C型利钠肽对血管内皮细胞株增殖能力的影响

Influence of human C-type natriuretic peptide on vascular endothelial cell proliferation

目的了解C型利钠肽(CNP)对血管内皮细胞增殖能力的影响。方法将所构建的含人CNP(hCNP)的重组质粒pcDNA3.1(+)在聚乙烯亚胺介导下转染人脐静脉血管内皮细胞株。通过反转录-聚合酶链反应、免疫组织化学和蛋白质印迹法检测该质粒的表达,用噻唑蓝法检测该质粒的表达产物对人脐静脉血管内皮细胞株增殖能力的影响。同法转染下列物质作对照:pcDNA3.1 (+)(阴性对照)、增强型绿色荧光蛋白质粒(阳性对照,只用于检测转染率)、磷酸盐缓冲液(空白对照)。结果pcDNA3.1(+)转染48 h后人脐静脉血管内皮细胞株的增殖能力为0.164±0.012;与其比较,含hCNP的pcDNA3.1(+)在血管内皮细胞中能高效表达,可使细胞增殖能力达0.301±0.096 (P<0.05)。结论CNP具有促进血管内皮细胞增殖的生物学活性。以聚乙烯亚胺介导转染质粒、检测CNP在血管内皮细胞中的表达及测定被转染细胞增殖活性等方法的建立,为开展CNP基因治疗提供了物质基础和实验依据。

To investigate the influence of human C-type natriuretic peptide （hCNP） on proliferation of vascular endothelial cells （HUVECs）. Methods Reconstructed pcDNA3. 1 （ ＋ ）/hCNP was transfected into HUVECs with polyethylenimine and its plasmid expression was examined with RT-PCR, immunohistochemistry and Western blot. MTT method was used to determine the effect of expressed protein on proliferation of HUVECs. pcDNA3.1 （ ＋ ）/hCNP transfection was used for control. Results The proliferation of HUVEC 48 h after pcDNA3.1 （ ＋ ）/hCNP transfection was （0. 301±0. 096） ,which was obvi- ously higher than that with pcDNA3. 1 （ ＋ ） transfection （0. 164±0. 012） . Reconstructed pcDNA3. 1 （ ＋ ）/hCNP might be expressed in HUVECs effectively and its protein expression was capable of promoting HUVECs proliferation markedly. Conclusion The successive expression of reconstructed pcDNA3. 1 （ ＋ ）/hCNP and the promoting activity of its expressed protein on HUVECs lay the foundation potential therapeutic value of C-type natriuretic peptide.