Background：Mesenchymal stem cells （MSCs） transplantation has been proven to have therapeutic potential for acute liver failure （ALF）.However,the mechanism remains controversial.Recently,modulation of inflammation...Background：Mesenchymal stem cells （MSCs） transplantation has been proven to have therapeutic potential for acute liver failure （ALF）.However,the mechanism remains controversial.Recently,modulation of inflammation by MSCs has been regarded as a crucial mechanism.The aim of the present study was to explore the soluble cytokines secreted by MSCs and their therapeutic effects in ALF.Methods：MSCs isolated from Sprague-Dawley rats were identified by fluorescence-activated cell sorting analysis.Conditioned medium derived from MSCs （MSCs-CM） was collected and analyzed by a cytokine microarray.MSCs and MSCs-CM were transplanted into rats with D-galactosamine-induced ALF.Liver function,survival rate,histology,and inflammatory factors were determined.Exogenous recombinant rat interleukin （IL）-10,anti-rat IL-10 antibody,and AG490 （signal transducer and activator of transcription 3 [STAT3] signaling pathway inhibitor） were administered to explore the therapeutic mechanism of MSCs-CM.Statistical analysis was performed with SPSS version 19.0,and all data were analyzed by the independent-sample t-test.Results：There are statistical differences of the survival curve between ALF＋MSCs group and ALF＋Dulbecco＆#39;s modified Eagle＆#39;s medium （DMEM） group,as well as ALF＋MSCs-CM group and ALF＋DMEM group （all P 〈 0.05）.Serum alanine aminotransferase （ALT） level in the ALF＋MSCs and ALF＋MSCs-CM groups was lower than that in the ALF＋DMEM group （865.53＆#177;52.80 vs.1709.75＆#177;372.12 U/L and 964.72＆#177;414.59 vs.1709.75＆#177;372.12 U/L,respectively,all P 〈 0.05）;meanwhile,serum aspartate aminotransferase （AST） level in the ALF＋MSCs and ALF＋MSCs-CM groups was lower than that in the ALF＋DMEM group （2440.83＆#177;511.94 vs.4234.35＆#177;807.30 U/L and 2739.83＆#177;587.33 vs.4234.35＆#177;807.30 U/L,respectively,all P 〈 0.05）.Furthermore,MSCs or MSCs-CM treatment significantly reduced serum interferon-γ （IFN-γ）,IL-1β,IL-6 levels and increased 展开更多
[目的]探究微小RNA(microRNA,miR)-216a靶向调控酪氨酸蛋白激酶2/信号转导和转录激活因子3(Janus protein tyrosine protein kinase 2/signal transducer and activator of transcription 3,JAK2/STAT3)信号通路对鼻咽癌细胞增殖、侵袭...[目的]探究微小RNA(microRNA,miR)-216a靶向调控酪氨酸蛋白激酶2/信号转导和转录激活因子3(Janus protein tyrosine protein kinase 2/signal transducer and activator of transcription 3,JAK2/STAT3)信号通路对鼻咽癌细胞增殖、侵袭、自噬及血管生成的影响。[方法]Target Scan Human预测miR-216a与JAK2的结合位点,双荧光素酶实验验证miR-216a与JAK2是否结合;免疫组化染色检测正常鼻咽黏膜组织、鼻咽癌组织标本p-JAK2、p-STAT3的表达水平,并分析p-JAK2、p-STAT3表达水平与患者临床病理参数的关系。采用LipofectamineTM3000将miR-216a模拟物双链小RNA、阴性对照双链小RNA转染至CNE-2细胞构建过表达miR-216a细胞系,细胞分为对照组、miR-NC组、miR-216a组。将抑制剂双链小RNA,抑制剂对照双链小RNA转染至CNE-2细胞构建低表达miR-216a细胞系,细胞分为对照组、抑制miR-NC组、抑制miR-216a组。采用qRT-PCR检测细胞JAK2、STAT3 mRNA表达水平;MTT法、Transwell小室、皮下种植瘤模型、小管形成实验、吖啶橙染色分别检测细胞增殖、侵袭、移植瘤生长、血管形成和自噬泡形成的能力;Western blot检测JAK2、p-JAK2、STAT3、p-STAT3、微管相关蛋白轻链3-Ⅱ(microtubule-associated protein light chain 3-Ⅱ,CL3-Ⅱ)、酵母自噬相关基因6的哺乳动物同源体(autophagy related protein ATG6,Beclin1)、血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达水平。[结果]双荧光素酶实验验证了miR-216a对JAK2的靶向调控作用。鼻咽癌组织p-JAK2、p-STAT3阳性表达率(75.0%,68.3%)明显高于正常鼻咽黏膜组织(10%,13.3%)(P<0.05)。p-JAK2、p-STAT3表达越高,浸润深度越严重,越容易发生淋巴结和远处转移(P均<0.05)。细胞实验显示:与miR-NC组相比,miR-216a组细胞24、48、72、96 h吸光度值(0.32±0.03 vs 0.15±0.04、0.58±0.07 vs 0.24±0.06、0.86±0.10 vs 0.41±0.06、1.48±0.11 vs 0.75±0.07)、细胞侵袭数目(101.00±11.00 vs 22.67±5.77)、血�展开更多
