失代偿期乙肝肝硬化患者的骨髓间充质干细胞的免疫调节作用

目的 观察失代偿期乙型肝炎肝硬化患者的骨髓间充质干细胞（BMSCs）对自体外周血淋巴细胞增殖及调节性T细胞（Tregs）亚群的影响.方法 从8例失代偿性肝硬化患者骨髓中分离培养MSCs,采用流式细胞仪分析鉴定MSCs的纯度,分离患者外周血淋巴细胞行羧基荧光素乙酰乙酸琥珀酰亚胺酯（CFDA SE）荧光染色,在植物血凝素（PHA）刺激下,培养基为自体血清,自体外周血淋巴细胞与BMSCs共培养,流式细胞术检测各组淋巴细胞增殖情况及CD4＋ CD25＋ CD127-Tregs频数.结果 流式细胞术显示：接触培养组及非接触培养组的细胞增殖较阳性对照组明显减低（P均＜0.01）,但亦显著高于阴性对照组（P均＜0.01）,其CD4＋ CD25＋ CD127-Tregs频率显著高于阳性及阴性对照组（P均＜0.01）;接触培养组与非接触培养组比较,淋巴细胞增殖及CD4＋ CD25＋ CD127-Tregs频数差异均无统计学意义（P均＞0.05）.结论 肝硬化患者BMSCs可抑制自体外周血淋巴细胞增殖、上调CD4＋ CD25＋ CD127-Tregs表达.

Immune regulatory effect of bone marrow mesenchymal stem cells of patient with hepatitis B virus related decompensated cirrhosis

Objective To study the effect of bone marrow mesenchymal stem cells （BMSCs） on autologous peripheral blood lymphocyte proliferation and regulatory T cells （Tregs）subsets in vitro in patients with hepatitis B virus related decompensated cirrhosis.Methods MSCs were isolated and expanded from human bone marrow blood of eight patients with decompensated cirrhosis.The purity of MSCs was identified by flow cytometry.lymphocytes were isolated from the peripheral blood of patients and stained with carboxy fluoresce indiacetate succinimidyl ester （CFDA SE）.The BMSCs and peripheral blood lymphocytes from patients were added into wells containing autologous serum in the presence of phytohemagglutinin （PHA）.lymphocytes proliferation rate and CD4 ＋ CD25 ＋ CD127-Tregs frequency were detected by flow cytometry.Results Flowcytometric analysis showed that lymphocyte proliferation in contact co-culture group and noncontact co-culture group were much lower than positive control （all P 〈 0.01）,and significantly higher than negative control （all P 〈 0.01）,while CD4＋ CD25＋ CD127-Tregs in contact co-culture group and noncontact co-culture group were much higher than positive control and negative control （all P 〈 0.01）.Besides,there was no significant difference on lymphocyte proliferation rate or Tregs frequency between contact co-culture group and non-contact co-culture group（all P 〉 0.05）.Conclusion BMSCs can inhibit proliferation of autologous peripheral blood lymphocytes and increase expression of CD4 ＋ CD25 ＋ CD127-Tregsin patients with decompensated cirrhosis.