棉酚对多克隆激活剂活化人T淋巴细胞的抑制作用

目的:以体外培养的人外周血单个核细胞为材料,研究多酚类抗生育药物棉酚对正常人外周血T淋巴细胞体外活化作用的影响。方法:利用双荧光染色流式细胞术分析棉酚对人T淋巴细胞在丝裂原或佛波醇酯刺激下早期活化抗原CD69表达的影响。结果:100μmol/L的棉酚预先与T细胞温育,能完全抑制10mg/L植物血凝素(PHA)刺激的CD3^＋T细胞活化抗原CD69的表达,又可阻断10^－7mol/L佛波醇酯(PDB)对T细胞的活化作用。这种抑制作用呈剂量依赖性,棉酚对PDB及PHA抑制作用的IC₅₀分别为(35.7±2.9)μmol/L和(32.8±1.5)μmol/L。此外,棉酚对CD3^－的淋巴细胞CD69表达也有类似的抑制作用。然而,它对多克隆激活剂引起的T细胞表面分子CD3的下调无明显影响。结论:在体外活化模型中,棉酚能够同时抑制多克隆激活剂PHA和PDB对T细胞的活化作用,提示其作用部位可能位于PKC或下游,并提示棉酚具有潜在的免疫调节作用。

Inhibitory effects of gossypol on the activation of human T-lymphocytes stimulated with polyclonal activators

AIM: Peripheral blood mononuclear cells (PBMC) were cultured in vitro to study the effect of gossypol, a polyphenolic antifertility agent, on the activation of normal human T cells. METHODS: Double fluorescent staining together with flow cytometry was adopted to analyze the influence of gossypol on expression of the early activation antigen CD69 on T-lymphocytes under stimulation of mitogen or phorbol ester. RESULTS: Analysis of T cell activation in vitro revealed that preincubation of PBMC with 100 μmol/L gossypol could completely inhibit the expression of early activation marker CD69 on CD3+ T cells in response to 10　mg/L PHA, and block T cell activation by 10-7 mol/L PDB as well. The suppression of CD69 expression was dose-dependent and IC50 of gossypol on PDB and PHA were (35.7±2.9) μmol/L and (32.8±1.5) μmol/L(眘), respectively. Besides, gossypol had similar inhibitory effect on CD69 expression of CD3- lymphocytes. However, it did not have any significant effect on T cell surface molecule CD3 down-regulation. CONCLUSION: Gossypol could inhibit T cell activation in vitro in response to polyclonal activators, both PHA and PDB, suggesting that its action site may be at PKC or its downstream and that gossypol possessed potential immuno-regulatory effect.