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Reg Ⅲγ修饰HuMSCs对柯萨奇B3病毒诱导的心肌炎炎性细胞浸润及心脏功能的影响
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作者 蹇强 李丹 +1 位作者 程玮 孙敏 《中西医结合心脑血管病杂志》 2023年第8期1415-1422,共8页
目的:观察胰岛再生源蛋白Ⅲγ(RegⅢγ)修饰人脐带间充质干细胞(HuMSCs)对柯萨奇B3病毒(CVB3)诱导的小鼠心肌炎炎性细胞浸润及心脏功能的影响。方法:从新鲜脐带组织中提取HuMSCs,倒置显微镜观察形态,流式细胞仪检测细胞表面抗原表达;采... 目的:观察胰岛再生源蛋白Ⅲγ(RegⅢγ)修饰人脐带间充质干细胞(HuMSCs)对柯萨奇B3病毒(CVB3)诱导的小鼠心肌炎炎性细胞浸润及心脏功能的影响。方法:从新鲜脐带组织中提取HuMSCs,倒置显微镜观察形态,流式细胞仪检测细胞表面抗原表达;采用含过表达RegⅢγ的慢病毒感染HuMSCs,荧光显微镜观察绿色荧光蛋白表达,实时荧光定量聚合酶链式反应(qRT-PCR)检测RegⅢγ mRNA表达水平。将40只小鼠按照随机数字表法分为对照组、模型组、HuMSCs组和RegⅢγ-HuMSCs组,每组10只。模型组、HuMSCs组和RegⅢγ-HuMSCs组采用腹腔注射CVB3建立心肌炎模型,HuMSCs组和RegⅢγ-HuMSCs组再分别尾静脉注射HuMSCs、RegⅢγ-HuMSCs。14 d后,采用高分辨率小动物超声成像系统记录小鼠心脏超声指标;酶联免疫吸附法(ELISA)检测各组血清白细胞介素(IL)-2、IL-4、IL-6、IL-10和IL-23含量;苏木精-伊红(HE)染色观察心肌组织病理损伤情况,末端脱氧核苷酸转移酶介导的dUTP缺口末端标记测定法(TUNEL)检测心肌组织细胞凋亡水平,免疫组织化学染色测定心肌组织T淋巴细胞表面抗原(CD4、CD8)和巨噬细胞特异性抗体(CD68)的表达。结果:分离的细胞呈典型纺锤形,细胞表面抗原CD44、CD73、CD90呈高表达,CD34、CD45、人类白细胞抗原-DR蛋白(HLA-DR)呈低表达,说明成功分离到HuMSCs。经过慢病毒感染的HuMSCs有明显绿色荧光蛋白表达,RegⅢγ mRNA表达水平高于未感染的细胞(P<0.05),说明成功得到RegⅢγ修饰的HuMSCs。与对照组比较,模型组小鼠射血分数(EF)、缩短分数(FS)降低,左室舒张末期容积(LVEDV)和左室收缩末期容积(LVESV)升高,血清IL-2、IL-6和IL-23含量增加,IL-4和IL-10含量减少,心肌组织内炎性细胞浸润明显,TUNEL阳性率增加,CD4、CD8及CD68蛋白表达均呈强阳性(P<0.05);与模型组比较,HuMSCs组和RegⅢγ-HuMSCs组小鼠EF和FS升高,LVEDV和LVESV降低,血清IL-2、IL-6和IL-23含量减少,IL-4和IL-10含量增加,心肌组织炎性细胞浸润减少,TUNEL阳性率减少,CD4、CD8及CD68蛋白表达均下降(P<0.05);相较于HuMSCs组,RegⅢγ-HuMSCs组小鼠EF和FS升高,LVEDV和LVESV降低,血清IL-2、IL-6和IL-23含量减少,IL-4和IL-10含量增加,心肌组织未见明显炎性细胞浸润,TUNEL阳性率减少,CD4、CD8及CD68蛋白表达下降(P<0.05)。结论:RegⅢγ修饰的HuMSCs可减轻CVB3诱导的小鼠心肌炎心肌损伤与炎性细胞浸润,改善心脏功能。 展开更多
关键词 心肌炎 柯萨奇B3病毒 胰岛再生源蛋白Ⅲγ 人脐带间充质干细胞 humscs 炎性细胞 小鼠 实验研究
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胰岛素样生长因子-I对人脐带间充质干细胞增殖影响的量效关系 被引量:1
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作者 徐斌 唐军 刘毅 《中国美容医学》 CAS 2011年第8期1247-1250,共4页
目的:探讨不同剂量胰岛素样生长因子-I对人脐带间充质干细胞(hUCMSCs)增殖的影响及其量效关系。方法:取第2代hUCMSCs接种于96孔板,分为7组(G1,G2…,G7)。前5组分别加入含不同剂量IGF-I(40,60,80,100,120μg/L)的体积分数为0.02的胎牛血... 目的:探讨不同剂量胰岛素样生长因子-I对人脐带间充质干细胞(hUCMSCs)增殖的影响及其量效关系。方法:取第2代hUCMSCs接种于96孔板,分为7组(G1,G2…,G7)。前5组分别加入含不同剂量IGF-I(40,60,80,100,120μg/L)的体积分数为0.02的胎牛血清培养液;第6组加入体积分数为0.05的胎牛血清培养液做为阳性对照,第7组加入体积分数为0.02的胎牛血清培养液做为阴性对照,采用MTT法观察不同剂量的IGF-I对细胞增殖的影响。结果:①第3天、第6天,G1组、G2组和G7组之间差异无显著性(P>0.05);G3组、G4组和G5组之间差异无显著性(P>0.05);②第3天,G6组增殖高于G1组与G2组,低于G3组、G4组和G5组,差异有显著性(P<0.000或<0.006);第4天,G6组增殖高于其余各组,差异有显著性(P<0.000或<0.006)。③第3天、第6天,G1组、G2组和G6组低于G3组,G4组,G5组,差异有显著性(P<0.000);G6组增殖高于G7组,统计分析差异有显著性(P<0.000)。结论:外源性IGF-I对hUCMSCs增殖的促进作用,随IGF-I浓度增高而有增强趋势,100μg/L为其较适合浓度,为实现hUCMSCs体外快速扩增提供适宜的条件。 展开更多
关键词 humscs IGF-I 组织工程
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Human umbilical cord mesenchymal stem cells as cell carriers in the treatment of gliomas: research status and prospects
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作者 Qianxu Jin Zongmao Zhao 《Journal of Translational Neuroscience》 2020年第2期22-33,共12页
