银屑灵优化方干预miR-31介导角质细胞代谢重组的机制研究

Cell metabolic reprogramming plays a key role in change of biological behavior of keratinocytes in psoriasis, and importantly, miR-31 critically promotes metabolic reprogramming in keratinocytes. PSORI-CM was originated from clinical experience of a Chinese medicine master, and long-term clinical research and practice have confirmed the curative effect of this compound Chinese medicine. Moreover, experimental studies found that PSORI-CM has many effects in anti-inflammation, immune regulation, as well as inhibiting proliferation and differentiation of keratinocytes. However, mechanisms underlying effects of PSORI-CM are still under investigation. We have previously found that PSORI-CM could significantly inhibit the expression of miR-31 in psoriatic skin lesions of psorisiform mouse model. In addition, research results of network pharmacology revealed that PSORI-CM perhaps target the key pathways involved in cell metabolic reprogramming. These data suggest that PSORI-CM may regulate the pathological phenotype of psoriatic keratinocytes through targeting miR-31-mediated metabolic reprogramming. Therefore, we hypothesize that intervention of miR-31 mediated metabolic reprogramming in keratinocyte by PSORI-CM. In this project, cellular metabolisms, protein-chip, signaling studies and other technical means will be employed , and in vivo and in vitro models will be combined, from the macroscopic and microcosmic angle, to study and delineated the "miR-31-keratinocytes metabolic reprogramming- keratinocytes pathological phenotype" mechanism roadmap for PSORI-CM effects. This study will to some extent reveal the scientific connotation the effect of PSORI-CM in the future.

细胞代谢重组是银屑病角质细胞生物学行为改变的关键因素，而miR-31是银屑病细胞代谢重组的关键分子。银屑灵优化方源于国医大师经验方，长期临床实践及临床研究证实其疗效肯定，实验研究发现该方具有较好的抗炎及抗角质细胞增殖与分化作用，但是其“标本兼治”的科学内涵仍有待进一步阐明。项目组前期研究显示该方可抑制银屑病动物模型皮损处miR-31的表达，网络药理学研究揭示该方靶向细胞代谢重组的关键通路，提示该方可能通过靶向miR-31介导的角质细胞代谢重组，从而改变银屑病角质细胞病理表型，发挥治疗作用。因此，本项目提出“银屑灵优化方干预miR-31介导角质细胞代谢重组”的科学假说，利用细胞代谢组学、蛋白芯片、信号通路研究等技术手段，动物模型和体外细胞模型研究相互结合探讨银屑灵优化方干预“miR-31—细胞代谢重组—角质细胞病理表型”的作用机制图谱，从而阐释其“标本兼治”的科学内涵。

细胞代谢重组是银屑病主要靶器官角质细胞生物学行为改变的关键因素，而作为潜在银屑病标志物的miR-31不仅参与皮损处角质细胞病理表型改变，还是银屑病细胞代谢重组的关键分子。本课题应用了包括代谢组、同位素示踪代谢流、SILAC蛋白质组、DIA蛋白质组、Seahorse、Luminex等前沿技术，深入探索关键分子miR-31介导的角质细胞代谢重编程以及银屑灵优化方调控角质细胞代谢重编程的具体分子机制。主要研究发现包括：（1）创新性的开发基于SILAC蛋白质组结合基因种子序列筛选miR-31的直接靶标，可拓展至其他miRNA的靶标鉴定研究；（2）揭示miR-31介导角质细胞代谢重编程的具体分子机制，为认识银屑病皮损角质细胞的代谢特征提供参考并为开发靶向细胞代谢的治疗新策略提供思路；（3）发现miR-31过表达的角质细胞可与naïve CD4+T细胞发生依赖于天冬氨酸的代谢交互，促进Th17细胞分化，从而加剧银屑病病理进展；（4）基于临床样本揭示银屑病皮损组织代谢基因的表达谱；（5）揭示银屑病患者外周血中高表达天冬氨酸为特征的代谢图谱；（6）从细胞代谢重编程角度，阐明银屑灵优化方靶向mTOR/HK2/glycolysis轴抑制角质细胞增殖的机制；（7）采用液相色谱串联质谱测定法鉴定23个银屑灵优化方的入血成分，为后续针对该方的机制和物质基础研究提供基础。本课题的主要研究成果不仅解答课题的关键科学问题，达到预期目标，而且促进了中医药防治银屑病在细胞代谢重编程层面的研究，为各种前沿技术在中医药领域的科学研究提供参考。目前课题组已发表SCI论文2篇，其中1篇发表于《EMBO molecular medicine》（中科院1区，IF:14.26），另外已投稿SCI论文1篇（返修状态），发表中文核心期刊2篇，获国家发明专利授权2项，培养国家中医药传承创新团队负责人1名，岐黄学者1名，博士研究生4名，硕士研究生2名，做国内大会报告1次，取得较好的科研成果。