HMGB1 A-box磁性纳米长循环脂质体的构建及其对实验性结肠炎的治疗作用

In the present study, the high mobility groupbox chromosomal protein 1(HMGB1),a key modulator in the complicated network of pre-inflammatory cytokines, was targeted with HMGB1 A-box, an HMGB1 analog modified with nanolipisomes and loaded with magnetic iron oxides,to enhance accumulation and extend the half of protein.The effectiveness of HMGB1 A-box was evaluated in mouse models of experimental colitis. HMGB1 A-box protein was obtained from stimulated RAW264.7 cells using a Ni2+ affinity column, and antiphase evaporation was performed on HMGB1.A-box proteins modified with magnetic nanolipisomes. The magnetism, form, particle size, drug-loading rate and body distribution of HMGB1 A-box were assayed, and parameters of pathological injury and pre-inflammatory cytokines were determined on models of colitis, which was triggered by OXZ or TNBS. The purpose of the project was to determine if HMGB1 A-box modified with magnetic nanolipisomes can be used as a targeting agent for inflammatory bowel disease.

本课题根据高迁移率族蛋白1(HMGB1)这一炎症性肠病等自身免疫性疾病炎症因子网络中的关键调控因子,设计利用它的拮抗物HMGB1 A-box,以纳米脂质体加载四氧化三铁等技术修饰,增加靶向聚集性能,延长半衰期,并观察评价其对实验性结肠炎的治疗作用。HMGB1 A-box蛋白由激发RAW264.7细胞株表达、Ni2+亲和层析柱纯化分离获取；采用逆相蒸发法分步制备磁性纳米长循环脂质体介导的HMGB1 A-box,并作磁性、形态、粒径和载药量等理化性质以及外磁场下体内分布等鉴定；制备噁唑酮和三硝基苯磺酸诱发的实验性结肠炎模型（Th2、Th1型免疫损伤），以HMGB1 A-box磁性纳米长循环脂质体干预，进行病理损伤指标和炎症因子的检测与分析，评价其是否具有治疗实验性结肠炎靶向生物制剂的研发前景。

本课题根据高迁移率族蛋白1(HMGB1)这一炎症性肠病等自身免疫性疾病炎症因子网络中的关键调控因子,设计利用它的拮抗物HMGB1 A-box,以纳米脂质体技术修饰,增加聚集性能,延长半衰期,并观察评价其对实验性结肠炎的治疗作用。HMGB1 A-box 蛋白的表达与纯化采用SUMO方法获得，该方法可高效、可溶性表达了重组HMGB1 A-box蛋白；采用薄膜-超声分散法分步制备纳米脂质体介导的HMGB1A-box,并作粒径和载药量等理化性质等鉴定；制备葡聚糖硫酸钠(DSS)实验性结肠炎模型，以HMGB1 A-box纳米脂质体干预，进行病理损伤指标和炎症因子的检测与分析，结果发现HMGB1 A-box纳米脂质体可减轻实验性结肠炎大鼠结肠组织炎症损伤，改善腹泻症状，下调HMGB1、TNF-α、IL-1、IL-6等细胞因子水平。本研究表明，HMGB1 A-box纳米脂质体有望成为新的抗实验性结肠炎药物分子。