APPswe小鼠β淀粉样斑块与胶质细胞激活相关性研究

摘要 目的：通过检测阿尔茨海默病（Alzheimer''s Disease,AD）APPswe转基因小鼠前额叶皮质（prefrontal cortex, PFC）中β-淀粉样蛋白（β-amyloid, Aβ）与胶质纤维酸性蛋白(glial fibrillary acidic protein, GFAP)和离子钙结合衔接分子1（Ionized calcium binding adaptor molecule-1,Iba-1）的表达相关性,进一步明确AD中β淀粉样斑块在神经胶质细胞激活中的重要作用。方法：选取9只12月龄APPswe AD小鼠,使用免疫组织化学法检测PFC中Aβ、GFAP以及Iba-1表达,并分析Aβ与GFAP和Iba-1水平的相关性;使用免疫荧光双标法分别评价不同大小Aβ斑块分别与GFAP和Iba-1的共染情况。结果：免疫组织化学结果显示,AD小鼠前额叶皮质中Aβ水平高,则GFAP和Iba-1信号水平也较高;反之,Aβ水平低,GFAP和Iba-1的表达水平也较低。Pearson相关性分析结果表明Aβ水平与GFAP（R=0.6677,P<0.05）和Iba-1（R=0.8257,P<0.05）的水平呈正相关,且与GFAP相比,Iba-1显示出与Aβ水平更高的表达相关性。与免疫组织化学结果一致,免疫荧光双标法结果亦表明小鼠PFC中较大的Aβ斑块所在区域GFAP或Iba-1荧光信号强度及范围亦大于较小的Aβ斑块所在区域GFAP或Iba-1荧光信号强度。结论：AD小鼠前额叶皮质中Aβ水平与GFAP和Iba-1的表达量呈正相关,表明Aβ形成在神经胶质细胞的激活中发挥重要作用。     

成年大鼠吻侧迁移流的神经发生

目的探讨成年SD大鼠吻侧迁移流(rostral migratory stream,RMS)的神经发生。方法成年6周龄SD大鼠被处死。矢状位切片,免疫组织化学染色观察RMS区微管相关蛋白双皮质素(doublecortin,DCX)及胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)的表达及DCX/GFAP、DCX/p-CREB的共表达情况。结果在RMS区的垂直臂、肘部、水平臂均有DCX阳性细胞和GFAP阳性细胞,RMS区有GFAP/DCX和p-CREB/DCX共表达细胞。结论吻侧迁移流有广泛的神经元前体细胞及星形胶质细胞标记物表达;迁移神经元标记物可表达于星形胶质细胞,且神经元的迁移受到CREB信号通路的调控。     

白藜芦醇对脊髓损伤小鼠神经元凋亡及Fas/FasL等凋亡相关蛋白表达的影响

摘要 目的：研究白藜芦醇对脊髓损伤（Spinal cord injury, SCI）小鼠脊髓组织神经元凋亡和凋亡相关蛋白表达的影响。方法：21只雌性C57BL/6小鼠,6-8周龄,随机分为三组：Sham组（假手术对照组）,SCI组（脊髓损伤模型）和Resveratrol组（白藜芦醇治疗的脊髓损伤模型）,每组7只。通过Basso小鼠量表（BMS）评估小鼠后肢运动功能、HE染色评估小鼠脊髓病变面积、尼氏染色检测脊髓组织神经元数目、TUNEL染色检测凋亡细胞数目。通过酶联免疫试剂盒检测脊髓组织髓过氧化物酶（myeloperoxidase,MPO）、肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）、白细胞介素（interleukin, IL）-6和IL-8蛋白表达水平。通过免疫印迹法检测脊髓组织凋亡相关蛋白Fas、FasL、caspase 3和caspase 8表达水平。结果：与Sham组小鼠相比,SCI模型小鼠脊髓组织病变面积和MPO活性均显著增加（P<0.05）,但经白芦藜醇治疗后的Resveratrol组SCI小鼠脊髓组织病变面积和MPO活性较SCI组小鼠显著降低（P<0.05）。与Sham组小鼠相比,SCI模型小鼠BMS评分显著降低（P<0.05）,但经白芦藜醇治疗后的Resveratrol组SCI小鼠BMS评分较SCI组小鼠显著升高（P<0.05）。与Sham组小鼠相比,SCI模型小鼠脊髓组织神经元丢失和凋亡均显著增加（P<0.05）,但经白芦藜醇治疗后的Resveratrol组SCI小鼠脊髓组织病神经元丢失和凋亡较SCI组小鼠显著降低（P<0.05）。与Sham组小鼠相比,SCI模型小鼠脊髓组织Fas、FasL、caspase 3、caspase 8、TNF-α、IL-6和IL-8蛋白表达水平均显著升高（P<0.05）,但经白芦藜醇治疗后均显著降低（P<0.05）。结论：白芦藜醇可显著降低脊髓损伤小鼠脊髓组织神经元凋亡,其机制可能与抑制脊髓损伤小鼠脊髓组织炎症和炎症引起的凋亡蛋白表达有关。     

