Department of Pharmacy, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan, 430064, P. R. China.
Hubei Key Laboratory of Theory and Application Research of Liver and Kidney in Traditional Chinese Medicine, Affiliated Hospital of Hubei University of Chinese Medicine, Wuhan, P. R. China.
BMC Complement Med Ther. 2024 May 11;24(1):186. doi: 10.1186/s12906-024-04489-z.
Cepharanthin alone or in combination with glucocorticoid (GC) has been used to treat chronic immune thrombocytopenia (ITP) since the 1990s. Cepharanthine (CEP) is one of the main active components of Cepharanthin. The purpose of this study was to investigate the effects of CEP on GC pharmacodynamics on immune cells and analyse the possible action mechanism of their interactions.
Peripheral blood mononuclear cells (PBMCs), T lymphocytic leukemia MOLT-4 cells and daunorubicin resistant MOLT-4 cells (MOLT-4/DNR) were used to evaluate the pharmacodynamics and molecular mechanisms. Drug pharmacodynamics was evaluated by WST-8 assay. P-glycoprotein function was examined by rhodamine 123 assay. CD4CD25Foxp3 regulatory T cells and Th1/Th2/Th17 cytokines were detected by flow cytometry. P-glycoprotein expression and GC receptor translocation were examined by Western blot.
CEP synergistically increased methylprednisolone (MP) efficacy with the suppressive effect on the cell viability of PBMCs. 0.3 and 1 μM of CEP significantly inhibited P-glycoprotein efflux function of CD4 cells, CD8 cells, and lymphocytes (P<0.05). 0.033 μM of CEP also inhibited the P-glycoprotein efflux function in MOLT-4/DNR cells in a concentration-dependent manner (P<0.001). However, 0.033 μM of CEP did not influence P-glycoprotein expression. 0.03~0.3 μM of CEP significantly increased the GC receptor distribution from the cytoplasm to the nucleus in a concentration-dependent manner in MOLT-4/DNR cells. The combination did not influence the frequency of CD4, CD4CD25 and CD4CD25Foxp3 T cells or the secretion of Th1/Th2/Th17 cytokines from PBMCs. In contrast, CEP alone at 1 μM decreased the percentage of CD4 T cell significantly (P<0.01). It also inhibited the secretion of IL-6, IL-10, IL-17, TNF-α, and IFN-γ.
CEP synergistically promoted MP pharmacodynamics to decrease the cell viability of the mitogen-activated PBMCs, possibly via inhibiting P-glycoprotein function and potentiating GC receptor translocation. The present study provides new evidence of the therapeutic effect of Cepharanthin alone or in combination with GC for the management of chronic ITP.
自 20 世纪 90 年代以来，盐酸千金藤素（CEP）单独或与糖皮质激素（GC）联合用于治疗慢性免疫性血小板减少症（ITP）。CEP 是千金藤素的主要活性成分之一。本研究旨在探讨 CEP 对 GC 对免疫细胞药效学的影响，并分析其相互作用的可能作用机制。
使用外周血单个核细胞（PBMCs）、T 淋巴细胞白血病 MOLT-4 细胞和柔红霉素耐药 MOLT-4 细胞（MOLT-4/DNR）评估药效学和分子机制。通过 WST-8 测定法评估药物药效学。通过罗丹明 123 测定法检测 P-糖蛋白功能。通过流式细胞术检测 CD4+CD25+Foxp3+调节性 T 细胞和 Th1/Th2/Th17 细胞因子。通过 Western blot 检测 P-糖蛋白表达和 GC 受体易位。
CEP 与甲泼尼龙（MP）协同增强了对 PBMCs 细胞活力的抑制作用。0.3 和 1 μM 的 CEP 显著抑制了 CD4 细胞、CD8 细胞和淋巴细胞的 P-糖蛋白外排功能（P<0.05）。0.033 μM 的 CEP 还呈浓度依赖性抑制 MOLT-4/DNR 细胞的 P-糖蛋白外排功能（P<0.001）。然而，0.033 μM 的 CEP 不影响 P-糖蛋白表达。0.03~0.3 μM 的 CEP 显著增加了 MOLT-4/DNR 细胞中 GC 受体从细胞质向核内的分布，呈浓度依赖性。该组合不影响 PBMCs 中 CD4、CD4+CD25+和 CD4+CD25+Foxp3+T 细胞的频率或 Th1/Th2/Th17 细胞因子的分泌。相比之下，CEP 单独在 1 μM 时显著降低 CD4+T 细胞的百分比（P<0.01）。它还抑制了 IL-6、IL-10、IL-17、TNF-α 和 IFN-γ 的分泌。
CEP 协同促进 MP 药效学，降低丝裂原激活的 PBMCs 的细胞活力，可能通过抑制 P-糖蛋白功能和增强 GC 受体易位。本研究为盐酸千金藤素单独或与 GC 联合治疗慢性 ITP 提供了新的治疗效果证据。
