Faculty of Pharmacy, Hubei University of Chinese Medicine, Wuhan, 430065, China.
Faculty of Pharmacy, Hubei University of Chinese Medicine, Wuhan, 430065, China; Key Laboratory of Traditional Chinese Medicine Resources and Chemistry of Hubei Province, Wuhan, 430061, China.
J Ethnopharmacol. 2021 Oct 5;278:114267. doi: 10.1016/j.jep.2021.114267. Epub 2021 Jun 1.
As an important Chinese herb, Coptis chinensis Franch. (Huanglian, HL) has a long history of usage for clearing heat, eliminating dampness, purging fire and detoxification in Traditional Chinese Medicine (TCM). HL, also called goldthread, was frequently used for the treatment of typhoid, tuberculosis, epidemic cerebrospinal meningitis, pertussis, and other lung-related diseases. Modern research has shown that HL and its main compounds also have anti-tumor effects. However, studies have not reported whether its main compounds inhibit Non-small cell lung cancer (NSCLC) development and progression.
This study aimed to find out the potential targets and mechanisms of Oxyepiberberine (OPB) isolated from HL in the treatment of NSCLC, using network pharmacology and biological experimental.
Silica gel chromatography column was used to isolate OPB from HL, and the structure of OPB was elucidated using different spectroscopic analysis methods, including H-nuclear magnetic resonance (NMR), C-NMR and electrospray ionization mass spectrometry (ESI/MS). MTT assay was performed to determine cell proliferation of OPB on A549, H1975 and BEAS-2B cells. Then, the potential targets, pathways and hub genes of OPB for treating NSCLC were screened out through network pharmacology. Based on the results of network pharmacology, core targets of OPB for treating NSCLC were docking with OPB via molecular docking. Wound healing, plate clone, Hoechst staining, and western blot assay were used to verify the function of OPB in treatment of NSCLC.
OPB was isolated from the HL, its molecular formula was identified as CHNO. Through MTT, OPB significantly inhibited the proliferation of H1975 cells and A549 cells, and A549 was chosen as the test cancer cell. Through network pharmacology, 22 potential targets, 156 related-pathways, and 6 hub genes were screened out. The results of molecular docking showed that SRC, BRAF, and MMP9 were the core targets of OPB against NSCLC. Through biological experimental, it was found that OPB inhibited growth and migration of A549 cells. In addition, OPB induced apoptosis in A549 cells. Through western blot assay, the expressions of Src, ERK1/2 and other four proteins were down-regulated, which suggested that OPB inhibited the proliferation of lung cancer cells by down-regulating SRC-FAK-RAS-RAF-MEK-ERK pathway, so as to achieve the anti-NSCLC effect.
Our study demonstrated that anti-NSCLC effect of OPB through network and experiments, which provided a theoretical basis for the clinical antitumor of OPB, and provided a foundation for further study of OPB.
黄连（HL）作为一种重要的中国草药，在中药中具有清热燥湿、泻火解毒的悠久历史。HL，也称为金线，常用于治疗伤寒、肺结核、流行性脑脊髓膜炎、百日咳和其他肺部相关疾病。现代研究表明，HL 及其主要化合物也具有抗肿瘤作用。然而，研究尚未报道其主要化合物是否抑制非小细胞肺癌（NSCLC）的发展和进展。
本研究旨在通过网络药理学和生物学实验，找出 HL 中分离得到的小檗碱（OPB）治疗 NSCLC 的潜在靶点和机制。
硅胶柱层析法从 HL 中分离 OPB，采用不同的光谱分析方法（包括 H-核磁共振（NMR）、C-NMR 和电喷雾电离质谱（ESI/MS））对 OPB 的结构进行解析。采用 MTT 法检测 OPB 对 A549、H1975 和 BEAS-2B 细胞增殖的影响。然后，通过网络药理学筛选出 OPB 治疗 NSCLC 的潜在靶点、途径和关键基因。基于网络药理学的结果，采用分子对接法将 OPB 与 OPB 进行对接。划痕愈合、平板克隆、Hoechst 染色和 Western blot 检测用于验证 OPB 治疗 NSCLC 的功能。
从 HL 中分离出 OPB，其分子式鉴定为 CHNO。通过 MTT，OPB 显著抑制 H1975 细胞和 A549 细胞的增殖，选择 A549 作为测试癌细胞。通过网络药理学筛选出 22 个潜在靶点、156 个相关通路和 6 个关键基因。分子对接结果表明，SRC、BRAF 和 MMP9 是 OPB 治疗 NSCLC 的核心靶点。通过生物学实验发现，OPB 抑制 A549 细胞的生长和迁移。此外，OPB 诱导 A549 细胞凋亡。通过 Western blot 检测，下调 Src、ERK1/2 等四种蛋白的表达，提示 OPB 通过下调 SRC-FAK-RAS-RAF-MEK-ERK 通路抑制肺癌细胞增殖，从而达到抗 NSCLC 的作用。
本研究通过网络和实验证明了 OPB 对 NSCLC 的抗作用，为 OPB 的临床抗肿瘤提供了理论依据，为 OPB 的进一步研究奠定了基础。
