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目的明确模拟失重对白细胞介素2(IL-2)诱导的人自然杀伤(NK)细胞活性的影响。方法回转细胞模拟失重条件培养人NK细胞,CCK-8法检测NK细胞增殖活性、异硫氰酸荧光素标记的膜联素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)双标记结合流式细胞术检测细胞凋亡率、ELISA检测γ干扰素(IFN-γ)产生水平,反转录PCR检测NK细胞IL-12β1和IL-12β2受体mRNA水平。结果与正常重力组相比,模拟失重培养24 h和48 h后,IL-2诱导的NK细胞增殖水平分别下降13.6%和31%,凋亡率分别增加8%和19%,IFN-γ分泌水平分别降低25.2%和47.8%,杀伤活性分别下降7%和18%,IL-12刺激IL-2预处理NK细胞产生IFN-γ的水平分别降低21.8%和58.8%,同时,NK细胞IL-12β1和IL-12β2受体mRNA水平显著降低。结论模拟失重抑制IL-2诱导的NK细胞增殖、IFN-γ产生和杀伤活性,并进一步通过下调IL-12受体mRNA表达抑制IL-12诱导的IFN-γ产生。
Objective To determine the effect of simulated weightlessness on interleukin 2 (IL-2) -induced natural killer (NK) cell activity. Methods The human NK cells were cultured under simulated conditions of weightlessness by rotating cells. The proliferation activity of NK cells was detected by CCK-8 assay. The double labeled conjugate Ⅴ-FITC / PI labeled conjugate flow cytometry The rate of apoptosis was detected by cytometry. The production of IFN-γ was detected by ELISA. The levels of IL-12β1 and IL-12β2 mRNA were detected by reverse transcription polymerase chain reaction (RT-PCR). Results Compared with normal gravity group, the proliferation of NK cells induced by IL-2 decreased by 13.6% and 31%, respectively, and the apoptosis rate increased by 8% and 19%, respectively, at 24 h and 48 h after simulated weightlessness, IL-2 pretreatment IL-2 pretreatment reduced the levels of IFN-γ by 21.8% and 58.8%, respectively. In the meantime, the levels of IL-12 in NK cells decreased by 25.2% and 47.8%, and the cytotoxicity decreased by 7% and 18% 12β1 and IL-12β2 receptor mRNA levels were significantly reduced. Conclusion Simulated weight loss can inhibit IL-2-induced NK cell proliferation, IFN-γ production and cytotoxicity, and further inhibit IL-12-induced IFN-γ production by down-regulating IL-12 receptor mRNA expression.