School of Pharmacy, Shenyang Pharmaceutical University, Wenhua Road 103, Shenyang, Liaoning 110016, China.
Mailman Research Center, McLean Hospital, Harvard Medical School, Boston, MA, United States.
Phytomedicine. 2021 Dec;93:153813. doi: 10.1016/j.phymed.2021.153813. Epub 2021 Oct 19.
The bioactive alkaloids identified from Cortex Phellodendri (CP) were highly effective in treating rats with benign prostatic hyperplasia (BPH). Specifically, lipoxygenase-5 (LOX-5) and cyclooxygenase-2 (COX-2) were identified as two primary targets for alleviating inflammation in BPH rats. However, it remains unknown whether the alkaloid components in CP can interact with the two target proteins.
To further identify bioactive alkaloids targeting LOX/COX pathways.
An affinity-ultrafiltration mass spectrometry approach was employed to screen dual-target LOX-5/COX-2 ligands from alkaloid extract. The structures of bioactive alkaloids were characterized by high-resolution Fourier transform ion cyclotron resonance mass spectrometry. To understand the molecular mechanisms underlying the effects of bioactive alkaloids, the expression levels of LOX-5 and COX-2 in BPH model rats were investigated at both protein and mRNA levels. The LOX-5/COX-2 enzymes activity experiments and molecular docking analysis were performed to fully evaluate the interactions between bioactive alkaloids and LOX-5/COX-2.
After comprehensive analysis, the results showed that bioactive alkaloids could suppress the expression of LOX-5 and COX-2 simultaneously to exert an anti-inflammatory effect on the progression of BPH. In addition, the screened protoberberine, demethyleneberberine was found to exhibit prominent inhibitory activities against both LOX-5 and COX-2 enzymes, palmatine and berberine with moderate inhibitory activities. Molecular docking analysis confirmed that demethyleneberberine could interact well with LOX-5/COX-2.
This study is the first to explore the inhibitory effects of bioactive alkaloids from CP on LOX-5 and COX-2 activities in BPH rats. Our findings demonstrate that the bioactive alkaloids from CP can ameliorate BPH via dual LOX-5/COX-2 pathways, which serves as an efficient approach for the discovery of novel drug leads from natural products with reduced side effects.
从黄柏（CP）中鉴定出的生物活性生物碱在治疗良性前列腺增生（BPH）大鼠方面非常有效。具体来说，脂氧合酶-5（LOX-5）和环氧化酶-2（COX-2）被确定为缓解 BPH 大鼠炎症的两个主要靶点。然而，目前尚不清楚 CP 中的生物碱成分是否可以与这两个靶蛋白相互作用。
进一步鉴定靶向 LOX/COX 途径的生物活性生物碱。
采用亲和超滤质谱法从生物碱提取物中筛选双靶标 LOX-5/COX-2 配体。通过高分辨率傅里叶变换离子回旋共振质谱对生物活性生物碱的结构进行了表征。为了了解生物活性生物碱作用的分子机制，在蛋白质和 mRNA 水平上研究了 BPH 模型大鼠中 LOX-5 和 COX-2 的表达水平。进行 LOX-5/COX-2 酶活性实验和分子对接分析，以充分评估生物活性生物碱与 LOX-5/COX-2 的相互作用。
经过综合分析，结果表明生物活性生物碱可同时抑制 LOX-5 和 COX-2 的表达，从而对 BPH 的进展发挥抗炎作用。此外，筛选出的原小檗碱、去亚甲基小檗碱对 LOX-5 和 COX-2 酶均表现出显著的抑制活性，黄连碱和小檗碱表现出中等抑制活性。分子对接分析证实去亚甲基小檗碱可与 LOX-5/COX-2 良好相互作用。
本研究首次探讨了 CP 中的生物活性生物碱对 BPH 大鼠 LOX-5 和 COX-2 活性的抑制作用。我们的研究结果表明，CP 中的生物活性生物碱可通过双重 LOX-5/COX-2 途径改善 BPH，这为从天然产物中发现具有减少副作用的新型药物提供了一种有效的方法。