人参皂苷Rh2对大鼠C6胶质瘤细胞Siah-1、Synaptophysin、MMP9及VEGF表达的影响

摘要 目的：探讨人参皂苷Rh2对大鼠C6胶质瘤细胞Siah-1、突触素（Synaptophysin）、基质金属蛋白酶-9（MMP-9）血管内皮生长因子（VEGF）表达的影响。方法：将大鼠C6胶质瘤细胞分为对照组、人参皂苷Rh2低剂量组（16 μg/mL）、人参皂苷Rh2中剂量组（32 μg/mL）、人参皂苷Rh2高剂量组（48 μg/mL），CCK-8法和平板克隆实验检测细胞增殖；流式细胞术检测细胞凋亡；Transwell检测细胞侵袭；实时荧光定量-聚合酶链式反应（qRT-PCR）检测C6胶质瘤细胞中VEGF、Siah-1、Synaptophysin、MMP-9 mRNA表达；蛋白质印迹法（Western blot）检测C6胶质瘤细胞中VEGF、Siah-1、Synaptophysin、MMP-9蛋白表达。结果：与对照组比较，人参皂苷Rh2低剂量组、人参皂苷Rh2中剂量组、人参皂苷Rh2高剂量组大鼠C6胶质瘤细胞OD₄₅₀值（24 h、48 h）、克隆形成率、细胞侵袭数、VEGF、Synaptophysin、MMP-9 mRNA及蛋白表达降低，细胞凋亡率、Siah-1 mRNA及蛋白表达升高，且呈剂量依赖性（P<0.05）。结论：人参皂苷Rh2可能通过上调Siah-1，下调VEGF、Synaptophysin、MMP-9表达来抑制大鼠C6胶质瘤细胞增殖与侵袭，促进细胞凋亡。     

SPARCL1通过MEK/ERK信号通路调节非小细胞肺癌细胞的增殖、凋亡和侵袭

摘要 目的：分析富含半胱氨酸的酸性分泌蛋白类似蛋白1（SPARCL1）对非小细胞肺癌（NSCLC）细胞增殖、凋亡、侵袭的影响,并探讨分裂原活化抑制剂（MEK）/细胞外调节蛋白激酶（ERK）通路在其中发挥的作用。方法：收集2019年9月~2021年6月期间本院接受手术治疗的84例NSCLC患者癌组织与相应癌旁组织,实时定量逆转录聚合酶链反应（qRT-PCR）法测定并比较各组织以及正常肺上皮细胞HBEpiC、NSCLC细胞A549、HCC827、H1299、H292中SPARCL1 信使RNA（mRNA）表达水平,选取A549、HCC827培养并分组,分为对照组、NC siRNA组、SPARCL1 siRNA组、U0126组（MEK/ERK特异性抑制剂）、SPARCL1 siRNA加U0126组,细胞计数法（CCK8）以及平板克隆法测定A549、HCC827细胞增殖,流式细胞仪测定A549、HCC827细胞凋亡,Transwell小室法测定A549、HCC827细胞侵袭能力,蛋白质印迹法（western blot）检测SPARCL1、p-MEK、MEK、p-ERK1/2、ERK1/2蛋白表达。结果：SPARCL1在NSCLC组织中mRNA表达水平低于癌旁组织（P＜0.05）;与HBEpiC细胞相比,NSCLC细胞A549、HCC827、H1299、H292细胞中SPARCL1 mRNA表达水平降低（P＜0.05）;与对照组相比,SPARCL1 siRNA组A549、HCC827细胞SPARCL1 mRNA表达水平与蛋白表达、凋亡率降低（P＜0.05）,OD₄₅₀、克隆形成数、侵袭细胞数、p-MEK/MEK、p-ERK1/2/ERK1/2蛋白表达升高（P＜0.05）,U0126组A549、HCC827细胞SPARCL1 mRNA表达水平与蛋白表达、凋亡率升高（P＜0.05）,OD₄₅₀、克隆形成数、侵袭细胞数、p-MEK/MEK、p-ERK1/2/ERK1/2蛋白表达降低（P＜0.05）;与SPARCL1 siRNA组相比,SPARCL1 siRNA加U0126组A549、HCC827细胞SPARCL1 mRNA表达水平与蛋白表达、凋亡率升高（P＜0.05）,OD₄₅₀、克隆形成数、侵袭细胞数、p-MEK/MEK、p-ERK1/2/ERK1/2蛋白表达降低（P＜0.05）。结论：SPARCL1可能通过调控MEK/ERK通路影响NSCLC A549、HCC827细胞增殖、侵袭与凋亡。     

