Dornand J, Favero J, Bonnafous J C, Mani J C
Cell Immunol. 1984 Feb;83(2):351-9. doi: 10.1016/0008-8749(84)90314-9.
At concentrations as low as 10(-7) M, the cardiotonic glycosteroid ouabain, a specific inhibitor of the membrane Na+, K+-ATPase, is known to inhibit in vitro human lymphocyte proliferation produced in mixed lymphocyte cultures or induced by various stimulating agents (PHA, Con A, PWM, soluble antigens), while mouse lymphocyte proliferation is unaffected at this concentration. Ouabain inhibits most of proliferative response parameters at all stages of the transformation. This observation prompted us to suggest that ouabain could also act through inhibition of interleukin production which is known to occur during the first hours after T-cell stimulation in the presence of monocytes. In order to check the possible influence of ouabain on interleukin production, conditioned media from stimulated human mononuclear cells, prepared in the presence or in the absence of inhibitor, were tested for their ability to promote a mouse thymocyte response to PHA. Instead of the expected inhibition, we found that ouabain, even at high concentrations (2 X 10(-6) M) enhanced the stimulatory effect and/or the production of murine thymocyte activating factor(s). Moreover conditioned media from serum-free cultures of unstimulated human mononuclear cells exposed for 24 hr to low ouabain concentrations (10(-8) to 10(-7) M) showed a high activating effect on the response of murine thymocytes to PHA. This soluble factor produced upon ouabain treatment is produced by adherent cells and appears to be functionally similar to interleukin 1.
强心甾类化合物哇巴因是膜Na⁺,K⁺-ATP酶的特异性抑制剂，已知其在低至10⁻⁷M的浓度下，就能抑制体外混合淋巴细胞培养中产生的或由各种刺激剂（PHA、Con A、PWM、可溶性抗原）诱导的人淋巴细胞增殖，而在此浓度下小鼠淋巴细胞增殖不受影响。哇巴因在转化的各个阶段均抑制大多数增殖反应参数。这一观察结果促使我们提出，哇巴因也可能通过抑制白细胞介素的产生起作用，已知在T细胞受刺激后的最初数小时，在有单核细胞存在的情况下会发生白细胞介素的产生。为了检验哇巴因对白细胞介素产生的可能影响，对在有或无抑制剂存在的情况下制备的受刺激人单核细胞的条件培养基，检测其促进小鼠胸腺细胞对PHA反应的能力。我们发现，即使在高浓度（2×10⁻⁶M）下，哇巴因非但没有产生预期的抑制作用，反而增强了刺激作用和/或小鼠胸腺细胞激活因子的产生。此外，未受刺激的人单核细胞在无血清培养中暴露于低浓度哇巴因（10⁻⁸至10⁻⁷M）24小时后的条件培养基，对小鼠胸腺细胞对PHA的反应显示出高度激活作用。经哇巴因处理后产生的这种可溶性因子由贴壁细胞产生，在功能上似乎与白细胞介素1相似。
