人源性双亲性穿膜肽PKV14的合成及其对BHK-21细胞的影响.PDFVIP

·2482 · 现代药物与临床 Drugs Clinic 第 33 卷 第 10 期 2018 年 10 月 人源性双亲性穿膜肽 PKV14 的合成及其对BHK-21 细胞的影响 1 1 2* 陈 莹 ，宋金春 ，刘岩松 1. 武汉大学人民医院 药学部，湖北 武汉 430060 2. 武汉回盛生物科技股份有限公司，湖北 武汉 430000 摘 要：目的 合成人源性双亲性穿膜肽 RRTLRRRRAAQRCG （PKV14 ），并研究其细胞穿膜效果和对细胞活性的影响。 方法 采用固相合成法合成 PKV14 ，采用高效液相色谱和质谱对 PKV14 进行验证，使用荧光显微镜和酶标仪观察 PKV14 在 BHK-21 细胞系中的穿膜效果，采用 MTT 实验法检测 PKV14 的细胞毒性。结果 合成了 PKV14 肽段，并且 PKV14 对 于 BHK-21 细胞系的穿膜效率极高，细胞毒性实验表明 PKV14 对 BHK-21 细胞活性无影响。结论 PKV14 是具有高穿膜活 性，安全低毒的新型人源性细胞穿膜肽，有作为药物载体的潜力。 关键词：人源性双亲性穿膜肽；PKV14 ；穿膜活性；BHK-21 细胞 中图分类号：R914.2 ；R966 文献标志码：A 文章编号：1674 - 5515(2018)10 - 2482 - 05 DOI: 10.7501/j.issn.1674-5515.2018.10.003 Synthesis of human-derived amphipathic cell-penetrating peptide PKV14 and effect on BHK-21 cell 1 1 2 CHEN Ying , SONG Jin-chun , LIU Yan-song 1. Department of Pharmacy, Renmin Hospital of Wuhan University, Wuhan 430060, China 2. HVSEN Biotech Co., Ltd, Wuhan 430000, China Abstract: Objective To synthesize the human-derived amphipathic cell-penetrating peptide RRTLRRRRAAQRCG (PKV14), and investigate the effect on cell membrane penetration and its cytotoxicity. Methods PKV14 was synthesized by a solid-phase peptide synthesis. HPLC and MS analysis methods were applied to identifying its structure. The penetrating effects of PKV14 in BHK-21 cells were tested by fluorescence microscopy and microplate reader. The cytotoxicity of PKV14 was tested by MTT method. Results PKV14 was synthesized successfully and it was confirmed that PKV14 had a very high penetrating efficiency in BHK-21 cells. Cytotoxicity test showed that PKV14 had no effect on the activity of BHK-21 cells. Conclusion PKV14 is a novel human cell penetrating peptide with high penetrating activity, safety, a