CAJ | 학술논문

目的：探讨骨髓间充质干细胞（MSCs）对单向混合淋巴细胞反应（MLR）培养体系中被激活的T淋巴细胞增殖和miRNA-155表达的影响。方法体外培养SD大鼠MSCs ，采用流式细胞仪检测MSCs细胞表面标记CD11b/c、CD34、CD44和CD90；分别通过成骨和成脂诱导鉴定MSCs的分化潜能。免疫磁珠分选SD大鼠脾T淋巴细胞，并行纯度及活性检测。以SD大鼠T淋巴细胞作为反应细胞、丝裂霉素C灭活的Wistar大鼠单个核细胞作为刺激细胞建立单向MLR培养体系。CCK-8法检测MSCs对MLR中被激活的 T淋巴细胞增殖的影响。Rea1-time PCR法检测MSCs对MLR中被激活的T淋巴细胞miRNA-155表达的影响。结果流式细胞仪鉴定结果显示：CD44及CD90的阳性细胞数分别为（98．9％±0．8％）、（98．1％±0．9％），而CD11b/c及CD34的阴性细胞数分别为（85．1％±0．6％）、（98．0％±0．8％）。培养的MSCs具有成骨和成脂分化潜能。所分选的T淋巴细胞纯度为（91．9％±1．2％）。MSCs可明显抑制单向MLR培养体系中激活的 T淋巴细胞的增殖和miRNA-155表达，且这种抑制作用呈明显的浓度依赖性。结论 MSCs能够降低被激活的 T 淋巴细胞中miRNA-155的表达，这可能在MSCs调节T淋巴细胞增殖和免疫功能的过程中发挥了重要作用。
목적：탐토골수간충질간세포（MSCs）대단향혼합림파세포반응（MLR）배양체계중피격활적T림파세포증식화miRNA-155표체적영향。방법체외배양SD대서MSCs ，채용류식세포의검측MSCs세포표면표기CD11b/c、CD34、CD44화CD90；분별통과성골화성지유도감정MSCs적분화잠능。면역자주분선SD대서비T림파세포，병행순도급활성검측。이SD대서T림파세포작위반응세포、사렬매소C멸활적Wistar대서단개핵세포작위자격세포건립단향MLR배양체계。CCK-8법검측MSCs대MLR중피격활적 T림파세포증식적영향。Rea1-time PCR법검측MSCs대MLR중피격활적T림파세포miRNA-155표체적영향。결과류식세포의감정결과현시：CD44급CD90적양성세포수분별위（98．9％±0．8％）、（98．1％±0．9％），이CD11b/c급CD34적음성세포수분별위（85．1％±0．6％）、（98．0％±0．8％）。배양적MSCs구유성골화성지분화잠능。소분선적T림파세포순도위（91．9％±1．2％）。MSCs가명현억제단향MLR배양체계중격활적 T림파세포적증식화miRNA-155표체，차저충억제작용정명현적농도의뢰성。결론 MSCs능구강저피격활적 T 림파세포중miRNA-155적표체，저가능재MSCs조절T림파세포증식화면역공능적과정중발휘료중요작용。
Objective To explore the effect of bone mesenchymal stem cells (MSCs) on cell pro-liferation and miRNA-155 expression of activated T lymphocytes in the one-way mixed lymphocyte reaction (MLR) system . Methods MSCs were isolated from SD rats and cultured in vitro . Cells were identified with a flow cytometer for the expression of CD11b/c ,CD34 ,CD44 and CD90 . The differentiation ability of MSCs was identified by osteogenic and adipogenic induction . The one-way MLR system was estab-lished with T lymphocytes isolated from the spleens of SD rats by immunomagnetic beads and inactivated mononuclear cells solated from the spleens of Wistar rats . The effect of MSCs on cell proliferation and miRNA-155 expression of activated T lymphocytes in the one-way MLR system were respectively detected by CCK-8 assay and rea1-time PCR . Results Cultured MSCs were negative for CD11b/c and CD34 [(85 .1% ± 0 .6% ) and (98 .0% ± 0 .8% ) , respectively ] but positive for CD44 and CD90 [(98 .9%± 0 .8% ) and (98 .1% ± 0 .9% ) ,respectively] . The cells had the ability of osteogenic and adipogenic dif-ferentiation . The proliferation and miRNA-155 expression of activated T lymphocytes in the one-way MLR could be significantly inhibited by MSCs in a dose-dependent manner . Conclusions The expression of miRNA-155 in activated T lymphocytes can be down-regulated by MSCs ,which may play an important role in the regulaton of MSCs on cell proliferation and immune function of T lymphocytes .