Xia Donghai, Jiang Xuan, Xie Xiaomin, Zhou Han, Yu Dongping, Jin Gaowa, Ye Xianlong, Zhu Shenglong, Guo Zhimou, Liang Xinmiao
Key Laboratory of Phytochemistry and Natural Medicines, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
Int J Mol Sci. 2025 Mar 20;26(6):2793. doi: 10.3390/ijms26062793.
Danshen, a well-known traditional Chinese medicine (TCM), has gained increasing attention for its protective effects on nonalcoholic fatty liver disease (NAFLD). However, the molecular mechanisms underlying these effects remain to be elucidated. Niemann-Pick C1-like 1 (NPC1L1), a key transporter mediating intestinal cholesterol absorption, has emerged as a critical target for NAFLD treatment. This study aimed to screen for NPC1L1 inhibitors from Danshen and investigate their therapeutic effects on NAFLD. We established a high-throughput screening platform using stable Caco2 cell lines expressing human NPC1L1 (hL1-Caco2) and discovered that tanshinones (Tans), the liposoluble components of Danshen, inhibited NPC1L1-mediated cholesterol absorption in hL1-Caco2 cells. Additionally, Tans treatment reduced hepatic steatosis in high-fat diet (HFD)-fed mice. To identify the active compounds in Tans, activity-oriented separation was performed by integrating the high-throughput screening platform and two-dimensional chromatographic techniques. Ultimately, cryptotanshinone (CTS) was identified as a novel NPC1L1 inhibitor and significantly decreased hepatic steatosis in HFD-fed mice. Molecular docking and dynamics simulation showed that CTS stably bound with NPC1L1, where TRP383 acted as the key amino acid. Taken together, this study demonstrates, for the first time, that CTS, a liposoluble compound from Danshen, is a novel NPC1L1 inhibitor. Our findings suggest that the inhibitory effect of CTS against NPC1L1-mediated intestinal cholesterol absorption may be a potential mechanism, contributing to its alleviation of NAFLD in mice.
丹参是一种著名的传统中药，其对非酒精性脂肪性肝病（NAFLD）的保护作用日益受到关注。然而，这些作用背后的分子机制仍有待阐明。尼曼-匹克C1样1蛋白（NPC1L1）是介导肠道胆固醇吸收的关键转运蛋白，已成为NAFLD治疗的关键靶点。本研究旨在从丹参中筛选NPC1L1抑制剂，并研究其对NAFLD的治疗作用。我们利用表达人NPC1L1的稳定Caco2细胞系（hL1-Caco2）建立了一个高通量筛选平台，发现丹参的脂溶性成分丹参酮（Tans）可抑制hL1-Caco2细胞中NPC1L1介导的胆固醇吸收。此外，丹参酮处理可减轻高脂饮食（HFD）喂养小鼠的肝脂肪变性。为了鉴定丹参酮中的活性化合物，通过整合高通量筛选平台和二维色谱技术进行了活性导向分离。最终，隐丹参酮（CTS）被鉴定为一种新型NPC1L1抑制剂，并显著降低了HFD喂养小鼠的肝脂肪变性。分子对接和动力学模拟表明，CTS与NPC1L1稳定结合，其中色氨酸383是关键氨基酸。综上所述，本研究首次证明，丹参中的脂溶性化合物CTS是一种新型NPC1L1抑制剂。我们的研究结果表明，CTS对NPC1L1介导的肠道胆固醇吸收的抑制作用可能是其减轻小鼠NAFLD的潜在机制。