基金This study was supported by grants from the National Natural Science Foundation of China，the Natural Science Foundation of Jiangsu Province,China
文摘Background：Mesenchymal stem cells （MSCs） transplantation has been proven to have therapeutic potential for acute liver failure （ALF）.However,the mechanism remains controversial.Recently,modulation of inflammation by MSCs has been regarded as a crucial mechanism.The aim of the present study was to explore the soluble cytokines secreted by MSCs and their therapeutic effects in ALF.Methods：MSCs isolated from Sprague-Dawley rats were identified by fluorescence-activated cell sorting analysis.Conditioned medium derived from MSCs （MSCs-CM） was collected and analyzed by a cytokine microarray.MSCs and MSCs-CM were transplanted into rats with D-galactosamine-induced ALF.Liver function,survival rate,histology,and inflammatory factors were determined.Exogenous recombinant rat interleukin （IL）-10,anti-rat IL-10 antibody,and AG490 （signal transducer and activator of transcription 3 [STAT3] signaling pathway inhibitor） were administered to explore the therapeutic mechanism of MSCs-CM.Statistical analysis was performed with SPSS version 19.0,and all data were analyzed by the independent-sample t-test.Results：There are statistical differences of the survival curve between ALF＋MSCs group and ALF＋Dulbecco＆#39;s modified Eagle＆#39;s medium （DMEM） group,as well as ALF＋MSCs-CM group and ALF＋DMEM group （all P 〈 0.05）.Serum alanine aminotransferase （ALT） level in the ALF＋MSCs and ALF＋MSCs-CM groups was lower than that in the ALF＋DMEM group （865.53＆#177;52.80 vs.1709.75＆#177;372.12 U/L and 964.72＆#177;414.59 vs.1709.75＆#177;372.12 U/L,respectively,all P 〈 0.05）;meanwhile,serum aspartate aminotransferase （AST） level in the ALF＋MSCs and ALF＋MSCs-CM groups was lower than that in the ALF＋DMEM group （2440.83＆#177;511.94 vs.4234.35＆#177;807.30 U/L and 2739.83＆#177;587.33 vs.4234.35＆#177;807.30 U/L,respectively,all P 〈 0.05）.Furthermore,MSCs or MSCs-CM treatment significantly reduced serum interferon-γ （IFN-γ）,IL-1β,IL-6 levels and increased
文摘[目的]探究微小RNA(microRNA,miR)-216a靶向调控酪氨酸蛋白激酶2/信号转导和转录激活因子3(Janus protein tyrosine protein kinase 2/signal transducer and activator of transcription 3,JAK2/STAT3)信号通路对鼻咽癌细胞增殖、侵袭、自噬及血管生成的影响。[方法]Target Scan Human预测miR-216a与JAK2的结合位点,双荧光素酶实验验证miR-216a与JAK2是否结合;免疫组化染色检测正常鼻咽黏膜组织、鼻咽癌组织标本p-JAK2、p-STAT3的表达水平,并分析p-JAK2、p-STAT3表达水平与患者临床病理参数的关系。采用LipofectamineTM3000将miR-216a模拟物双链小RNA、阴性对照双链小RNA转染至CNE-2细胞构建过表达miR-216a细胞系,细胞分为对照组、miR-NC组、miR-216a组。将抑制剂双链小RNA,抑制剂对照双链小RNA转染至CNE-2细胞构建低表达miR-216a细胞系,细胞分为对照组、抑制miR-NC组、抑制miR-216a组。采用qRT-PCR检测细胞JAK2、STAT3 mRNA表达水平;MTT法、Transwell小室、皮下种植瘤模型、小管形成实验、吖啶橙染色分别检测细胞增殖、侵袭、移植瘤生长、血管形成和自噬泡形成的能力;Western blot检测JAK2、p-JAK2、STAT3、p-STAT3、微管相关蛋白轻链3-Ⅱ(microtubule-associated protein light chain 3-Ⅱ,CL3-Ⅱ)、酵母自噬相关基因6的哺乳动物同源体(autophagy related protein ATG6,Beclin1)、血管内皮生长因子(vascular endothelial growth factor,VEGF)的表达水平。[结果]双荧光素酶实验验证了miR-216a对JAK2的靶向调控作用。鼻咽癌组织p-JAK2、p-STAT3阳性表达率(75.0%,68.3%)明显高于正常鼻咽黏膜组织(10%,13.3%)(P<0.05)。p-JAK2、p-STAT3表达越高,浸润深度越严重,越容易发生淋巴结和远处转移(P均<0.05)。细胞实验显示:与miR-NC组相比,miR-216a组细胞24、48、72、96 h吸光度值(0.32±0.03 vs 0.15±0.04、0.58±0.07 vs 0.24±0.06、0.86±0.10 vs 0.41±0.06、1.48±0.11 vs 0.75±0.07)、细胞侵袭数目(101.00±11.00 vs 22.67±5.77)、血�
