NO缓释纳米粒子PEG-b-PAASNO对肺动脉平滑肌细胞的影响-东南大学学报医学版

NO缓释纳米粒子PEG-b-PAASNO对肺动脉平滑肌细胞的影响

单位：1. 东南大学附属中大医院呼吸与危重症医学科, 江苏南京 210009;
2. 中国科学院大学温州研究所, 浙江温州 325000

分类号：R543.2

出版年·卷·期（页码）：2024·43·第一期（1-8）

摘要：

目的: 探讨一氧化氮(NO)供体聚合物胶束PEG-b-PAASNO(PSNO)对肺动脉平滑肌细胞(PASMC)的影响。方法: 构建PSNO并检测体外释放NO水平。分离并培养大鼠PASMC,CCK8确定PSNO安全浓度范围和有效作用浓度。设立空白对照组、PSNO(250 ng·mL^-1)组(PSNO组)、血小板衍生生长因子-BB(PDGF-BB,15 ng·mL^-1)组(BB组)和PDGF-BB+PSNO(15 ng·mL^-1 PDGF-BB+250 ng·mL^-1 PSNO)组(BB+PSNO组),用5-乙炔基-2'脱氧尿嘧啶核苷(EDU)实验、划痕实验和Transwell实验评估PSNO对PASMC增殖、迁移能力的影响,免疫印迹法评估PASMC增殖、收缩、细胞周期相关蛋白[增殖细胞核抗原(PCNA)、钙调蛋白(calponin)、α-平滑肌肌动蛋白(α-SMA)、细胞周期蛋白D1(cyclin D1)、周期蛋白依赖性激酶2(CDK2)]的表达水平。组间比较采用one-way ANOVA分析。结果: 成功构建稳定释放NO的纳米聚合物胶束PSNO,连续释放NO超过48 h,效应呈时间-剂量依赖性。PSNO质量浓度为250 ng·mL^-1时有效抑制PDGF-BB诱导的PASMC增殖、迁移;与BB组相比,BB+PSNO组PCNA、calponin、α-SMA、cyclin D1和CDK2蛋白表达水平降低,差异均有统计学意义(P<0.05)。结论: 成功构建一种可以稳定释放NO的纳米聚合物胶束PSNO,其有效释放NO超48h,功能上阻滞PASMC增殖、迁移,并降低PASMC收缩、细胞周期相关蛋白表达水平。

Objective: To study the effect of nitric oxide(NO) donor nano-polymer micelle PEG-b-PAASNO(PSNO) on pulmonary artery smooth muscle cells(PASMC). Methods: PSNO was constructed and its release rate was assayed in vitro. Rat PASMCs were isolated and cultured. The safe concentration range and effective concentration of PSNO were determined by CCK8.The cells were divided into control group,PSNO(250 ng·mL^-1)group,platelet-derived growth factor-BB(PDGF-BB,15 ng·mL^-1) group(BB group),and PDGF-BB+PSNO(15 ng·mL^-1 PDGF-BB,250 ng·mL^-1 PSNO) group(BB+PSNO group). 5-ethynyl-2'-deoxyuridine(EDU),Transwell and wound healing were used to examine the effects of PSNO on the proliferation and migration ability of PASMC.The expression levels of proliferation,contraction and cell cycle-related proteins[proliferating cell nuclear antigen(PCNA),calponin,alpha-smooth muscle actin(α-SMA),cyclin D1and cyclin-dependent kinase 2(CDK2)] of PASMC were evaluated by Western blotting. Data were analyzed by one-way ANOVA analysis. Results: A nano-polymer micelle with stable NO release(PSNO) was constructed,and NO was continuously released for more than 48 h in a time-dose-dependent manner. When the concentration of PSNO was 250 ng·mL^-1,proliferation and migration of PASMC induced by PDGF-BB were effectively inhibited.Compared with BB group,the expression levels of PCNA,Calponin,α-SMA,cyclin D1 and CDK2 in BB+PSNO group were significantly lower(P<0.05).Conclusion: A nano-polymer micelle with stable release of NO(PSNO) is successfully constructed,which can effectively release NO for more than 48 h,blocking the proliferation and migration of PASMC,and reducing the expression level of PASMC contraction and cell cycle-related proteins.

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