CAJ | 학술논문

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.
목적 관찰실대상기을형간염간경화환자적골수간충질간세포(BMSCs)대자체외주혈림파세포증식급조절성T세포(Tregs)아군적영향.방법 종8례실대상성간경화환자골수중분리배양MSCs,채용류식세포의분석감정MSCs적순도,분리환자외주혈림파세포행최기형광소을선을산호박선아알지(CFDA SE)형광염색,재식물혈응소(PHA)자격하,배양기위자체혈청,자체외주혈림파세포여BMSCs공배양,류식세포술검측각조림파세포증식정황급CD4+ CD25+ CD127-Tregs빈수.결과 류식세포술현시:접촉배양조급비접촉배양조적세포증식교양성대조조명현감저(P균＜0.01),단역현저고우음성대조조(P균＜0.01),기CD4+ CD25+ CD127-Tregs빈솔현저고우양성급음성대조조(P균＜0.01);접촉배양조여비접촉배양조비교,림파세포증식급CD4+ CD25+ CD127-Tregs빈수차이균무통계학의의(P균＞0.05).결론 간경화환자BMSCs가억제자체외주혈림파세포증식、상조CD4+ CD25+ CD127-Tregs표체.
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.