CAJ | 학술논문

目的：体外实验研究青蒿琥酯（ ART）对人喉癌细胞所致T细胞免疫抑制的影响。方法制备经ART预处理的人喉癌Hep-2细胞培养上清（ART-S），收集连续2次再培养的上清（分别称为ART-S1、ART-S2），以不经ART作用的Hep-2及其上清作对照；分析对人外周血单个核细胞经光镜观察、噻唑蓝（ MTT）法测定的植物血凝素（ PHA）诱导淋巴细胞转化增殖反应，以及流式细胞计数（FCM）法检测的T淋巴细胞表面CD3ε＋、CD3ε＋ζ＋、IL-2Rα＋表达的影响。结果 Hep-2对5项免疫指标的抑制率分别为（52？．58±6．13）％、（50．34±7．26）％、（45．28±5．38）％、（39．74±5．27）％、（64．25±9．37）％。 ART作用后，可显著下调Hep-2所致T细胞免疫抑制（ P ＜0．01）；第1次再培养后，对淋巴细胞转化、诱导增殖、IL-2Rα表达抑制的逆转均明显降低，与ART-S比较差异有统计学意义（ P ＜0．01），对CD3ε＋及CD3ε＋ζ＋表达抑制的逆转持续增强，与ART-S比较差异有统计学意义（ P ＜0．05）；第2次再培养后，对淋巴细胞转化、诱导增殖、IL-2Rα表达抑制的逆转继续保持，与ART-S1比较差异无统计学意义（ P＞0．05），对CD3ε＋、CD3ε＋ζ＋表达抑制的逆转明显下降回复至 ART-S水平，差异无统计学意义（ P ＞0．05）。结论人喉癌Hep-2细胞可产生T细胞免疫抑制作用，ART可对其产生稳定的逆转作用。下调肿瘤免疫抑制是ART的重要抗瘤机制之一。
목적：체외실험연구청호호지（ ART）대인후암세포소치T세포면역억제적영향。방법제비경ART예처리적인후암Hep-2세포배양상청（ART-S），수집련속2차재배양적상청（분별칭위ART-S1、ART-S2），이불경ART작용적Hep-2급기상청작대조；분석대인외주혈단개핵세포경광경관찰、새서람（ MTT）법측정적식물혈응소（ PHA）유도림파세포전화증식반응，이급류식세포계수（FCM）법검측적T림파세포표면CD3ε＋、CD3ε＋ζ＋、IL-2Rα＋표체적영향。결과 Hep-2대5항면역지표적억제솔분별위（52？．58±6．13）％、（50．34±7．26）％、（45．28±5．38）％、（39．74±5．27）％、（64．25±9．37）％。 ART작용후，가현저하조Hep-2소치T세포면역억제（ P ＜0．01）；제1차재배양후，대림파세포전화、유도증식、IL-2Rα표체억제적역전균명현강저，여ART-S비교차이유통계학의의（ P ＜0．01），대CD3ε＋급CD3ε＋ζ＋표체억제적역전지속증강，여ART-S비교차이유통계학의의（ P ＜0．05）；제2차재배양후，대림파세포전화、유도증식、IL-2Rα표체억제적역전계속보지，여ART-S1비교차이무통계학의의（ P＞0．05），대CD3ε＋、CD3ε＋ζ＋표체억제적역전명현하강회복지 ART-S수평，차이무통계학의의（ P ＞0．05）。결론인후암Hep-2세포가산생T세포면역억제작용，ART가대기산생은정적역전작용。하조종류면역억제시ART적중요항류궤제지일。
Objective To investigate the effects of Artesunate ( ART ) on T lymphocytes ’ immunosuppression in human laryngocarcinoma cells in vitro .Met hods The supernatants from Hep-2 ( human laryngocarcinoma cell line ) were pretreated with ART ( ART-S) ,then the supernatants cultured for the first time and the second time were collected ( ART-S1 and ART-S2,respectively),and the corresponding supernatants and Hep-2 without ART treatment were served as control group.The effects of different supernatants on immune parameters in human peripheral blood mononuclear cells were analyzed , including the lymphocytes proliferation induced by phytohemagglutinin (PHA),which was detected by MTT and microscope observation,and expressions of CD3ε+,CD3ε+ζ+and IL-2Rα+were measured by flow cytometry (FCM).Results The inhibitory rates of five immune parameters caused by Hep -2 cells were52.58%±6.13%,50.34%±7.26%,45.28%± 5.38%,39.74%±5.27%,64.25%±9.37%,respectively.After pretreated with ART,the inhibition effects caused by Hep-2 were significantly decreased ( P <0.01 ).After the first culture, the reversing effect on the inhibition of lymphocytes transformation,induced proliferation and the expression of IL-2Rα+were significantly decreased in ART-S1 group, as compared with those in ART-S group ( P <0.01),and the reversing effects on the inhibition of CD3ε+and CD3ε+ζ+expressions were obviously kept,as compared with those in ART-S group ( P <0.05).After the second culture,there were no significant differences in , reversing effects on the inhibition of lymphocytes transformation , induced proliferation and the expression of IL-2Rα+between ART-S2 group and ART-S1 group ( P >0.05),nor were the reversing effects on the inhibition of CD3ε+and CD3ε+ζ+expressions between ART-S2 group and ART-S1 group ( P >0.05).Conclusion Human laryngocarcinoma Hep-2 cells have T lymphocytes immunosuppression effects .However Artesunate can steadily reversethe immunosuppression effects.Down-regulating tumor immunosuppression effect may be one of important anti -tumor actionmechanisms of Artesunate.