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通过免疫组织化学和逆转录聚合酶链反应鉴定子宫内膜异位症中的巨噬细胞集落刺激因子受体

Identification of the M-CSF receptor in endometriosis by immunohistochemistry and RT-PCR.

作者信息

Mettler Liselotte, Schmutzler Andreas G, Koch Kerstin, Schollmeyer Thoralf, Salmassi Ali

机构信息

Department of Obstetrics and Gynecology, University Hospitals Schleswig-Holstein, Kiel, Germany.

出版信息

Am J Reprod Immunol. 2004 Nov;52(5):298-305. doi: 10.1111/j.1600-0897.2004.00228.x.

Abstract

PROBLEM

The aim of this paper is to provide further evidence that the dystopic proliferation of endometriotic epithelia is caused by the stimulation of peritoneal macrophages. It is essential to show that endometriotic epithelial cells express the macrophage colony-stimulating factor receptor (M-CSFR) which binds the M-CSF produced by the peritoneal macrophages.

METHOD OF STUDY

For the detection of M-CSFR, samples of ectopic endometrium (n = 79) and eutopic endometrium (n = 18) were compared. The specimens were gained at operative laparoscopy in the proliferative phase of the cycle. Cryostat sections were used for immunohistochemical detection. For in vitro reverse transcriptase polymerase chain reaction (RT-PCR) tests, the tissue was immediately shock frozen on paraffin sections. For the in situ RT-PCR technique the specimens were placed in a para-formaldehyde solution, embedded in paraffin and later processed. The Gene Amp 1000 in situ PCR system (Perkin Elmer) was used as the thermal cycler.

RESULTS

M-CSF and the M-CSF receptor are present in eutopic and ectopic endometrium. Qualitatively, with both PCR techniques we found the M-CSF receptor to be present in all samples examined. Using the histochemical detection technique, the M-CSF receptor was found in nearly 70% of endometriosis patients compared with a statistically significant lower percentage in normal endometrium.

CONCLUSIONS

The in situ RT-PCR technique and immunohistochemistry elaborated the need to trace the cellular sources of the M-CSF receptor. The identification of the M-CSF receptor in endometriotic tissue and in endometrium is apt to open a new experimental field in endometriosis research.

摘要

问题

本文旨在提供进一步证据,证明子宫内膜异位上皮的异位增殖是由腹膜巨噬细胞的刺激引起的。必须证明子宫内膜异位上皮细胞表达巨噬细胞集落刺激因子受体(M-CSFR),该受体可结合腹膜巨噬细胞产生的M-CSF。

研究方法

为检测M-CSFR,比较了异位子宫内膜样本(n = 79)和在位子宫内膜样本(n = 18)。样本在月经周期的增殖期通过手术腹腔镜检查获取。冰冻切片用于免疫组织化学检测。对于体外逆转录聚合酶链反应(RT-PCR)测试,将组织立即在石蜡切片上速冻。对于原位RT-PCR技术,将标本置于多聚甲醛溶液中,嵌入石蜡,随后进行处理。使用Gene Amp 1000原位PCR系统(Perkin Elmer)作为热循环仪。

结果

M-CSF和M-CSF受体存在于在位和异位子宫内膜中。定性地,通过两种PCR技术,我们发现M-CSF受体存在于所有检测样本中。使用组织化学检测技术,在近70%的子宫内膜异位症患者中发现了M-CSF受体,而在正常子宫内膜中的比例在统计学上显著较低。

结论

原位RT-PCR技术和免疫组织化学阐述了追踪M-CSF受体细胞来源必要性。在子宫内膜异位组织和子宫内膜中鉴定出M-CSF受体,有望在子宫内膜异位症研究中开辟一个新的实验领域。

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