Lu Yingyu, Chen Yonger, Li Yuhua, Xu Shuoxi, Lian Dawei, Liang Jian, Jiang Dongxu, Chen Shuxian, Hou Shaozhen
School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, PR China.
Department of Hepatobiliary Surgery, The Third Affiliated Hospital of Sun Yat-Sen University, Guangzhou, Guangdong 510000, PR China.
Int Immunopharmacol. 2023 Nov;124(Pt A):110838. doi: 10.1016/j.intimp.2023.110838. Epub 2023 Aug 24.
Colorectal cancer (CRC) is a growing concern due to its high morbidity and mortality, and the search for effective and less toxic active substances against inflammatory bowel diseases has been a hot topic in the research and development of drugs against CRC. It is reported that monotropein isolated from the roots of Morinda officinalis, can improve Dextran Sodium Sulfate (DSS)-induced ulcerative colitis in mice, but its therapeutic effects and mechanisms for CRC treatment are still to be investigated. In the present study, we first used molecular docking, BLI, CESTA, and DARTS methods to detest whether monotropein targets VDR proteins. In addition, we used tumor cell conditioned co-culture and four models of macrophage polarisation to investigate the regulation of four macrophage polarisations by monotropein using RT-PCR, IF and western blot. Furthermore, we further validated the target of action of monotropein for the treatment of Azoxymethane (AOM)/DSS induced colitis associated cancer (CAC) using knockout animals. Meanwhile, we further explored the mechanism of action of monotropein in regulating polarisation by detecting JAK/STAT1-related genes and proteins. Molecular docking and biofilm interference techniques showed that monotropein bound to the VDR, and additional results from CESTA and DARTS suggested that VDR proteins are targets of monotropein. Furthermore, in tumor cell conditioned co-cultures or LPS + IFN-γ induced RAW264.7 cells, VDR translocation to the nucleus was reduced, JAK1/STAT1 signaling pathway proteins were up-regulated, and macrophages were polarised towards the M1-type after monotropein intervention. Animal models in which normal VDR or myeloid VDR was knocked out confirmed that JAK1 levels in intestinal tissues were increased after monotropein intervention, macrophages were polarised towards the M1 type, and CAC paracarcinomas were ameliorated. Taken together, the present study concluded that monotropein inhibited colitis-associated cancers through macrophage polarisation regulated by VDR/JAK1/STAT1.
由于结直肠癌（CRC）的高发病率和死亡率，其日益受到关注，而寻找针对炎症性肠病的有效且毒性较小的活性物质一直是CRC药物研发中的热门话题。据报道，从巴戟天根部分离出的水晶兰苷可改善葡聚糖硫酸钠（DSS）诱导的小鼠溃疡性结肠炎，但其对CRC治疗的疗效和机制仍有待研究。在本研究中，我们首先使用分子对接、生物层干涉技术（BLI）、化学交换饱和转移（CESTA）和药物亲和反应靶标稳定性（DARTS）方法来检测水晶兰苷是否靶向维生素D受体（VDR）蛋白。此外，我们使用肿瘤细胞条件共培养和四种巨噬细胞极化模型，通过逆转录聚合酶链反应（RT-PCR）、免疫荧光（IF）和蛋白质免疫印迹法来研究水晶兰苷对四种巨噬细胞极化的调节作用。此外，我们使用基因敲除动物进一步验证了水晶兰苷治疗氧化偶氮甲烷（AOM）/DSS诱导的结肠炎相关癌症（CAC）的作用靶点。同时，我们通过检测与Janus激酶/信号转导和转录激活因子1（JAK/STAT1）相关的基因和蛋白，进一步探索了水晶兰苷调节极化的作用机制。分子对接和生物膜干涉技术表明水晶兰苷与VDR结合，CESTA和DARTS的其他结果表明VDR蛋白是水晶兰苷的靶点。此外，在肿瘤细胞条件共培养或脂多糖（LPS）+γ干扰素（IFN-γ）诱导的RAW264.7细胞中，水晶兰苷干预后VDR向细胞核的转位减少，JAK1/STAT1信号通路蛋白上调，巨噬细胞向M1型极化。正常VDR或髓系VDR基因敲除的动物模型证实，水晶兰苷干预后肠道组织中JAK1水平升高，巨噬细胞向M1型极化，CAC旁癌得到改善。综上所述，本研究得出结论，水晶兰苷通过VDR/JAK1/STAT1调节的巨噬细胞极化抑制结肠炎相关癌症。
