Studies on physiologically active substances produced by plerocercoids of Spirometra erinaceieuropaei.

刺猬螺旋体产生的生理活性物质的研究。

• 批准号: 09670258
• 负责人: HIRAI Kazumitsu
• 金额: $ 2.05万
• 依托单位: Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670258/
• 关键词: Tape worm; Spirometra erinaceieuropaei; Growth hormone; Macrophage; Cytokine; Chemokine; TNF-α; iNOS; 腹腔マクロファージ; TNF-α抑制因子; プロテアーゼ; トリプシン・インヒビター; cDNA; 融合蛋白

1.The rule of a growth hormone-like factor and molecular cloning of the gene encoding it's factor produced by plerocercoids of Spirometra Erinaceieuropaei.A 1085 bp cDNA encoding a growth hormone-like factor (PGF) was cloned from a plerocercoid cDNA library. The amino acid sequence predicted from the cDNA has 65% homology with that of mouse cathepsin-L. Its fusion protein specifically reacted with anti-PGF antibody, but this fusion protein brought from E.coli XL-1 blue was not solublized with the buffer. The gene was characterized by Northern and Southern blot analysis, showing that PGF is stage specifically expressed. On the other hand, PGF had trypsin inhibitory activity, when the trypsin inhibitor was purified from the plerocercoid extract.2.Effects of excretory/secretory (ES) products of plerocercoids on murine peritoneal macrophages.ES products from plerocercoids reduced iNOS, chemokine and TNF-α mRNA levels in murine peritoneal macrophages stimulated with cytokines and/or LPS, analyzing with Northern blot. These suppressive factors were found in void fractions when ES products were separated by gel filtration. Then, macrophages obtained from plerocercoid-infected mice showed lower expression of TNF-α mRNA levels than those of uninfected controls. The suppressive effect of ES products was not depend on the prostaglandin E2 and IL-10.

1.一种生长激素样因子的产生规律及编码该因子基因的分子克隆。从plerocercoid cDNA文库中克隆了一个编码生长激素样因子（growth hormone-样factor， PGF）的1085 bp cDNA。预测的氨基酸序列与小鼠组织蛋白酶- l的同源性为65%。其融合蛋白与抗pgf抗体特异性反应，但该融合蛋白来自大肠杆菌XL-1蓝，不能与缓冲液溶解。Northern和Southern blot分析表明该基因是PGF分期特异性表达的。另一方面，PGF具有胰蛋白酶抑制活性，当胰蛋白酶抑制剂从蛤蚌提取物中纯化时。尾尾鱼的排泄/分泌（ES）产物对小鼠腹腔巨噬细胞的影响。在细胞因子和/或LPS刺激的小鼠腹膜巨噬细胞中，来自plerocercoids的ES产物降低了iNOS、趋化因子和TNF-α mRNA的水平。凝胶过滤分离ES产品时，在空隙馏分中发现了这些抑制因子。然后，从尾球蚴感染小鼠中获得的巨噬细胞中TNF-α mRNA的表达水平低于未感染的对照组。ES产物的抑制作用不依赖于前列腺素E2和IL-10。

项目成果

平井和光: "マンソン裂頭条虫擬充尾虫が産生する成長ホルモン様因子の宿主-寄生体関係における意義"米子医学雑誌. 50(3-4). 131-142 (1999)

Kazumitsu Hirai：“曼氏绦虫产生的生长激素样因子在宿主-寄生虫关系中的意义”米子医学杂志 50(3-4) (1999)。

H.Wang: "Excretory/secretory products of plerocercoids of Spirometra erinaceieuropaei induce the expression of inducible nitric oxide synthase mRNA in murine hepatocytes"Int. J. Parasitol.. 27. 367-375 (1997)

H.Wang：“Spirometra erinaceieuropaei 的 plerocercoids 的排泄/分泌产物诱导小鼠肝细胞中诱导型一氧化氮合酶 mRNA 的表达”Int。

H. Wang: "Excretory/secretory products from plerocercoids of Spirometra erinaceieuropaei induce the expression of inducible nitric oxide synthase mRNA in murine hepatocytes"International Journal for Parasitology. 27. 367-375 (1997)

H. Wang：“来自 Spirometra erinaceieuropaei 的 plerocercoids 的排泄/分泌产物诱导小鼠肝细胞中诱导型一氧化氮合酶 mRNA 的表达”国际寄生虫学杂志。

S. Fukumoto: "Excretory/secretory products from plerocercoids of Spirometra erinaceieuropaei reduce iNOS and chemokine mRNA levels in peritoneal macrophages stimulated with cytokine and or LPS"Parasite Immunology. 19. 325-332 (1997)

S. Fukumoto：“来自 Spirometra erinaceieuropaei 的 plerocercoids 的排泄/分泌产物降低了用细胞因子和/或 LPS 刺激的腹膜巨噬细胞中的 iNOS 和趋化因子 mRNA 水平”寄生虫免疫学。

H.Wang: "Molecular cloring and expression of the gene encording a growth factor of Spirometra erinaceieuropaei." 9^<th> International Congress of Parasitology. 831-834 (1998)

H.Wang：“编码螺旋藻生长因子的基因的分子克隆和表达。”