携氧载吲哚菁绿纳米粒协同光—声动力对类风湿关节炎滑膜成纤维细胞的毒性作用研究

发布时间：2018-05-14 17:16

本文选题：类风湿关节炎 + 滑膜成纤维细胞　；参考：《重庆医科大学》2017年硕士论文

【摘要】：类风湿关节炎(Rheumatoid arthritis,RA)是一种病因不明的慢性、进展性、多关节受累的自身免疫病。滑膜细胞增生、炎症细胞浸润、滑膜血管翳生成及关节腔局部乏氧微环境共同促进RA疾病进展。虽然目前的治疗手段能使大多数患者的病情得到很大程度的改善,但仍有部分患者对目前的治疗反应不佳或病程反复,疾病持续进展,最终可能导致关节受损及功能障碍。光动力疗法和声动力疗法具有良好的靶向性和细胞杀伤能力,其联合治疗策略PSDT(Photo-sonodynamic therapy)可协同光化学效应及声机械效应而增加细胞杀伤效果。本研究基于液态氟碳化合物良好的携氧及液气相变能力、吲哚菁绿(Indocyanine green,ICG)的荧光示踪及光/声动力学特性、乳酸-羟基乙酸共聚物(Poly(dl-lactide-co-glycolic acid),PLGA)良好包载药物的特性,制备同时携载O2和ICG的PLGA纳米粒(OI-NPs),用激光1联合低强度超声激发OI-NPs为手段,对RA关节病变的滑膜成纤维细胞及乏氧微环境作为目标,体外探讨OI-NPs协同PSDT对滑膜成纤维细胞MH7A细胞株的细胞毒性作用及潜在的机制。实验发现:(1)低强度超声辐照游离ICG,对MH7A滑膜成纤维细胞能产生显著的细胞杀伤效应,同时大量活性氧生成,验证了ICG的声敏剂特性,活性氧的产生是其发挥作用的重要机制之一。(2)比较了包载全氟丙烷(Perfluoro-n-pentane,PFP)或全氟己烷(Perfluorohexane,PFH)及ICG的PLGA纳米粒的稳定性及发生液气相变条件,发现PFP携氧的PLGA纳米粒稳定性良好,激光激发相变所需能量较低,是更安全有效的多功能纳米粒,因此后续实验选择PFP作为氧气载体。(3)OI-NPs是ICG和O2的稳定高效载体,可显著提高ICG的稳定性,明显增加滑膜成纤维细胞对ICG的摄取量;激光联合低强度超声辐照OI-NPs,能协同产生显著的滑膜成纤维细胞细胞毒性及引起细胞凋亡,同时细胞内活性氧产率明显增加。综上,多功能纳米粒OI-NPs可有效携载ICG和O2,OI-NPs介导的PSDT对滑膜成纤维细胞能产生显著的细胞杀伤效应,活性氧产生是引起细胞毒性的重要机制。该研究可为OI-NPs协同光-声动力靶向治疗RA奠定基础,有望探索一种针对RA滑膜增殖病变靶向治疗的新方法。
[Abstract]:Rheumatoid arthritis (RA) is a chronic, progressive, multiarticular autoimmune disease with unknown etiology. Synovial cell proliferation, inflammatory cell infiltration, synovial pannus formation and local hypoxic microenvironment in articular cavity contribute to the progression of RA. Although the current treatment methods can greatly improve the condition of most patients, but there are still some patients with poor response to the current treatment or repeated course of disease, the disease continues to progress, which may eventually lead to joint damage and dysfunction. Photodynamic therapy and sonodynamic therapy have good targeting and cytotoxicity. The combination of photochemical and acousto-mechanical effects can increase the cytotoxicity of the cells by the combination of photochemical and acoustic-mechanical effects. Based on the good oxygen-carrying and liquid-gas transformation ability of liquid fluorocarbons, the fluorescence tracer and photoacoustic characteristics of indocyanine green indocyanine (ICG), and the properties of lactic acid-co-glycolic acid (PLGA) copolymers, PLGA nanoparticles carrying O _ 2 and ICG were prepared. Laser 1 combined with low-intensity ultrasound was used to excite OI-NPs. Synovial fibroblasts and hypoxic microenvironment of RA joint lesions were used as targets. To investigate the cytotoxicity of OI-NPs in combination with PSDT on synovial fibroblast MH7A cell line and its potential mechanism. It was found that low intensity ultrasound irradiation of ICG could induce significant cytotoxicity to MH7A synovial fibroblasts, and a large number of reactive oxygen species (Ros) were produced, which proved the characteristics of ICG as a sound sensitizer. The production of reactive oxygen species (Ros) is one of the important mechanisms of Ros. (2) the stability of PLGA nanoparticles loaded with perfluoro-n-pentanean (PFPFP) or perfluorohexane (PFHs) and ICG were compared, and the stability of PLGA nanoparticles loaded with PFP was found to be good. Laser excitation of phase transition requires less energy and is a more safe and effective multifunctional nanoparticles. Therefore, PFP is chosen as the oxygen carrier. The OI-NPs are stable and efficient carriers of ICG and O2, which can significantly improve the stability of ICG. The uptake of ICG by synovial fibroblasts was significantly increased, and the toxicity and apoptosis of synovial fibroblasts induced by laser combined with low intensity ultrasound irradiation were significantly increased, and the production rate of reactive oxygen species in the cells increased significantly. In conclusion, multifunctional nanoparticles OI-NPs can effectively carry ICG and O2OI-NPs mediated PSDT to synovial fibroblasts can produce significant cytotoxicity, reactive oxygen species production is an important mechanism of cytotoxicity. This study may lay a foundation for OI-NPs combined with photoacoustic targeting therapy for RA, and it is expected to explore a new method of targeted therapy for RA synovial proliferative lesions.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R593.22

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