The development of new strategy of heart preservation with. transfection of a dominant-negative inhibitor of monocyte chemoattractant protein-1 gene

心脏保存新策略的制定。

• 批准号: 14571271
• 负责人: MORITA Shigeki
• 金额: $ 2.37万
• 依托单位: Kyushu 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 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14571271/
• 关键词: renerfusion injury; MCP-1; 再還流障害; 遺伝子療法

Background : Monocyte chemoattractant protein-1 (MCP-1), a potent chemotactic factor for monocytes, is induced during ischemia-reperfusion. As monocytes might play an important causative role in reperfusion injury, we investigated if inhibition of monocyte activation could attenuate ischemia-reperfusion injury and thereby improve cardiac preservation. To inhibit monocyte activation, we transfected a dominant-negative inhibitor of MCP-1 (7ND) gene in an animal model. Methods and Results : We used an isolated rabbit heart preparation perfused with support-rabbit blood and transfected 7ND genes to skeletal muscle of the support rabbits (n=7) using electroporation technique ; causing an elevation of serum 7ND level to 20 ± 7pg/ml at 5 days after transfection. Animals receiving empty plasmid served as controls (n=7). Five days after, transfection, hearts from other rabbits were excised, stored in UW solution for 6hours, and perfused with blood from transfected support rabbits. The 7ND group showed better cardiac output (128.7 ± 17.9 vs. 81.6 ± 19.8ml/min: p<0.01), lower serum CK-MB levels (5.0 ± 1.8 vs. 11.1 ±2.9ng/ml:p<0.01), lower serum IL-1 β levels (257.2 ± 23.2 vs. 311.2 ± 37.4pg/ml: p<0.05) and lower serum TNF-αlevels (19.0 ± 8.4 vs. 35.1 ± 13.0pg/ml: p<0.05). The numbers of infiltrating cells in myocardium were significantly reduced in the 7ND group. Conclusions: Inhibition of MCP-1 with 7ND gene transfection reduced cytokine activation, attenuated myocardial damage, and improved cardiac function after 6-hours of preservation. These results show that MCP-1 plays an important role in ischemia-reperfusion injury.

背景：单核细胞趋化蛋白-1（MCP-1）是一种强有力的单核细胞趋化因子，在缺血再灌注过程中被诱导表达。由于单核细胞可能在再灌注损伤中起重要的致因作用，我们研究了抑制单核细胞活化是否可以减轻缺血-再灌注损伤，从而改善心脏保护。为了抑制单核细胞活化，我们在动物模型中转染了MCP-1的显性负性抑制剂（7 ND）基因。方法和结果：用离体兔心脏标本灌注支持体兔血，用电穿孔技术将7 ND基因转染到支持体兔（n=7）的骨骼肌中，转染后5天血清7 ND水平升高至20 ± 7 pg/ml。接受空质粒的动物作为对照（n=7）。转染后5天，切下其他兔的心脏，在UW溶液中保存6小时，并用转染的支持兔的血液灌注。7 ND组心输出量较好（128.7 ± 17.9 vs. 81.6 ± 19.8ml/min：p<0.01），血清CK-MB水平降低（5.0 ± 1.8 vs. 11.1 ±2.9ng/ml：p<0.01），降低血清IL-1 β水平（257.2 ± 23.2 vs. 311.2 ± 37.4pg/ml：p<0.05）和降低血清TNF-α水平（19.0 ± 8.4 vs. 35.1 ± 13.0pg/ml：p<0.05）。7 ND组心肌浸润细胞数明显减少。结论：用7 ND基因转染抑制MCP-1可减少细胞因子活化，减轻心肌损伤，并在保存6小时后改善心功能。这些结果表明MCP-1在缺血再灌注损伤中起重要作用。

项目成果

Kajihara N, Morita S, et al.: "Transfection with a Dominant-Negative Inhibitor of Monocyte Chemoattractant Protein-1 Gene Improves Cardiac Function after 6 hours of Cold Preservation"Circulation. 108,19. II213-II218 (2003)

Kajihara N、Morita S 等人：“转染单核细胞趋化蛋白 1 基因的显性负性抑制剂可改善冷保存 6 小时后的心脏功能”循环。

Kajihara N, Morita S, et al.: "Transfection with a Dominant-Negative Inhibitor of Monocyte Chemoattractant Protein-1 Gene Improves Cardiac Function after 6 hours of Cold Preservation"Circulation. 106,19. 549 (2002)

Kajihara N、Morita S 等人：“转染单核细胞趋化蛋白 1 基因的显性负性抑制剂可改善冷保存 6 小时后的心脏功能”循环。

Kajihara N, Morita S, et al.: "Transfection with a Dominant-Negative Inhibitor of Monocyte Chemoattractant Protein-1 Gene Improves Cardiac Function after 6 hours of Cold Preservation"Circulation. 108. II213-II218 (2003)

Kajihara N、Morita S 等人：“转染单核细胞趋化蛋白 1 基因的显性负性抑制剂可改善冷保存 6 小时后的心脏功能”循环。

Kajihara N, Morita S, et al.: "Transfection with a Dominant-Negative Inhibitor of Monocyte Chemoatractant Protein-1 Gene Improves Cardiac Function after 6 hours of Cold Preservation"Circulation. 108. II213-II218 (2003)

Kajihara N、Morita S 等人：“转染单核细胞趋化蛋白 1 基因的显性负性抑制剂可改善冷保存 6 小时后的心脏功能”循环。

Kajihara N, Morita S, et al.: "Transfection with a Dominant-Negative Inhibitor of Monocyte Chemoattractant Protein-1 Gene Improves Cardiac Function after 6 hours of Cold Preservation"Circulation. 106・19. Supple P549 (2002)

Kajihara N、Morita S 等：“转染单核细胞趋化蛋白 1 基因的显性阴性抑制剂可改善冷保存 6 小时后的心脏功能”循环 106・19。