中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
5期
794-797
,共4页
徐丽%俞建东%吕丽虹%龙天柱%黄永亨%闵军%万云乐
徐麗%俞建東%呂麗虹%龍天柱%黃永亨%閔軍%萬雲樂
서려%유건동%려려홍%룡천주%황영형%민군%만운악
大蒜素%酸性微环境%自然杀伤细胞
大蒜素%痠性微環境%自然殺傷細胞
대산소%산성미배경%자연살상세포
Allicin%Acidic microenvironment%NK cells
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pH 5.6、pH 6.5和pH 7.2的完全RPMI 1640培养基对大鼠脾脏CD3-NKR-P1+NK细胞进行悬浮培养,并给予30 mg/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)-γ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pH 7.2、pH 6.5和pH 5.6三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pH 5.6、培养16 h时达最高33.3%.与之相对应,NK细胞分泌的IFN-γ达(64.59±0.09)ng/L,较培养4 h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.
目的 觀察體外痠性培養環境下大蒜素對自然殺傷(NK)細胞功能活性的影響,併探討其機製.方法 以pH 5.6、pH 6.5和pH 7.2的完全RPMI 1640培養基對大鼠脾髒CD3-NKR-P1+NK細胞進行懸浮培養,併給予30 mg/L濃度的大蒜素進行處理.酶聯免疫吸附試驗(ELISA)檢測細胞培養液中榦擾素(IFN)-γ的分泌水平,流式細胞儀檢測NK細胞的增殖和凋亡率,乳痠脫氫酶法檢測NK細胞的功能活性.結果 痠性培養環境下大鼠脾髒NK細胞的增殖能力明顯下降,NK細胞功能活性明顯受阻.在pH 7.2、pH 6.5和pH 5.6三種不同的培養條件下,大蒜素對NK細胞增殖率的影響錶現為隨培養環境的pH降低反而升高,以pH 5.6、培養16 h時達最高33.3%.與之相對應,NK細胞分泌的IFN-γ達(64.59±0.09)ng/L,較培養4 h時升高28%,且對小鼠淋巴瘤Yac-1細胞的殺傷活性也達到最高水平.結論 大蒜素可通過提高NK細胞IFN-γ分泌水平明顯改善痠性培養環境下NK細胞的功能活性.
목적 관찰체외산성배양배경하대산소대자연살상(NK)세포공능활성적영향,병탐토기궤제.방법 이pH 5.6、pH 6.5화pH 7.2적완전RPMI 1640배양기대대서비장CD3-NKR-P1+NK세포진행현부배양,병급여30 mg/L농도적대산소진행처리.매련면역흡부시험(ELISA)검측세포배양액중간우소(IFN)-γ적분비수평,류식세포의검측NK세포적증식화조망솔,유산탈경매법검측NK세포적공능활성.결과 산성배양배경하대서비장NK세포적증식능력명현하강,NK세포공능활성명현수조.재pH 7.2、pH 6.5화pH 5.6삼충불동적배양조건하,대산소대NK세포증식솔적영향표현위수배양배경적pH강저반이승고,이pH 5.6、배양16 h시체최고33.3%.여지상대응,NK세포분비적IFN-γ체(64.59±0.09)ng/L,교배양4 h시승고28%,차대소서림파류Yac-1세포적살상활성야체도최고수평.결론 대산소가통과제고NK세포IFN-γ분비수평명현개선산성배양배경하NK세포적공능활성.
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 cytometry. 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 supernatant 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 cells was also found strongest under the condition of pH 5. 6 cultured for 16 h. Conclusion Allicin favored to enhance the cytotoxicity of NK cells under the acidic cultured condition, which might be related to the increase of IFN-γ production.