Human umbilical cord mesenchymal stem cells(HuMSCs)have the multi-difFerentiation potential to differentiate into various types of cells without immune rejection.They are considered to be an ideal source of neural ste... Human umbilical cord mesenchymal stem cells(HuMSCs)have the multi-difFerentiation potential to differentiate into various types of cells without immune rejection.They are considered to be an ideal source of neural stem cells and also an ideal cell carrier for gene therapy.Because of the invasive growth of brain gliomas,most of them have no obvious boundaries with normal brain tissues.It is difficult to completely remove them by surgery and the remaining cells become the main source of tumor recurrence.In recent years,gene therapy has become a new method for the treatment of gliomas.The vector carrying the target gene is introduced into HuMSCs in a certain way to correct gene defects or play other roles.The differentiation potential of HuMSCs makes it an ideal source of nerve cells to play a greater role in gene therapy of glioma.Therefore,this article reviews the current status and prospects of HuMSCs as cell carriers in the treatment of glioma. 展开更多
关键词 human umbilical cord mesenchymal stem cells(humscs) GLIOMA cell carrier gene therapy targeted therapy
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Identification of microRNAs and messenger RNAs involved in human umbilical cord mesenchymal stem cell treatment of ischemic cerebral infarction using integrated bioinformatics analysis 被引量:13
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作者 Yin-Meng Qu Xin Sun +3 位作者 Xiu-Li Yan Hang Jin Zhen-Ni Guo Yi Yang 《Neural Regeneration Research》 SCIE CAS CSCD 2019年第9期1610-1616,共7页
In recent years,a large number of differentially expressed genes have been identified in human umbilical cord mesenchymal stem cell(hUMSC)transplants for the treatment of ischemic cerebral infarction.These genes are i... In recent years,a large number of differentially expressed genes have been identified in human umbilical cord mesenchymal stem cell(hUMSC)transplants for the treatment of ischemic cerebral infarction.These genes are involved in various biochemical processes,but the role of microRNAs(miRNAs)in this process is still unclear.From the Gene Expression Omnibus(GEO)database,we downloaded two microarray datasets for GSE78731(messenger RNA(mRNA)profile)and GSE97532(miRNA profile).The differentially expressed genes screened were compared between the hUMSC group and the middle cerebral artery occlusion group.Gene ontology enrichment and pathway enrichment analyses were subsequently conducted using the online Database for Annotation,Visualization,and Integrated Discovery.Identified genes were applied to perform weighted gene co-suppression analyses,to establish a weighted co-expression network model.Furthermore,the protein-protein interaction network for differentially expressed genes from turquoise modules was built using Cytoscape(version 3.40)and the most highly correlated subnetwork was extracted from the protein-protein interaction network using the MCODE plugin.The predicted target genes for differentially expressed miRNAs were also identified using the online database starBase v3.0.A total of 3698 differentially expressed genes were identified.Gene ontology analysis demonstrated that differentially expressed genes that are related to hUMSC treatment of ischemic cerebral infarction