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目的探讨由胰蛋白酶原基因(cationic trypsinogen,PRSS1)突变引发的早发型自身免疫性相关的多器官多发囊肿及其致病机制。方法采用DNA全长测序技术分析PRSS1、囊性纤维化跨膜通道调节因子(cystic fibrosis transmembrane conductance regulator,CFTR)、丝氨酸蛋白酶抑制剂Kazal 1型(setine protease inhibitor Kazal type 1,SPINK1)、蛋白激酶D(protein kinase D,PKD)1和PKD2等胰腺炎和多囊性病变相关基因的所有外显子及其侧翼内含子剪切区域,确定DNA和cDNA序列的变异,通过与家系内部和正常对照的比较分析,对检测到的变异是否与疾病相关进行探讨,并构建突变体表达体系进行功能学验证,同时对患者的肺、肝、胰腺等穿刺样本进行免疫组织化学和特殊染色。结果在2例年轻的自身免疫性胰腺炎患者中首次发现PRSS1基因2号外显子缺失突变生成激活肽缺失型的胰蛋白酶原,并具有生物学活性;肝脏、肺穿刺病理均可见不同程度的淋巴细胞和浆细胞浸润,肺组织病理显示弹力纤维、网状纤维明显减少;患者表现为多脏器多囊性病变,血清胰蛋白酶、弹力蛋白酶、AAT显著增高。使用糖皮质激素治疗有效。结论 PRSS1:c.300_1304 del CCCAG是引发早发型自身免疫性胰腺炎的新突变形式,并与多器官囊肿关系密切。
Objective To investigate the early-onset autoimmune-associated multiple organ multiple cysts and its pathogenesis induced by the mutation of the human trypsinogen (PRSS1) gene. Methods DNA sequencing was used to analyze the expression of PRSS1, cystic fibrosis transmembrane conductance regulator (CFTR), serine protease inhibitor Kazal type 1 (SPINK1) and protein kinase D (PKD) 1 and PKD2 and other pancreatitis and polycystic lesions related genes all exons and their flanking intron cutting region, to determine the DNA and cDNA sequence variation, with the family and the normal control The differences between the detected variants and the disease were discussed, and the mutant expression system was constructed for functional verification. At the same time, immunohistochemistry and special staining were performed on the puncture samples such as lung, liver and pancreas. Results The deletion mutation of exon 2 of PRSS1 gene was found in 2 young patients with autoimmune pancreatitis for the first time to produce tryptase with active peptide deletion type and had biological activity. Pathological changes of liver and lung were all observed in different degree of lymph nodes Cells and plasma cells infiltration, pathological lung showed elastic fibers, reticular fibers was significantly reduced; patients showed multiple organ polycystic lesions, serum trypsin, elastase, AAT was significantly higher. Glucocorticoid treatment is effective. Conclusion PRSS1: c.300_1304 del CCCAG is a new mutation that causes early-onset autoimmune pancreatitis and is closely related to multiple organ cysts.