还原型谷胱甘肽抗白血病免疫佐剂作用实验研究

Effect of Reduced Glutathione as Anti-leukemic Immune Adjuvant

本研究探讨还原型谷胱甘肽(GSH)逆转反应性氧代谢物(ROM)对NK细胞抗白血病效应的抑制作用。在K562细胞、NK细胞的混合培养体系中分别加入富含单核细胞的单个核细胞(Mo)和白细胞介素-2(IL-2),观察ROM产量和K562细胞抑制率,然后分别加入GSH或二氢氯化物组胺(组胺),观察ROM产量及K562细胞抑制率的变化。结果表明:加入IL-2后,ROM的产量从33.17±25.02U/L增至223.59±59.41U/L(P<0.01),K562细胞抑制率从65.56%升至85.89%(P<0.01);在加入E/Mo=10/1、10/5、10/103种浓度的Mo后,ROM产量分别为389.79±43.83U/L,456.74±42.77U/L,601.42±21.92U/L,K562细胞抑制率分别为82.36%,81.36%,48.09%,加入组胺或GSH后,E/Mo=10/2时,ROM产量从389.79±43.83U/L,分别减至50.21±22.4U/L和-3.58±9.49U/L(P<0.05),随着GSH或组胺浓度的增加ROM产量逐减少,K562细胞抑制率从82.53%分别升至94.64%和96·39(P<0·05),ROM产量与K562细胞抑制呈负相关(P<0.05);E/Mo=10/5或10/10时,高浓度的GSH或组胺可使ROM产量减少,但K562细胞抑制率提高不明显(P>0.05)。结论:当E/Mo=10/2时,GSH逆转ROM强于组胺,提高NK细胞对K562的抑制率与组胺相似,但毒副作用轻微,GSH可能成为更理想的抗白血病的免疫佐剂。

To investigate the reversal effect of reduced glutathione（GSH） on suppression of NK cells by reactive oxygen metabofites （ROM） in K562 cells, interleukin-2 （IL-2） or mononuclear cell （Mo） was added in cultured cell line of K562 cells and NK cells, the yield of ROM and K562 cell suppression rate were observed. Then the histamine dihydrochloride （DHT） or GSH was added in the mixed cultured cell lines, the ROM production and K562 cell suppression rate were observed. The results showed that the ROM yield increased from 33.17 ±5.08 U/L to 223.59 ±9.41 U/L by IL-2, and K562 cell suppression rate increased from 65.56% to 85.89% by IL-2 （P 〈0.01 ）. The ROM yields were 389.79 ± 43.83 U/ml, 456.74 ± 42.77 U/ml and 601.42 ± 21.92 U/ml respectively, and K562 cell suppression rates were 82.36%, 81.36% and 48.09% respectively, when Mo was added in the mixed cultured cell lines under ratios of E/Mo being 10/2, 10/5 and 10/10. When E/Mo was 10/2, DHT or GSH was added in the mixed cultured cell line ROM yield decreased from 389.79 ± 3.83 U/L to 50.21 ± 2.4 U/L or - 3.58 ± 9.49 U/L （ P 〈 0.05 ） respectively. With increase of concentration of DHT or GSH, the ROM yield in the mixed cultured cell line decreased （P 〈0. 05）, the K562 cell suppression rate increased from 82.53% to 94.64% or 96.39% （ P 〈 0.05 ）, the more ROM yield, the less K562 suppression rate （P〈0.05）. When E/Mo is 10/5 or 10/10, the ROM yield decreased by the high concentration of DHT or GSH （ P 〈 0. 05 ）, but the K562 cell suppression rate not increased by every concentration of DHT or GSH. GSH was as effective as DHT in the reversing ROM and increasing K562 cell suppression rate. It is concluded that GSH may reverse ROM and increase K562 cell suppression rate, and GSH is as effective as DHT, but GSH has less side-effect than DHT. Therefore, GSH would be better antileukemia immune adjuvant.