及OPN的表达.结果 干预后米非司酮组在位内膜和异位内膜中OPN及MMP-9的表达均较干预前表达明显降低(P＜ 0.05);对照组干预后异位内膜中OPN的表达强于在位内膜,而米非司酮组干预后异位内膜及在位内膜中OPN表达差异无统计学意义(P=0.113);对照组干预后异位内膜中MMP-9的表达强于在位内膜,且干预后米非司酮组异位内膜中MMP-9的表达也较在位内膜中表达增强(P=0.011);对照组干预后异位内膜中OPNmRNA表达较干预前明显减低,且在位内膜及异位内膜的表达差异无统计学意义.结论 米非司酮可降低大鼠EMS模型在位、异位内膜中OPN及MMP-9的表达,对异位内膜中OPN的抑制作用要强于在位内膜;对在位内膜中MMP-9的抑制作用不弱于对异位内膜.米非司酮可能通过抑制病灶局部OPN、MMP-9的表达而降低细胞的黏附和侵袭能力.%Objective To construct the recombinant plasmid pET -pip and to detect its expression in the pro-karyotic cells BL2l( DE3 ). Methods The pip gene was amplified by PCR from DNA of Legionella pneumo-phila, then inserted it into the prokaryotic expression vector pET -32a( + ). The recombinant plasmid pET -pip was constructed. The recombinant plasmid was identified by restriction - endonuclease digestion, PCR and DNA sequencing analysis and transferred into BL21. The expression of pET -pip was induced with IPTG and the fusion protein was analyzed with SDS - PAGE. It was purified by His - tag. Results The pip gene of 726bp in length was amplified and the recombinant plasmid pET -pip was constructed. The PIP protein of approximately 46KD in size was expressed in E. Coli and purified. Conclusion The pip gene of 726bp in length was successfully cloned and the recombinant plasmid pET -pip was constructed successfully and expressed efficiently.