Experimental study for gene induction therapy to temporomandibular arthritis

颞下颌关节炎基因诱导治疗的实验研究

• 批准号: 13672112
• 负责人: TOMINAGA Kazuhiro
• 金额: $ 2.18万
• 依托单位: KYUSHU DENTAL COLLEGE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672112/
• 关键词: temporomandibular joint; arthritis; gene induction; electroporation; interleukin 1; animal; エレクトロポレーション; インターロイキン1

We constructed a plasmid vector of human interleukin 1 receptor antagonist (hIL-1ra) gene from U937 cell and confirmed the expression in in-vitro and in-vivo studies. The constructed gene was used for gene induction into the normal knee and the temporomandibular joints using with in-vivo gene electroporation technique. The production of hIL-1ra from those joints was confirmed with Western blotting but the production was too weak to reduce the arthritic condition.Therefore we tried to introduce the gene into peri-joint tissues including muscle tissue around the joint to enhance the anti-inflammatory protein production in adjuvant arthritis of the rat.1. Electroporatic gene induction and expression of hIL-1ra to a muscle : Various injected concentrations of DNA solution including the constructed vector were tested into the femoral muscle around the knee joint. The highest expression of the hIL-1ra was observed in the 50 micrograms' plasmid injection group.2. Duration of the gene expression : The expression of hIL-1ra was observed from one day to 28 days after electroporatic gene induction (50 micrograms' group).Conclusion : From our studies, electroporatic IL-1ra gene induction to the peri-joint tissues will be a possible option of the management of arthritic conditions because peri-joint muscle has a bioreactor effect to produce anti-inflammatory protein.

我们从U937细胞中构建了人白细胞介素1受体拮抗剂（hIL-1ra）基因的质粒载体，并在体外和体内研究中证实了其表达。利用体内基因电穿孔技术将构建的基因用于正常膝关节和颞下颌关节的基因诱导。 Western blotting证实这些关节产生hIL-1ra，但其产生太弱，不足以减轻关节炎状况。因此，我们尝试将该基因导入关节周围组织，包括关节周围的肌肉组织，以增强大鼠佐剂性关节炎中抗炎蛋白的产生。 1．电穿孔基因诱导和hIL-1ra在肌肉中的表达：将包括构建的载体在内的不同注射浓度的DNA溶液测试到膝关节周围的股骨肌肉中。 50μg质粒注射组hIL-1ra表达量最高。 2．基因表达持续时间：电穿孔基因诱导后1天至28天观察到hIL-1ra的表达（50微克组）。结论：根据我们的研究，对关节周围组织进行电穿孔IL-1ra基因诱导将是治疗关节炎病症的一种可能选择，因为关节周围肌肉具有产生抗炎蛋白的生物反应器效应。

项目成果

Kazuhiro Tominaga: "Interleukin-1beta in antigen-induced arthritis of the rabbit temporomandubular joint"Archives of Oral Biology. 46. 539-544 (2001)

Kazuhiro Tominaga：“抗原诱导的兔颞下颌关节关节炎中的白细胞介素 1β”口腔生物学档案。

Kazuhiro Tominaga: "Interleukin-1β in antigen-induced arthritis of the rabbit temporomandibular joint"Archives of Oral Biology. 46. 539-544 (2001)

Kazuhiro Tominaga：“抗原诱导的兔颞下颌关节关节炎中的白细胞介素 1β”口腔生物学档案 46. 539-544 (2001)。

Manabu Habu: "Immunohistochemical study of interleukin-1β and interleukin-1 receptor antagonist in an antigen-induced arthritis of the rabbit temporomandibular joint"Journal of Oral Pathology and Medicine. 31. 45-54 (2002)

Manabu Habu：“白细胞介素 1β 和白细胞介素 1 受体拮抗剂在抗原诱导的兔颞下颌关节关节炎中的免疫组织化学研究”口腔病理学与医学杂志 31. 45-54 (2002)。

Manabu Habu: "Immunohistochemical study of interleukin-1 beta and interleukin-1 receptor antagonist in an antigen induced arthritis of the rabbit temporomandibular joint"Journal of Oral Pathology and Medicine. 31. 45-54 (2002)

Manabu Habu：“白细胞介素 1 β 和白细胞介素 1 受体拮抗剂在抗原诱导的兔颞下颌关节关节炎中的免疫组织化学研究”口腔病理学与医学杂志。

M.Habu, K.Tominaga et al.: "Immunohistochemical study of interleukin-1Β and interleukin-1 receptor antagonist in an antigen-induced arthritis of the rabbit temporomandibular joint"Journal of Oval Pathology and Medicine. 31. 45-54 (2002)

M.Habu、K.Tominaga 等人：“抗原诱导的兔颞下颌关节关节炎中白细胞介素 1B 和白细胞介素 1 受体拮抗剂的免疫组织化学研究”《椭圆形病理学与医学杂志》31. 45-54 (2002)。 ）