Liver-targeted delivery of TSG-6 by calcium phosphate nanoparticles for the management of liver fibrosis
Liver-targeted delivery of TSG-6 by calcium phosphate nanoparticles for the management of liver fibrosis
复制标题
通过磷酸钙纳米颗粒肝脏靶向递送 TSG-6 以治疗肝纤维化
DOI：
10.7150/thno.37301
复制
发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Han, Ying
中科院分区：
文献类型：
--
作者：
Wang, Min;Zhang, Miao;Han, Ying
骨髓间充质干细胞（MSCs）移植是一种有前途的抗肝纤维化策略，但面临临床争议。受纳米医学进展的启发，我们的目的是通过确定骨髓间充质干细胞的关键抗纤维化分子和开发一种特异性的肝靶向nanocarrier.Methods：骨髓间充质干细胞分泌的细胞因子进行了检查与血清刺激的肝硬化患者绕过这些临床障碍。应用免疫组织化学、微阵列、免疫印迹和定量实时PCR（qRT-PCR）来鉴定关键的抗纤维化细胞因子并发现其在调节抗纤维化效应中的作用。采用生物矿化法制备磷酸钙纳米颗粒。通过CCl 4诱导的小鼠肝纤维化模型的体内成像和生化研究，评价纳米粒的靶向性和治疗效果。结果：经刺激后的骨髓间充质干细胞表达肿瘤坏死因子（TNF）刺激基因6（TSG-6）。在动物实验中，外源性给予单独的TSG-6可以改善肝纤维化，而TSG-6敲除的MSC（Lv-TSG-6 MSC）失去抗纤维化作用。进一步的研究证实了TSG-6的重要性，并通过调节M2巨噬细胞和增加基质金属蛋白酶12（MMP 12）表达来确定其抗纤维化机制。此外，我们发现了TSG-6，MMP 12和促炎细胞因子（TNF-α，IL-6和IL-1 β）之间的反馈回路，这可能会提高我们对肝硬化加重过程和TSG-6和MSC抗纤维化机制的理解。在此基础上，我们以牛血清白蛋白（BSA）为生物模板，通过生物矿化法制备了磷酸钙纳米颗粒（CaP@BSA NPs）。成像跟踪和药物负载研究显示了制备的CaP@BSA纳米粒的特异性肝靶向和高TSG-6负载功效。结论：TSG-6是骨髓间充质干细胞抗肝纤维化的主要细胞因子，TSG-6负载CaP @BSA纳米粒具有肝内特异性蓄积作用，可提高治疗效果，提示CaP@BSA有望成为肝脏疾病治疗的药物载体。
Mesenchymal stem cells (MSCs) transplantation is a promising antifibrotic strategy but facing clinical controversies. Inspired by advances in nanomedicine, we aimed to bypass these clinical barriers of MSCs by identifying the key antifibrotic molecule of MSCs and developing a specific liver-targeting nanocarrier.Methods: Cytokines secreted by MSCs were examined with serum stimulation of cirrhotic patients. Immunohistochemistry, microarray, immunoblotting, and quantitative real-time PCR (qRT-PCR) were applied to identify the critical antifibrotic cytokine and to discover its role in modulating antifibrotic effects. Biomineralization method was used to prepare calcium phosphate nanoparticles (NPs). The targeting and therapeutic efficiency of NPs were evaluated by in vivo imaging and biochemical studies on fibrotic mice induced by CCl4.Results: The stimulated MSCs exhibited high-level expression of Tumor necrosis factor (TNF)-stimulated gene 6 (TSG-6). On animal study, exogenous administration of TSG-6 alone can ameliorate liver fibrosis while TSG-6 knocked MSCs (Lv-TSG-6 MSCs) lost antifibrotic effects. Further studies verified the importance of TSG-6 and identified its antifibrotic mechanism by modulating M2 macrophages and increasing matrix metalloproteinase 12 (MMP12) expression. Additionally, we found a feedback loop between TSG-6, MMP12 and pro-inflammatory cytokines (TNF-alpha, IL-6, and IL-1 beta), which may improve our understanding of the aggravating process of cirrhosis and antifibrotic mechanisms of TSG-6 and MSCs. Based on these findings, we developed calcium phosphate nanoparticles (CaP@BSA NPs) by biomineralization method using bovine serum albumin (BSA) as the biotemplate. Imaging tracking and drug loading studies showed specific liver targeting and high TSG-6 loading efficacy of as-prepared CaP@BSA NPs. In vivo therapeutic study further demonstrated the improved therapeutic effects of TSG-6 loaded CaP@BSA.Conclusions: TSG-6 was a major antifibrotic cytokine of MSCs, TSG-6 loaded CaP@BSA NPs showed specific liver accumulation and improved therapeutic effects, which indicated translational potentials of CaP@BSA as a promising drug carrier for the liver disease management.