Bai Rui, Zhu Li-Zeng, Shao Changfa, Yin Zheng, Liu Qun, Gu Yu, Liu Bin
School of Materials Science and Engineering, Suzhou University of Science and Technology, Kerui Road, Suzhou, 215009, P. R. China.
State Key Laboratory of Natural Medicines, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing, 210009, P. R. China.
Adv Sci (Weinh). 2025 Aug;12(30):e02850. doi: 10.1002/advs.202502850. Epub 2025 May 23.
Liver fibrosis, a critical precursor to cirrhosis and a leading cause of mortality, highlights the urgent need for the identification of effective therapeutics. Activation of hepatic stellate cells (HSCs) is a key process in liver fibrosis. This study presents a live-cell drug-screening approach that specifically targets fibrosis-associated collagen type I alpha 1 (COL1A1) mRNA in activated HSCs through the use of fluorogenic RNA self-assembly. It employs a dual-probe system to construct an RNA Mango II structure, which upon binding with COL1A1 mRNA, facilitates activation of the TO1-Biotin fluorophore, thereby enabling the visualization of mRNA to indicate HSC activation levels. Through the application of a high throughput live-cell screening system, dihydrotanshinone I (DHT) is identified as potent leading antifibrotic compound, evidenced by its inhibitory effects on COL1A1 mRNA expression. The therapeutic efficacy of DHT is further substantiated by monitoring COL1A1 mRNA dynamics following treatment. In vivo studies demonstrates the sustained administration of DHT significantly ameliorated liver fibrosis in mice models. This method offers a simple, cost-effective approach of visualizing RNA dynamics and conducting drug screening in live cells, presenting a significant potential for the development of hepatic fibrosis therapies.
肝纤维化是肝硬化的关键前驱病变和主要死因，凸显了识别有效治疗方法的迫切需求。肝星状细胞（HSCs）的激活是肝纤维化的关键过程。本研究提出了一种活细胞药物筛选方法，该方法通过使用荧光RNA自组装，特异性地靶向活化HSCs中与纤维化相关的I型胶原α1（COL1A1）mRNA。它采用双探针系统构建RNA Mango II结构，该结构与COL1A1 mRNA结合后，促进TO1-生物素荧光团的激活，从而使mRNA可视化以指示HSC激活水平。通过应用高通量活细胞筛选系统，二氢丹参酮I（DHT）被鉴定为有效的抗纤维化先导化合物，这通过其对COL1A1 mRNA表达的抑制作用得到证明。通过监测治疗后COL1A1 mRNA的动态变化，进一步证实了DHT的治疗效果。体内研究表明，在小鼠模型中持续给予DHT可显著改善肝纤维化。该方法提供了一种在活细胞中可视化RNA动态和进行药物筛选的简单、经济有效的方法，为肝纤维化治疗的发展具有显著潜力。
