小鼠骨髓间充质干细胞对植物血凝素诱导小鼠脾细胞增殖后趋化因子受体表达的影响

Effects of mouse bone marrow mesenchymal stem cells on the expression of chemokine receptors of spleenocytes after phytohemagglutinin stimulation

目的观察小鼠骨髓间充质干细胞(mesenchymal stem cells,MSCs)在体外对小鼠脾细胞表达趋化因子受体的影响。方法用密度梯度离心法从小鼠骨髓中分离出小鼠骨髓间充质干细胞,经低糖DMEM培养基培养扩增。C57BL/6小鼠脾细胞以1×106/孔的密度接种于24孔板,加入植物血凝素(PHA),培养72h。实验分3组:A组按10%比例加入MSC;B组按1%比例加入MSC;对照组不加MSC。3d后收集悬浮的脾细胞进行流式细胞术检测小鼠脾细胞趋化因子受体CXCR3,CCR5,CCR7表达的变化。结果CD3+CCR5+、CD3+CCR7+在A,B组及对照组中表达的差异均有统计学意义(均P<0.01);以A组表达率最高,B组次之,对照组最低;CD3+CXCR3+细胞在A组中表达较B组、对照组高(P<0.05),B组和对照组之间的表达差异无统计学意义。结论骨髓间充质干细胞在一定浓度下对小鼠脾细胞增殖后趋化因子受体CXCR3,CCR5,CCR7表达有上调作用。

Objective To explore the effect of mouse bone marrow mesenchymal stem cells （MSCs） on the expression of chemokine receptors in T lymphocytes in vitro. Methods Mouse bone marrow MSCs were separated with Percoll, cultured and expanded in low glucose DMEM. C57BL/6 mouse slpeenocytes were cultured in the 24-hole flasks by the density of 1×10^6/hole. Phytohemagglutinin （ PHA ） was then added to the holes and cultured for 72 hrs. This study consisted of three groups. Groups A and B were co-cultured by adding MSCs as the ratio of 0.1 and 0.01 to slpeenocytes respectively. The control group was cultured without MSCs. Three days later the suspended spleenocytes were harvested for detecting the expression of three chemokine receptors CXCR3, CCR5 and CCR7 in T lymphocytes by the flow cytometry. Results The expression of CD3^＋ CCR5^＋ and CD3^＋ CCR7^＋ were statistically different among the three groups. Group A had the strongest expression, followed by group B and the control group. The expression of CD3^＋ CXCR3^＋ in group A was statistically higher than that in group B and the control group. Conclusions MSCs could up-regulate the expression of chemokine receptors CXCR3, CCR5 and CCR7 in T lymphocytes stimulated by PHA.