基于自噬与凋亡机制研究左归丸对化疗损伤颗粒细胞及膜细胞的影响

目的 探讨左归丸含药血清对化疗损伤性颗粒细胞和膜细胞的影响及作用机制。方法 制备左归丸含药血清,培养大鼠卵巢颗粒细胞和膜细胞,使用磷酰胺氮芥造模分组后给药。CCK-8法测定颗粒细胞和膜细胞存活率,实时荧光定量PCR法（RT-PCR）及蛋白质免疫印迹法（Western blot）分别检测卵巢自噬启动因子Beclin-1、微管结合蛋白轻链3（LC3B）、自噬受体蛋白p62、凋亡蛋白Bax、Caspase3在转录水平和翻译水平上的表达。结果 10%左归丸含药血清对于细胞存活的挽救率最高。Beclin-1、LC3B、Bax、Caspase3在磷酰胺氮芥作用的颗粒细胞和膜细胞中,相对于空白对照组有高表达（P<0.05）,10%左归丸含药血清可下调上述蛋白质在模型组中的表达（P<0.05）;然而受体蛋白p62较空白对照组升高（P<0.05）,10%左归丸含药血清可上调模型组p62的表达（P<0.05）。此外,在颗粒细胞实验组中,激活或抑制自噬途径后,自噬相关蛋白的表达在发生相应改变的同时,凋亡相关蛋白的表达也会发生相应改变。结论 磷酰胺氮芥可通过促进凋亡、激活自噬/溶酶体降解途径的机制损伤颗粒细胞和膜细胞。10%左归丸含药血清能缓解由此带来的损伤,同时影响了颗粒细胞和膜细胞自噬和凋亡过程。在磷酰胺氮芥损伤颗粒细胞的过程和10%左归丸含药血清缓解其损伤过程中均存在自噬与凋亡串流（cross-talk）。     

大黄酸调节Ras/ERK信号通路对肝细胞癌细胞增殖、迁移和侵袭的影响

摘要 目的：探讨大黄酸调节大鼠肉瘤蛋白（Ras）/胞外信号调控激酶（ERK）信号通路对肝细胞癌（HCC）细胞增殖、迁移和侵袭的影响。方法：使用不同浓度（0、12.50、25、50、100、150、200 mol/L）大黄酸处理HepG2细胞,检测细胞活性,筛选最佳大黄酸浓度。将细胞分为对照组、大黄酸低、中、高浓度组、大黄酸高浓度+Ras/ERK激活剂组（大黄酸高浓度+ML-099组）,分别检测各组细胞集落形成数、划痕愈合率、细胞侵袭数和Ras、p-ERK、ERK蛋白表达。结果：大黄酸以浓度和时间依赖性降低HepG2细胞活性（P<0.05）,选用25、50、100 mol/L处理HepG2细胞24 h用于后续实验;与对照组比较,大黄酸低、中、高浓度组细胞集落形成数、G0/G1细胞比例、细胞划痕愈合率、细胞侵袭数和原癌基因（c-Myc）、细胞周期蛋白D1（CyclinD1）、Ras、p-ERK/ERK蛋白表达呈浓度依赖性降低,S期和G2/M细胞比例、p53蛋白表达呈浓度依赖性增加（P<0.05）;与大黄酸高浓度组比较,大黄酸高浓度+ML-099组细胞集落形成数、G0/G1细胞比例、细胞划痕愈合率、细胞侵袭数和c-Myc、CyclinD1、Ras、p-ERK/ERK蛋白表达显著增加,S期和G2/M细胞比例、p53蛋白表达显著降低（P<0.05）。结论：大黄酸可能通过抑制Ras/ERK信号通路抑制HCC细胞增殖、迁移和侵袭。     

