米诺环素对小鼠T淋巴细胞体外活化和增殖的影响

研究米诺环素(MNC)对刀豆蛋白A(ConA)刺激的小鼠T细胞体外活化和增殖的影响,并对其免疫调节作用进行初步探讨。以ConA刺激小鼠淋巴结来源的淋巴细胞,以不同终浓度的MNC与T细胞共培养,荧光抗体染色结合流式细胞术,检测T细胞早期活化抗原CD69的表达;用羧基荧光素双醋酸盐琥珀酰(CFDA-SE)染色结合流式细胞术,并用ModFit软件分析T细胞增殖相关指数。结果:终浓度为10、25和50μmol/L的MNC显著抑制ConA诱导的T细胞CD69的表达,由ConA组的71.20%±1.08%,分别降低为64.43%±1.39%、53.34%±1.26%和33.65%±1.91%;CFDA-SE染色结果显示,培养48 h和72 h的ConA组T细胞的增殖指数(PI)分别为1.57±0.03和2.06±0.02,各浓度的MNC对ConA刺激的T细胞增殖具有明显的抑制作用,以50μmol/L的MNC抑制作用最明显,48 h和72 h的PI分别为1.01±0.02和1.03±0.01,与相应时间的对照组比较,均有统计学意义(P<0.01)。MNC能有效地抑制小鼠T细胞的体外活化和增殖。具有独立于其抗菌活性的抗炎作用。

Effect of minocycline on the in vitro activation and proliferation of murine T lymphocytes

In the present studiy,the effect of minocycline on the activation and proliferation of murine lymphocytes was investigated in vitro in order to elucidate its immunosuppressive mechanism,in which the T lymphocytes derived from lymph nodes of mice were stimulated with ConA and cocultured with different concentrations of minocycline.The expression of the early activated antigen CD69 on T lymphocytes was detected by fluoresent mono-antibodies and proliferation was detected by carboxyl fluorescein diacetate-succinimide ester(CFDA-SE) staining combined with flow cytometry,and the proliferation index(PI) was analyzed by means of ModFit software.It was demonstrated that the expression of CD69 on the ConA induced T lymphocytes was significantly inhibited by the final concentration of minicycline at 10,25 and 50 μmol/L,reducing from 71.20%±1.08% before treatment with minocycline to 64.43%±1.39%,53.34%±1.26% and 33.65%±1.91% after treatment with 10,25 and 50 μmol/L of minocycline respectively.As demonstrated by CFDA-SE staining,the proliferation induces of T lymphocytes at 48 hand 72 h in ConA induced group were 1.57±0.03 and 2.06±0.02.Minocyline at different concentration used all showed inhibitory effect on the ConA induced proliferation of T lymphocytes with the most significant effect on 50 μmol/L of minocycline,in which the PI at 48 h and 72 h of cultivation were 1.01±0.02 and 1.03±0.01 respectively.Significant difference existed in comparison with the control groups at the corresponding time of cultivation(P<0.01).It is concluded that minocycline can effectively inhibit the ConA induced activation and proliferation of murine T lymphocytes in vitro and it exhibits anti-inflammatory effect independent to its antimicrobial activity.