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Research on the pathogenic mechanism of pulmonary hypertension (PH) in mixed connective tissue disease (MCTD).

混合性结缔组织病（MCTD）肺动脉高压（PH）发病机制研究。

基本信息

批准号：
15590312
负责人：
SAWAI Takashi
金额：
$ 1.86万
依托单位：
Iwate Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590312/
关键词：
mixed connective tissue disease (MCTD)pulmonary hypertension anti-endothelial cell antibody (AECA)アポトーシス NK細胞サイトカイン IL-2 血管内皮細胞 MCTD AECA Fas

项目摘要

The pathogenesis of pulmonary hypertension (PH) in mixed connective tissue disease (MCTD) is still unclear. Recent studies have suggested that PH in MCTD is resulted from endothelial cell injury. To reveal the pathogenic mechanism of PH in patients with MCTD, this research was focused on anti-endothelial cell antibodies (AECA) and identification of target proteins of AECA.Cyto-ELISA was performed to detect AECA in MCTD patients with PH using micro vascular endothelial cell (HMVEC-L). To determine AECA prevalence, values higher than 2 S.D. above the mean of healthy control were considered as positive. Six of 9 MCTD patients had positive results. The titer of AECA was higher in MCTD patients with PH than without PH.For identification of targets of AECA, proteins from HMVEC-L were separated by two-dimensional electrophoresis and transferred on PVDF membranes. Western blot analyses were performed with sera from healthy control or MCTD patients. Detected protein spots with a difference among the samples were selected and identified by peptide mass finger printing. About 20 protein spots were detected and half of these were identified.As the results of two-dimensional western blot analyses, candidate antigens were obtained. These included proteins reported as autoantigen in various diseases. Furthermore, one of candidates is suggested to be associated with PH in patients with SSc. This raises possibility that the identified proteins include antigens of AECA, indicating that two-dimensional western blot analysis is useful technique for detection of the target antigens of AECA.

混合性结缔组织病（MCTD）中肺动脉高压（PH）的发病机制尚不清楚。最近的研究表明MCTD中的PH是由内皮细胞损伤引起的。为揭示MCTD患者PH的发病机制，本研究重点开展抗内皮细胞抗体（AECA）及其靶蛋白的鉴定。利用Cyto-ELISA技术，利用微血管内皮细胞（HMVEC-L）检测MCTD合并PH患者的AECA。为了确定 AECA 患病率，值高于 2 S.D.高于健康对照的平均值被认为是阳性。 9 名 MCTD 患者中有 6 名结果呈阳性。患有PH的MCTD患者的AECA滴度高于未患有PH的MCTD患者。为了鉴定AECA的靶标，通过二维电泳分离来自HMVEC-L的蛋白质并转移到PVDF膜上。使用健康对照或 MCTD 患者的血清进行蛋白质印迹分析。选择样品中检测到的具有差异的蛋白质点并通过肽质量指纹图谱进行鉴定。检测到约20个蛋白点，其中一半得到鉴定。二维Western blot分析结果，获得候选抗原。其中包括据报道在各种疾病中作为自身抗原的蛋白质。此外，其中一种候选药物被认为与 SSc 患者的 PH 相关。这提出了所鉴定的蛋白质包括AECA抗原的可能性，表明二维蛋白质印迹分析是检测AECA靶抗原的有用技术。