Institute of Clinical Pharmacology, Anhui Medical University, Hefei, 230032, China; Key Laboratory of Anti-Inflammatory and Immune Medicine, Ministry of Education, Hefei, 230032, China; Anhui Collaborative Innovation Centre of Anti-Inflammatory and Immune Medicine, Hefei, 230032, China.
Institute of Clinical Pharmacology, Anhui Medical University, Hefei, 230032, China; Key Laboratory of Anti-Inflammatory and Immune Medicine, Ministry of Education, Hefei, 230032, China; Anhui Collaborative Innovation Centre of Anti-Inflammatory and Immune Medicine, Hefei, 230032, China.
Biomed Pharmacother. 2019 Nov;119:109432. doi: 10.1016/j.biopha.2019.109432. Epub 2019 Sep 12.
Methotrexate (MTX) is a commonly used drug for the treatment of rheumatoid arthritis (RA) and it has been studied in RA resistance recently. P-glycoprotein (P-gp) is one of the important transporters that mediate MTX resistance. This study investigated the effect of Paeoniflorin-6'-O-benzene sulfonate (code: CP-25) in the resistance of P-gp-mediated MTX to RA.
Adjuvant arthritis (AA) was induced in rats via complete Freund's adjuvant. The experimental groups were divided into normal group; AA model group; monotherapy groups, including CP-25, MTX and dexamethasone; and CP-25 combined with MTX group. The expression of P-gp in synovial tissue was measured by western blot and histochemistry. Besides, P-gp high expression of human hepatoma cell line Bel7402/5-FU and Bel7402 were chose to study in MTX resistance and the function of P-gp was detected by Flow cytometry.
CP-25 had a good therapeutic effect on AA rats, significantly improved manifestations and reduced the expression of P-gp in synovial tissue, spleen medulla and small intestinal epithelial cells in the apical tissues of AA rats. In addition, CP-25 significantly inhibited the up-regulation of P-gp induced by TNF-α stimulation in synoviocytes. Furthermore, according to the accumulation and efflux of rhodamine 123 in Bel7402/5-FU resistant cells and Bel7402 sensitive cells, CP-25 could reverse the resistance of MTX in Bel7402/5-FU cells compared with Bel7402 cells, which was reflected by the reduced IC50 values of MTX. Further study indicated that CP-25 could decrease P-gp expression and inhibit P-gp function in Bel7402/5-FU cells.
CP-25 regulates the expression of P-gp and inhibits the function of P-gp, thereby improving the resistance of MTX.
甲氨蝶呤（MTX）是一种常用于治疗类风湿关节炎（RA）的药物，最近研究发现其在 RA 耐药性中起作用。P-糖蛋白（P-gp）是介导 MTX 耐药性的重要转运体之一。本研究探讨了芍药苷-6'-O-苯磺酸酯（代号：CP-25）对 P-gp 介导的 MTX 耐药性的 RA 的影响。
通过完全弗氏佐剂诱导大鼠佐剂关节炎（AA）。实验组分为正常组；AA 模型组；单药组，包括 CP-25、MTX 和地塞米松；CP-25 联合 MTX 组。通过 Western blot 和组织化学法测量滑膜组织中 P-gp 的表达。此外，选择 P-gp 高表达的人肝癌细胞系 Bel7402/5-FU 和 Bel7402 来研究 MTX 耐药性，并通过流式细胞术检测 P-gp 的功能。
CP-25 对 AA 大鼠有良好的治疗作用，显著改善了大鼠的临床表现，并降低了 AA 大鼠滑膜组织、脾髓和小肠上皮细胞顶区 P-gp 的表达。此外，CP-25 显著抑制了 TNF-α刺激引起的滑膜细胞中 P-gp 的上调。此外，根据 rhodamine 123 在 Bel7402/5-FU 耐药细胞和 Bel7402 敏感细胞中的积累和外排，CP-25 可以降低 Bel7402/5-FU 细胞中 MTX 的 IC50 值，从而逆转 MTX 在 Bel7402/5-FU 细胞中的耐药性，与 Bel7402 细胞相比。进一步研究表明，CP-25 可以降低 Bel7402/5-FU 细胞中 P-gp 的表达并抑制 P-gp 的功能。
CP-25 调节 P-gp 的表达并抑制 P-gp 的功能，从而提高 MTX 的耐药性。
