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目的 :探索蛋白涂层支架携带质粒介导人肝脏诱导型一氧化氮合酶 (i NOS)基因转染小型猪冠状动脉可行性。 方法 :使用蛋白支架吸附去内毒素纯化质粒 ,以常规支架置入技术置入小型猪冠状动脉前降支中段。置入后第 7天取出前降支置入段 ,分别提取总核糖核酸 (RAN)并进行逆向多聚酶链反应 (RT- PCR) ,免疫组化染色检测导入人肝脏i NOS蛋白的表达。 结果 :小型猪前降支置入支架处显示人 i NOS基因信使核糖核酸 (m RNA)转录 ,免疫组化染色显示中膜、内膜人i NOS基因表达人 i NOS蛋白的颗粒 ,以平滑肌细胞最明显。 结论 :蛋白涂层支架吸附去内毒素携带人 i NOS基因质粒植入小型猪前降支冠状动脉 ,RT- PCR显示人 i NOS基因的 m RNA转录 ,免疫组化显示人 i NOS蛋白的表达。
OBJECTIVE: To explore the feasibility of protein-coated scaffolds carrying plasmid-mediated iNOS gene transfection into coronary arteries of miniature pigs. Methods: Purified plasmids of endotoxin were adsorbed by protein scaffolds and inserted into the middle part of anterior descending coronary artery of miniature pigs by conventional stent implantation. Total RNA was extracted and reverse-polymerase chain reaction (RT-PCR) was performed on day 7 after implantation. The expression of iNOS protein in human liver was detected by immunohistochemistry. Results: The mini-pigs were placed in the scaffold at the anterior descending branch of the pig to display the transcription of human iNOS gene messenger RNA (RNAi). Immunohistochemical staining showed that the iNOS gene in the media and intima expressed human iNOS protein, The most obvious. CONCLUSION: Protein-coated scaffolds are loaded with endotoxin to carry human iNOS gene plasmid implanted into the anterior descending coronary artery in mini-pigs. The m RNA transcripts of human iNOS gene are detected by RT-PCR and the expression of human iNOS protein is detected by immunohistochemistry.