青蒿琥酯逆转人卵巢癌3AO细胞所致T淋巴细胞的免疫抑制研究

Artesunate for reversing human ovarian cancer 3AO cell caused T lymphocytes immunosuppression

目的研究新型抗瘤中药制剂青蒿琥酯(ART)对T淋巴细胞表面免疫因子的影响。方法噻唑蓝(MTT)法筛选对卵巢癌3AO细胞增殖无明显影响的最高ART浓度。收集经ART预处理后的3AO细胞培养上清液(ART-S组)及连续两次再培养上清液(ART-S1组和ART-S2组),以不经ART作用的相应同步培养的3AO细胞培养上清液作对照(Con-S组、Con-S1组、Con-S2组),以完全培养基代替3AO细胞培养上清液组为正常对照组(Con组),流式细胞计数法分析各组上清液对T细胞表面CD3ε+、CD3ε+ζ+、IL-2Rα+免疫功能指标的影响。结果对3AO细胞增殖无明显影响的最高ART浓度为12.50 mg/L。Con组的CD3ε+、CD3ε+ζ+、IL-2Rα+细胞表达阳性率分别为(86.07±10.05)%、(25.09±5.84)%、(39.11±5.83)%。Con-S组3项免疫指标明显受抑制,抑制率分别为(50.17±8.02)%、(87.51±9.17)%、(64.84±9.74)%。ART直接作用后可明显下调单纯的3AO肿瘤细胞所致的3项T细胞免疫功能抑制。结论青蒿琥酯对人卵巢癌3AO细胞所致T淋巴细胞免疫功能有一定的逆转作用。

Objective To investigate the effect of new type anti-tumor Chines medicine artesunate（ART）on T lymphocyte surface immune factors by in vitro experiment.Methods The MTT method was used to screen the maximum ART concentration without significant effect on human ovary caner 3 AO cells proliferation.The 3 AO cell culture supernatant and successive two times re-culture supernatants after ART treatment were collected as the ART-S group,ART-S1 group and ART-S2 group,and corresponding synchronized 3 AO cell culture supernatant without ART action served as the control groups（Con-S group,Con-S1 group and Con-S2 group）,the complete medium instituting the 3 AO cell culture supernatant served as the normal control group（Con group）.The effects of supernatant in each group on T lymphocyte surface immune functional indexes of CD3ε^＋,CD3ε^＋ζ^＋and IL-2 Rα^＋were analyzed by using flow-cytometry.Results The maximum ART concentration without significant effect on 3 AO cell proliferation was 12.50 mg/L.The expression positive rates of CD3ε^＋,CD3ε^＋ζ^＋and IL-2 Rα＋cells in the Con group were（86.07±10.05）%,（25.09±5.84）% and（39.11±5.83）%respectively.Those indexes in the Con-S group were significantly repressed（P〈0.05）,the suppression rates were（50.17±8.02）%,（87.51±9.17）% and（64.84±9.74）% respectively.After ART direct action,the suppression of 3 Tlymphocyte immune function caused by simply3 AO tumor cell was obviously down-regulated.Conclusion ART has a certain reversion effect on T lymphocyte immune function caused by human ovarian cancer 3 AO cells.