CAJ | 학술논문

目的观察模拟微重力条件下实验性自身免疫性脑脊髓炎(EAE)大鼠淋巴细胞细胞功能的变化.方法尾根免疫MBP68-86建立EAE模型;12d时处死大鼠,分离培养淋巴结细胞;利用旋转式细胞培养系统进行模拟微重力培养;不同时间点,增殖实验检测细胞增殖、流式检测细胞凋亡、T细胞亚型、Th细胞亚群变化;酶联免疫吸附试验(ELISA)检测上清中细胞因子的变化.结果 EAE淋巴细胞针对MBP_(68-86)抗原特异性增殖,在培养20h,60h,80h时,增殖受到抑制(t=3.859,P<0.01;t=5.933,P<0.001;t=7.613,P<0.001);40h时,增殖受到促进(t=4.015,P<0.01).培养24h时,微重力组坏死及凋亡细胞增高(t=3.998,P<0.01;t=3.705,P<0.01),Th细胞降低(t=4.111,P<0.01),TH1、TH17和调节性T细胞(Treg)亚群比例升高(t=2.743,P<0.05;t=4.362,P<0.01;t=2.945,P<0.05).微重力培养40h时,IFN-γ,TNF-α吨浓度明显升高(t=4.056,P<0.01;t=4.666,P<0.01),TGF-β、IL-6、IL-4、IL-17浓度明显降低(t=2.855,P<0.05;t=2.644,P<0.05;t=3.154,P<0.05;t=2.732,P<0.05).结论除40h外,MBP_(68-86)特异性淋巴细胞增殖在模拟微重力环境中被抑制;细胞死亡增多、Th细胞减少,对EAE起重要作用的细胞因子的浓度改变.
목적 관찰모의미중력조건하실험성자신면역성뇌척수염(EAE)대서림파세포세포공능적변화.방법 미근면역MBP68-86건립EAE모형;12d시처사대서,분리배양림파결세포;이용선전식세포배양계통진행모의미중력배양;불동시간점,증식실험검측세포증식、류식검측세포조망、T세포아형、Th세포아군변화;매련면역흡부시험(ELISA)검측상청중세포인자적변화.결과 EAE림파세포침대MBP_(68-86)항원특이성증식,재배양20h,60h,80h시,증식수도억제(t=3.859,P<0.01;t=5.933,P<0.001;t=7.613,P<0.001);40h시,증식수도촉진(t=4.015,P<0.01).배양24h시,미중력조배사급조망세포증고(t=3.998,P<0.01;t=3.705,P<0.01),Th세포강저(t=4.111,P<0.01),TH1、TH17화조절성T세포(Treg)아군비례승고(t=2.743,P<0.05;t=4.362,P<0.01;t=2.945,P<0.05).미중력배양40h시,IFN-γ,TNF-α둔농도명현승고(t=4.056,P<0.01;t=4.666,P<0.01),TGF-β、IL-6、IL-4、IL-17농도명현강저(t=2.855,P<0.05;t=2.644,P<0.05;t=3.154,P<0.05;t=2.732,P<0.05).결론 제40h외,MBP_(68-86)특이성림파세포증식재모의미중력배경중피억제;세포사망증다、Th세포감소,대EAE기중요작용적세포인자적농도개변.
Objective Culture auto immunogen reactive, hypersensitive lymphocytes in the model microgravity (MMG). Explore the effects of MMG on the function of objective lymphocytes. Methods Induced experimental autoimmune encephalomyelitis (EAE) by MBP_(68-86) injection in the tail base. 13 days later, rats were sacrificed and lymphocytes were isolated. MMG was implemented by rotating cell culture system ( RCCS) . Hours later, cell proliferation was estimated by CCK-8, cell apoptosis and subtypes of T cells and Th cells were analysised by flow cytometry, and the level of cytokines in the supernatant was examined by ELISA Kits. Results The primary lymphocytes were reactive to the specific antigen ( MBP_(68-86)). Cell proliferation was inhibited on 20h, 60h, 80h in MMG(t =3. 859,P<0.01; t=5. 933,P<0. 001; t =7. 613 ,P <0.001) , but increased on 40 h( t = 4.015, P < 0. 01). Cell apoptosis and necrosis were increased apparently (t = 3.998, P<0.01;t=3.705,P<0.01). The percentage of Th cell was decreased in MMG(t=4.111,P< 0.01). Thepercentage of Thl, Thl7 and Treg subsets were increased in MMG(t = 2. 743,P<0. 05; t =4. 362,P<0.01; t=2.945,P<0.05). The level of IFN-γ and TNF-α were increased (t = 4.056,P<0.01; t =4.666, P< 0.01) and the level of TGF-β IL-6, IL4 and IL-17 Were decreased on 40h MMG culture (t = 2.855, P<0.05; t=2.644,P<0.05; t =3.154,P<0.05; t =2.732,P<0.05). Conclusion The cell proliferation was suppressed in MMG except on 40 h. More lymphocytes were dead in MMG,and the percentage of Th cells were decreased. The subsets of Th cells were changed in MMG and the effects of MMG on the cytokines which were important in the pathogenesis of EAE were various.