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INVOLVEMENT OF APOPTOTIC PATHWAY IN PATHOGENESIS OF SJOGRENS SYNDROME

凋亡途径参与干燥综合征的发病机制

基本信息

批准号：
14571935
负责人：
SAITO Keiichi
金额：
$ 2.43万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571935/
关键词：
SJOGREN'S SYNDROME APOPTOSIS MITOCHONDRIA TNF ALPHA IMMUNOHISTOCHEMISTRY MRL lprマウス

项目摘要

We examined immunohistochemically the involvement of TNF α-mediated and mitochondria-mediated apoptotic pathway in pathogenesis of Sjogren's syndrome using autoimmune model mouse, MRL/lpr mice, and control mice, MRL/+ mice, in our present study. 5-month-old mice of each strain were sacrificed by anesthesia overdose followed by exsanguinations. Submandibular glands were removed, fixed in 10% buffered formalin, and embedded in paraffin. Paraffin-embedded tissue sections (3μm) were immunostained against TNF α, TNF receptor 1, Fadd, caspase 8, caspase 3, which are implicated in TNF α-mediated apoptotic pathway, in addition to caspase 2, Bid, p53, PUMA Bax, cytochrome c, Apsf-1, caspase 9, which are related to mitochondria-mediated apoptosis, using streptoavidin-biotin method. The specimens of MRL/lpr mice showed intense expression of these ligands in ductal cells of submandibular glands. However, the immunolocalizations of these ligands in MRL/+ mice were weak and not convincing.It has been revealed that tissue destruction of salivary glands observed in Sjogren's syndrome is triggered by apoptosis, and Fas-Fas ligand apoptotic pathway plays a crucial role in the pathogenesis of this disease. We have also confirmed that, impaired submandibular gland tissues of MRI/lpr mice attribute to the commitment to apoptosis through ssDNA expression. Furthermore, Fas-Fas ligand apoptotic pathway is abrogated in MRL/lpr mice because they carry a mutant of fans gene. Therefore, our present, study indicates that TNF α-mediated and mitochondria-mediated apoptotic pathway other than Fas-Fas ligand apoptotic pathway has the association with the development, of Sjogren's syndrome-like sialadenitis in MRL/lpr mice.

在本研究中，我们利用自身免疫模型小鼠MRL/lpr小鼠和对照小鼠MRL/+小鼠，用免疫组织化学方法检测了TNF α介导和线粒体介导的凋亡通路在干燥综合征发病机制中的作用。各组5月龄小鼠均采用麻醉过量后放血处死。取下颌下腺，10%福尔马林缓冲固定，石蜡包埋。采用链亲素-生物素法对石蜡包埋组织切片（3μm）进行TNF α、TNF受体1、Fadd、caspase 8、caspase 3和与线粒体介导的凋亡相关的caspase 2、Bid、p53、PUMA Bax、细胞色素c、Apsf-1、caspase 9的免疫染色。这些配体在MRL/lpr小鼠颌下腺导管细胞中表达强烈。然而，这些配体在MRL/+小鼠中的免疫定位较弱且不令人信服。研究发现，干燥综合征涎腺组织破坏是由细胞凋亡引起的，Fas-Fas配体凋亡通路在干燥综合征的发病机制中起着至关重要的作用。我们也证实，MRI/lpr小鼠的颌下腺组织受损与ssDNA表达承诺凋亡有关。此外，由于MRL/lpr小鼠携带fan基因突变，Fas-Fas配体凋亡途径在MRL/lpr小鼠中被取消。因此，我们目前的研究表明，除了Fas-Fas配体凋亡途径外，TNF α介导的凋亡途径和线粒体介导的凋亡途径与MRL/lpr小鼠干燥综合征样涎腺炎的发生有关。