酸性微环境下大蒜素增强大鼠自然杀伤细胞的功能活性  

Enhancement of rat nature killer cell activity by allicinin vitrounder acidic microenvironment

作　　者：徐丽[1] 俞建东[2] 吕丽虹[2] 龙天柱[2,3] 黄永亨[2] 闵军[2] 万云乐[2] 

机构地区：[1]广东轻工职业技术学院食品与生物工程系,广州510300 [2]中山大学孙逸仙纪念医院肝胆胰外科 [3]广州市妇女儿童医院普外科

出　　处：《中华实验外科杂志》2011年第5期794-797,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30671987）

摘　　要：目的观察体外酸性培养环境下大蒜素对自然杀伤（NK）细胞功能活性的影响，并探讨其机制。方法以pH5．6、pH6．5和pH7．2的完全RPMI1640培养基对大鼠脾脏CD3-NKR—P1＋NK细胞进行悬浮培养，并给予30mg／L浓度的大蒜素进行处理。酶联免疫吸附试验（ELISA）检测细胞培养液中干扰素（IFN）-1的分泌水平，流式细胞仪检测NK细胞的增殖和凋亡率，乳酸脱氢酶法检测NK细胞的功能活性。结果酸性培养环境下大鼠脾脏NK细胞的增殖能力明显下降，NK细胞功能活性明显受阻。在pH7．2、pH6．5和pH5．6三种不同的培养条件下，大蒜素对NK细胞增殖率的影响表现为随培养环境的pH降低反而升高，以pH5．6、培养16h时达最高33．3％。与之相对应，NK细胞分泌的IFN-1达（64．59±0．09）ng／L，较培养4h时升高28％，且对小鼠淋巴瘤Yac-1细胞的杀伤活性也达到最高水平。结论大蒜素可通过提高NK细胞IFN-γ分泌水平明显改善酸性培养环境下NK细胞的功能活性。Objective To investigate the role of allicin on rat nature killer （NK） cell activity in vitro under acidic microenvironment, and its possible mechanism. Methods CD3 - NKR-P1 ＋ NK cells isolated from the rat spleen were cultured in the complete RPMI 1640 medium （ pH 5.6, pH 6. 5, or pH 7.2 respectively）, and treated with allicin at final concentration of 30 mg/L. Enzyme linked immunosorbent assay （ELISA） was used to determine supernatant interferon （IFN）-γ levels. The percentage of NK cells proliferation and apopotosis was analyzed by flow eytometry. NK cell cytotoxicity toward YAC-1 tumor cells was detected by LDH release assay. Results Proliferation and cytotoxicity of NK cells were significantly suppressed by acidic microenvironment in vitro. Under the cultured condition of acidic pH below 7. 2, allicin seemed to promote NK cells proliferation, which reached to highest level of 33% at pH 5.6 cultured for 16 h. Correspondingly, at pH of 5.6, allicin induced a marked increase of IFN-γ concentration in the sapernatant from （50. 07± 0. 38） （ cultured for 4 h） to （ 64. 59 ＋ 0. 09） ng/L （ cultured for 16 h）. The cytotoxicity of NK cells toward YAC-1 tumor ceils was also found strongest under the condition of pIff 5. 6 cultured for 16 h. Conclusion Allicin favored to enhance the cytotoxicity of NK ceils under the acidic cultured condition, which might be related to the increase of IFN-3, production.

关 键 词：大蒜素 酸性微环境 自然杀伤细胞 

分 类 号：R285.5[医药卫生—中药学]