Yao Li, Yang Yu-Xia, Cao Hui, Ren Huan-Huan, Niu Zheng, Shi Ling
Department of Medicinal Chemistry and Natural Medicine Chemistry, Department of Pharmacognosy, College of Pharmacy, Harbin Medical University, Harbin 150081, China; State-Province Key Laboratory of Biomedicine-Pharmaceutics of China, Harbin Medical University, Harbin 150081, China.
Department of Medicinal Chemistry and Natural Medicine Chemistry, Department of Pharmacognosy, College of Pharmacy, Harbin Medical University, Harbin 150081, China.
Chin J Nat Med. 2020 Apr;18(4):308-320. doi: 10.1016/S1875-5364(20)30038-8.
Osthole is observed to have the capacity to treat pulmonary arterial hypertension (PAH) in rats, but molecular mechanism is still unknown. The present study aims to discover therapeutic targets and explore therapeutic mechanism of osthole against PAH from metabolic perspective. A rat model with PAH was successfully established with MCT, following osthole administration, then untargeted metabolomics assay was performed using UPLC-Q-TOF-MS to identify differential metabolites and associated metabolic pathways, at last mechanism investigation was done by qRT-PCR, Western blot and ELISA. Differential metabolites characterized in rats with PAH were mostly assigned to sphingolipid metabolism, synthesis of unsaturated fatty acids, glycolysis, nucleotide metabolism, steroid hormone biosynthesis. Furthermore, osthole reversed high level of S1P by modulating metabolic enzyme Sphk1 in rats with PAH. In addition, osthole inhibited the expression of Sphk1 by downregulating microRNA-21, phosphorylation of Akt, phosphorylation of mTOR in vivo and in vitro. These results demonstrated that metabolomics is a promising approach to discover potential drug target for PAH treatment. Importantly, our findings further elucidated therapeutic mechanism of osthole, a natural product, having a role of metabolic regulator to potentially treat PAH by targeting inhibition of Sphk1/S1P via microRNA-21-PI3K/Akt/mTOR signal pathway. Altogether, this discovery paves a critical foundation for enabling osthole to be a candidate compound to treat PAH.
蛇床子素被观察到具有治疗大鼠肺动脉高压（PAH）的能力，但其分子机制尚不清楚。本研究旨在从代谢角度发现蛇床子素治疗PAH的靶点并探索其治疗机制。用野百合碱成功建立PAH大鼠模型，给予蛇床子素后，采用超高效液相色谱-四极杆-飞行时间质谱联用仪（UPLC-Q-TOF-MS）进行非靶向代谢组学分析，以鉴定差异代谢物及相关代谢途径，最后通过实时定量聚合酶链反应（qRT-PCR）、蛋白质免疫印迹法（Western blot）和酶联免疫吸附测定（ELISA）进行机制研究。PAH大鼠中特征性的差异代谢物大多与鞘脂代谢、不饱和脂肪酸合成、糖酵解、核苷酸代谢、类固醇激素生物合成有关。此外，蛇床子素通过调节PAH大鼠体内的代谢酶鞘氨醇激酶1（Sphk1）来逆转高水平的1-磷酸鞘氨醇（S1P）。此外，蛇床子素在体内外均通过下调微小RNA-21、蛋白激酶B（Akt）的磷酸化、哺乳动物雷帕霉素靶蛋白（mTOR）的磷酸化来抑制Sphk1的表达。这些结果表明，代谢组学是发现PAH治疗潜在药物靶点的一种有前景的方法。重要的是，我们的研究结果进一步阐明了天然产物蛇床子素的治疗机制，它作为一种代谢调节剂，可能通过微小RNA-21-磷脂酰肌醇3激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白信号通路靶向抑制Sphk1/S1P来治疗PAH。总之，这一发现为蛇床子素成为治疗PAH的候选化合物奠定了关键基础。