are involved in endocytosis and inflammatory responses.We identified 12 differentially expressed miRNAs in middle cerebral artery occlusion rats after hUMSC treatment,and these differentially expressed miRNAs were mainly involved in signaling in inflammatory pathways,such as in the regulation of neutrophil migration.In conclusion,we have identified a number of differentially expressed genes and differentially expressed mRNAs,miRNA-mRNAs,and signaling pathways involved in the hUMSC treatment of ischemic cerebral infarction.Bioinformatics and interaction analyses can provide novel clues for further research into hUMSC treatment of ischemic cerebral infarction. 展开更多
关键词 nerve REGENERATION ischemic cerebral infarction human umbilical cord mesenchymal STEM CELL TREATMENT bioinformatics analysis DIFFERENTIALLY EXPRESSED genes DIFFERENTIALLY EXPRESSED mRNAs inflammatory response STEM CELL therapy weighted gene co-suppression analysis WGCNA protein-protein interaction network PPI hUMSC neural REGENERATION
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Role of miR-128/216a Regulating Isl1 Expression during Differentiation of Human Umbilical Cord Mesenchymal Stem Cells into Insulin-Producing Cells
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作者 MU Tianchi WANG Tao +2 位作者 GAO Zhenyu PAN Xin LIU Yingxue 《Wuhan University Journal of Natural Sciences》 CAS CSCD 2023年第2期177-184,共8页
Islet-1(Isl1),a LIM homeodomain protein,is expressed in the embryonic pancreatic epithelium.As a key transcription factor,Isl1 can not only regulate insulin gene expression in normal glucose condition but also maintai... Islet-1(Isl1),a LIM homeodomain protein,is expressed in the embryonic pancreatic epithelium.As a key transcription factor,Isl1 can not only regulate insulin gene expression in normal glucose condition but also maintainβ-cell function and impact pancreaticβ-cell target genes.Some experiments have suggested that Micro RNA(miRNA)can play a critical role during the induction of insulinproducing cells(IPCs).However,it is unclear whether miRNA may regulate Isl1 expression during differentiation of human umbilical cord mesenchymal stem cells(HUMSCs)into IPCs.In this investigation,we induced HUMSCs into IPCs with a modified two-step protocol,activin A,retinoic acid(step1)and conophylline,nicotinamide(step2).To find the miRNA regulating Isl1 expression,we respectively used Target Scan,miRDB and RNAhybrid to predict and got the result,miR-128 and miR-216a.The miRNAs can inhibit Isl1 expression by dual luciferase assay.The results of real-time Polymerase Chain Reaction(PCR)showed that Isl1 expression level was almost reciprocal to that of miR-128 and miR-216a during differentiation of HUMSCs into IPCs.Furthermore,over-expression of miR-128 or miR-216a downregulated expression levels of Isl1 and Maf A.Therefore,miR-128 or miR-216a may regulate expression of islet-specific transcription factors to control differentiation of HUMSCs into IPCs. 展开更多
关键词 microRNA(miRNA) Isl1 human umbilical cord mesenchymal stem cells(humscs) insulin-producing cells(IPCs)
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