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-论文.pdf

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1、262·中华实验和临床病毒学杂志2014年8月第28卷第4期ChineseJExpClinVirol，August2014，Vo1．28，No．4．论著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郭春辉，韩兰秀，万美容，甘建和，邓国炯【摘要】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(CFDASE)荧光染色，在植物血凝素(PHA)刺激下，

2、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CD4CD25CD127。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．叭)，但亦显著高于阴性对照组(P均<0．O1)，其CD4CD25CD127。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CD4CD25CD127。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CD4CD25CD127一Tregs表达。【主题

3、词】肝炎，乙型，肝硬化；骨髓细胞；T细胞ImmuneregulatoryefectofbonemarrowmesenchymalstemcellsofpatientwithhepatitisBvirusrelateddecompensatedcirrhosis．GuoChunhui，HanLanxiu，WanMeirong，GanJianhe，DengGuojiong．DepartmentofinfectiousDiseases．JiangyinHospitalAffiliatedtoSoutheastUniversity，JiangsuJiangyin214400，Chin

4、aCorrespondingauthor：Dengguojiong，Email：DengguojiongO07@163．com【Abstract】0bjectiveTostudytheeffectofbonemarrowmesenchymalstemcells(BMSCs)onautologousperipheralbloodlymphocyteproliferationandregulatoryTcells(Tregs)subsetsinvitroinpatientswithhepatitisBvirusrelateddecompensatedcirrhosis．Metho

5、dsMSCswereisolatedandexpandedfromhumanbonemarrowbloodofeightpatientswithdecompensatedcirrhosis．ThepurityofMSCswasidentifiedbyflowcytometry．1ymphocyteswereisolatedfromtheperipheralbloodofpatientsandstainedwithcarboxyfluoresceindiacetatesuccinimidylester(CFDASE)．TheBMSCsandperipheralbloodlymp

6、hocytesfrompatientswereaddedintowellscontainingautologousseruminthepresenceofphytohemagglutinin(PHA)．1ymphocytesproliferationrateandCD4CD25CD127一Tregsfrequencyweredetectedbyflowcytometry．ResultsFlowcytometricanalysisshowedthatlymphocyteproliferationincontactCO-cuhuregroupandnon—contactCO—cu

7、lturegroupweremuchlowerthanpositivecontrol(allP<0．01)，andsignificantlyhigherthannegativecontrol(allP<0．01)，whileCD4CD25CD127。TregsincontactCO—culturegroupandnon-contactCO-culturegroupweremuchhigherthanpositivecontrolandnegativecontrol(allP<0．O1)．Besides，