心肺复苏干细胞移植后肿瘤坏死因子α诱导蛋白6的表达
Expression of tumor necrosis factor-α-induced protein 6 after transplantation of mesenchymal stem cells in a rat model of cardiopulmonary resuscitation
摘要目的 探讨骨髓间充质于细胞(MSCs)移植对心肺复苏(CPR)大鼠脑肿瘤坏死因子×诱导蛋白6 (TSG-6)表达的影响.方法 SD大鼠随机(随机数字法)分为假手术组(Sham组)、磷酸盐缓冲液注射组(PBS对照组)和MSCs移植组.通过窒息法诱导大鼠心搏骤停后进行CPR,复苏后2h,PBS对照组和MSCs移植组分别静脉注射PBS与MSCs.CPR后1d、3d和7d,对各组大鼠进行神经功能缺损评分(NDS),酶联免疫吸附法(ELISA)检测血清S-100B,免疫荧光检测MSCs在脑内分布和TSG-6的表达,实时RT-PCR检测脑组织TSG-6和促炎症因子的表达水平,Western blot检测脑组织中性粒细胞弹性蛋白酶(NE)表达水平.组间比较采用方差分析.结果 CPR后3d和7d,MSCs移植组NDS均高于PBS对照组(P<0.01),血清S-100B均低于PBS对照组(P<0.01).DAPI标记的MSCs迁移到损伤脑组织,一些DAPI阳性细胞表达TSG-6.CPR后3d和7d,MSCs移植组脑组织TSG-6表达均高于PBS对照组(P<0.01),而NE和促炎症因子的表达均低于PBS对照组(P<0.05).结论 静脉MSCs移植可能通过TSG-6抑制CPR后大鼠脑组织炎症反应并改善神经功能.
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abstractsObjective To investigate the effects of bone marrow mesenchymal stem cells (MSCs)treatment on TSG-6 in a rat model of cardiopulmonary resuscitation (CPR).Methods Sprague Dawley (SD) rats were randomly (random number) divided into sham group,phosphate buffer solution (PBS)-treated group and MSCs-treated group.Animals were subjected to asphyxial cardiac arrest followed by CPR.In PBS-treated group or MSCs-treated group,animals were injected intravenously with PBS or MSCs at 2h after resuscitation.Neurological deficit scores (NDS) were assessed at 1,3 and 7 d after CPR.Serum S-100B was assayed using enzyme linked immunosorbent assay (ELISA).Immunofluorescence was performed to detect donor MSCs and the expression of TSG-6 in brain.TSG-6 and proinflammatory cytokines in brain were assayed using real time reverse transcription-polymerase chain reaction (RT-PCR).Western blot analysis was performed to measure the levels of neutrophil elastase (NE) in brain.Multiple comparisons were made by analysis of variance.Results At 3d and 7d,MSCs-treated group demonstrated higher NDS than PBS-treated group (P < 0.01),and serum S-100B levels significantly reduced in MSCs-treated group compared with PBS-treated group (P < 0.01).DAPI-labeled MSCs migrated into the ischemic brain and some DAPI + cells colocalized with TSG-6.Compared with PBS-treated group,MSCs treatment significantly up-regulated the expression of TSG-6 and reduced the expression of NE and proinflammatory cytokines in brain at 3 d and 7 d after CPR (P < 0.05).Conclusion Systemically administered MSCs suppressed inflammatory responses in brain after CPR and improved neurological function in rats possibly via induction of TSG-6.
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