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目的研究ω-3多不饱和脂肪酸对树突状细胞的影响。方法采用GM—CSF及白细胞介素(IL)-4诱导人外周单核细胞获得非成熟树突状细胞(DCs),然后以不饱和脂肪酸二十碳五烯酸(EPA),二十二碳六烯酸(DHA)或饱和脂肪酸硬脂酸(SA)处理细胞24 h,并将其分为5组:非成熟DCs组、成熟DCs组、EPA处理组、DHA处理组和SA处理组。内毒素脂多糖(LPS)诱导细胞成熟后用流式细胞仪分别检测DCs表面抗原提呈分子HLA-DR及共刺激分子CD40、CD80、CD86 的表达率;采用酶联免疫吸附试验(ELISA)法检测各组细胞上清中白细胞介素-12及肿瘤坏死因子 (TNF)-α的含量:混合T淋巴细胞反应检测DCs刺激细胞增殖能力。结果非成熟DCs组CD80、 CD86及HLA-DR的表达率为(48．7±17．5)％、(57．3±16．9)％及(78．5±15．1)％,成熟DCs组这 3种分子高表达,其表达率为分别为(86．3±10．8)％、(77．4±13．5)％及(79．6±14．5)％,EPA及 DHA处理组CD80、CD86及HLA-DR的表达率分别为(65．5±17．2)％、(60．2±9．1)％、(56．9± 12．3)％和(64.7±13．5)％、(57．7±13．6)％、(64．9±5．8)％,与成熟DCs组相比有统计学意义 (P<0．05),而SA处理的DCs表达率分别为(88．1±4．5)％、(71．2±14．5)％、(75．9±10．7)％与成熟组相比差异无统计学意义(P>0．05)。细胞上清IL—12及TNF-α的含量,5个组分别为(70．8± 1．9)、(286．8±80．4)、(158．5±43．5)、(161．0±20．1)、(271．5±45．6)和(4．7±2．3)、(32．1± 12．5)、(13．7±4．8)、(9．0±6．5)、(27．6±11．4)ng／L,EPA及DHA处理组与成熟DCs组相比差异有统计学意义(P<0．01),而SA组与成熟DCs组相比差异无统计学意义(P>0．05)。EPA及DHA 组其DCs刺激T细胞增殖的能力亦显著下降(P<0．05),并且随着EPA及DHA浓度的增加,其下降越明显。结论ω-3多不饱和脂肪酸可以在体外抑制树突状细胞免疫表型的表达及细胞因子的释放,降低其刺激T细胞增殖的能力。
Objective To investigate the effect of ω-3 polyunsaturated fatty acids on dendritic cells. Methods Human peripheral blood mononuclear cells were induced by GM-CSF and interleukin-4 (IL-4) to obtain immature dendritic cells (DCs). The dendritic cells were treated with unsaturated fatty acids eicosapentaenoic acid (EPA) The cells were treated with DHA or SA for 24 h and divided into 5 groups: immature DCs group, mature DCs group, EPA-treated group, DHA-treated group and SA-treated group. The expression of HLA-DR and co-stimulatory molecules CD40, CD80 and CD86 on DCs were detected by flow cytometry after induced by endotoxin lipopolysaccharide (LPS). ELISA and ELISA The levels of interleukin-12 and tumor necrosis factor-α (TNF-α) in the supernatant of each group were detected: mixed T lymphocyte reaction was used to detect the ability of DCs to stimulate cell proliferation. Results The expression rates of CD80, CD86 and HLA-DR in non-mature DCs group were (48.7 ± 17.5)%, (57.3 ± 16.9)% and (78.5 ± 15.1)%, respectively (86.3 ± 10.8)%, (77.4 ± 13.5)% and (79.6 ± 14.5)%, respectively. The rates of EPA and The expression rates of CD80, CD86 and HLA-DR in DHA group were (65.5 ± 17.2)%, (60.2 ± 9.1)%, (56.9 ± 12.3)% and ± 13.5%, 57.7 ± 13.6% and 64.9 ± 5.8%, respectively, which were significantly higher than those in mature DCs group (P <0.05) (88.1 ± 4.5)%, (71.2 ± 14.5)% and (75.9 ± 10.7)%, respectively, had no significant difference compared with the mature group (P > 0.05). The levels of IL-12 and TNF-α in the supernatants were (70.8 ± 1.9), (286.8 ± 80.4), (158.5 ± 43.5) and (161 .0 ± 20.1), (271.5 ± 45.6) and (4.7 ± 2.3), (32.1 ± 12.5), (13.7 ± 4.8), (9 .0 ± 6.5) and (27.6 ± 11.4) ng / L, respectively. There was a significant difference between EPA and DHA groups and mature DCs group (P <0.01) There was no significant difference between DCs group (P> 0.05). The ability of DCs to stimulate T cell proliferation in the EPA and DHA groups also decreased significantly (P <0.05), and the more significant the decline was with the increase of EPA and DHA concentrations. CONCLUSION: Omega-3 polyunsaturated fatty acids can inhibit the immunophenotypic expression of dendritic cells and the release of cytokines in vitro, reducing their ability to stimulate T cell proliferation.