School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang, 110016, China.
Preparation Center, Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang, 110000, PR China.
Phytomedicine. 2023 Jul;115:154776. doi: 10.1016/j.phymed.2023.154776. Epub 2023 Mar 18.
The multi-drug resistance is an inherent weakness in the chemotherapeutics of non-small cell lung cancer occurring frequently all over the world. Clinically, ginseng and Chinese medicinal prescriptions including ginseng usually used as anti-tumor adjuncts due to its characteristic of qi-invigorating, which could improve the curative effect of chemotherapy drugs and reduce their toxic side effects. Triterpenoid saponins are the crucial active ingredients in Panax ginseng, and Ginsenoside Rb is of the highest quantities. However, the research on the tumor drug-resistance reversal effect and mechanism of ginsenoside Rb is still not clear.
This study aimed to systematically estimate the reversal activity of Ginsenoside Rb on cisplatin-insensitivity of A549/DDP cells and to reveal its prospective molecular mechanism.
MTT assay were conducted to evaluate the reversal activity on cisplatin-insensitivity of A549/DDP cells of Ginsenoside Rbin vitro, and the behavior was also studied by establishing a subcutaneous transplanted tumor model of A549/DDP in BALB/c-nu mice. In addition, P-gp ATPase activity assay, cisplatin accumulation assay, Annexin V-FITC apoptosis assay, real-time qPCR analysis and western blotting analysis were used to clarify the potential mechanism.
Ginsenoside Rb could effectively reverse the cisplatin-resistance of A549/DDP in vitro and vivo. And after the co-treatment of Ginsenoside Rb plus cisplatin, the accumulation of cisplatin increased in A549/DDP cells, which was accompanied with the down-regulation of the mRNA and protein expression levels of ABCB1, SHH, PTCH1 and GLI2. Besides, the apoptosis-inducing ability of cisplatin improved by the relative regulation on the protein expression level of Bax and Bcl-2. Far more importantly, the changes of CYP3A4 mRNA and protein levels were not significant.
Ginsenoside Rb could increase the concentration of intracellular cisplatin and improve the insensitivity for cisplatin on A549/DDP cells. Even better, there was perhaps no unpredictable CYP3A4-mediated pharmacokinetic interactions after the combination of Ginsenoside Rb plus cisplatin. Ginsenoside Rb was a probable reversal agent for the cisplatin-insensitivity of A549/DDP cells, with a bifunction of inhibiting the efflux of two drug pumps (P-gp and PTCH1) by targeting ABCB1 and Hedgehog (Hh) pathway. In general, this research laid the groundwork for the development of a new reversal agent for the cisplatin-insensitivity of NSCLC.
多药耐药性是全球普遍存在的非小细胞肺癌化疗的固有弱点。临床上，人参和包括人参在内的中药方剂通常被用作抗肿瘤辅助药物，因为它具有补气的作用，可以提高化疗药物的疗效，降低其毒副作用。三萜皂苷是人参的关键活性成分，其中人参皂苷 Rb 的含量最高。然而，人参皂苷 Rb 逆转肿瘤耐药的作用及其机制的研究尚不清楚。
本研究旨在系统评估人参皂苷 Rb 对 A549/DDP 细胞顺铂耐药性的逆转作用，并揭示其潜在的分子机制。
采用 MTT 法体外评价人参皂苷 Rb 对 A549/DDP 细胞顺铂耐药性的逆转作用，并在 BALB/c-nu 小鼠建立 A549/DDP 皮下移植瘤模型进行体内研究。此外，还进行了 P-糖蛋白（P-gp）ATP 酶活性测定、顺铂蓄积测定、Annexin V-FITC 凋亡测定、实时 qPCR 分析和 Western blot 分析，以阐明潜在的机制。
人参皂苷 Rb 能有效逆转 A549/DDP 细胞的顺铂耐药性，体内外均有效果。并且，人参皂苷 Rb 联合顺铂处理后，A549/DDP 细胞内顺铂蓄积增加，同时 ABCB1、SHH、PTCH1 和 GLI2 的 mRNA 和蛋白表达水平下调。此外，通过对 Bax 和 Bcl-2 蛋白表达水平的相对调节，增强了顺铂的诱导凋亡能力。更重要的是，CYP3A4mRNA 和蛋白水平的变化不明显。
人参皂苷 Rb 能增加细胞内顺铂浓度，提高 A549/DDP 细胞对顺铂的敏感性。更好的是，人参皂苷 Rb 联合顺铂后，可能不存在不可预测的 CYP3A4 介导的药代动力学相互作用。人参皂苷 Rb 可能是 A549/DDP 细胞顺铂耐药性的一种潜在逆转剂，通过靶向 ABCB1 和 Hedgehog（Hh）通路抑制两种药物泵（P-gp 和 PTCH1）的外排作用。总的来说，这项研究为开发 NSCLC 顺铂耐药性的新型逆转剂奠定了基础。