rt-PA动脉溶栓联合血管内支架成形术对早期急性脑梗死患者纤溶系统和血清神经功能损伤指标的影响

摘要 目的：探讨重组人组织型纤溶酶原激活物（rt-PA）动脉溶栓联合血管内支架成形术治疗早期急性脑梗死（ACI）的疗效及对纤溶系统和血清神经功能损伤指标的影响。方法：选取我院2018年9月~2021年3月间接收的80例早期ACI患者。根据随机数字表法分为对照组38例（rt-PA动脉溶栓治疗）和研究组42例（rt-PA动脉溶栓联合血管内支架成形术治疗）。对比两组血管再通情况、纤溶系统和血清神经功能损伤指标变化、日常生活能力量表（ADL）及美国国立卫生研究院卒中量表（NIHSS）评分,观察两组预后情况。结果：研究组的血管完全再通率高于对照组（P<0.05）。研究组治疗后1 d、治疗后7 d、治疗后3个月NIHSS评分低于对照组,ADL评分高于对照组（P<0.05）。研究组治疗7 d后血管性假血友病因子（vWF）、血浆纤溶酶原激活物抑制剂-1（PAI-1）低于对照组,组织型纤溶酶原激活剂（tPA）高于对照组（P<0.05）。研究组治疗7 d后胶质纤维酸性蛋白（GFAP）、神经元特异性烯醇化酶（NSE）、S100β蛋白低于对照组（P<0.05）。研究组患者的再发脑梗死率、死亡率低于对照组（P<0.05）。结论：rt-PA动脉溶栓联合血管内支架成形术治疗早期ACI患者,可改善患者近远期疗效和预后,有效调节纤溶系统,减轻机体神经功能损伤,提高患者日常生活能力。     

电针对脊髓损伤星形胶质细胞增生及其NGF表达的影响

目的研究脊髓损伤后电针治疗对星形胶质细胞增生及其内源性神经生长因子(nerve growth factor,NGF)表达的影响.方法选用成年雌性Wistar大鼠,随机分为3组.A组为正常对照组,B组、C组为下胸段脊髓不完全损伤.B组损伤后不治疗,C组损伤后给予督脉电针治疗.损伤后3 d、1 、2或4周应用免疫组化染色分别观察损伤脊髓胶质原纤维酸性蛋白(glial fibroblast acid protein,GFAP)和NGF表达的变化.结果 B组术后3 d,GFAP阳性细胞明显增多, 2周后开始减少,4周时仍有较多的阳性细胞;C组GFAP阳性细胞明显少于B组,1周时达高峰.脊髓损伤后NGF表达呈逐渐增加的趋势.C组NGF的表达明显高于B组,且一直保持在较高水平.NGF阳性细胞大部分与GFAP阳性细胞形态相似.结论电针治疗能减少星形胶质细胞增生,促进内源性NGF的合成,从而创造了有利于神经再生的微环境.     

MMPs抑制剂对结肠癌细胞凋亡、免疫功能及炎性因子的影响

摘要 目的：探究基质金属蛋白酶（Matrix metalloproteinases,MMPs）抑制剂对结肠癌细胞凋亡、免疫功能及炎性因子的影响。方法：取SW480结肠癌细胞,随机分为低表达组（将MMPs抑制剂慢病毒质粒与SW480细胞混合培养）,空白对照组（SW480细胞与慢病毒包装质粒混合培养）。采用流式细胞法、免疫细胞化学法、酶联免疫吸附（Enzyme-linked immunosorbent assay,ELISA）法、实时荧光定量PCR（real-time fluorescence quantitative PCR, qRT-PCR）法、Hoechst 33258 荧光染色法检测SW480细胞凋亡,TNF-α、IL-6蛋白阳性表达、免疫球蛋白表达水平、MMP-9 mRNA表达量。结果：空白对照组与低表达组相比较,低表达组SW480细胞内MMP-9 mRNA表达量显著下降(P<0.05),说明转染成功。与空白对照组相比较,低表达组SW480细胞凋亡数量显著上升(P<0.05)。低表达组细胞的细胞核出现核固缩的概率显著高于空白对照组（P<0.05）。与空白对照组相比较,低表达组SW480细胞IL-2表达水平显著升高（P＜0.05）,IL-4、TGF-β1、TIM-1表达水平显著降低（P＜0.05）。SW480细胞内的MMP-9 mRNA与IL-2呈现负相关性（r=-0.723,P=0.007）,MMP-9 mRNA与IL-4、TGF-β1及TIM-1均呈现负相关性（均P＜0.05）。空白对照组SW480细胞中TNF-α、IL-6蛋白阳性数最高,低表达组SW480细胞中TNF-α、IL-6蛋白阳性数最低,与空白对照组相比较,低表达组SW480细胞中TNF-α、IL-6阳性数显著下降(均P<0.05)。结论：MMPs抑制剂可促进SW480细胞凋亡,改善免疫功能,降低炎性介质的表达水平。     

miR-31 promotes neural stem cell proliferation and restores motor function after spinal cord injury

This study aims to examine whether miR-31 promotes endogenous NSC proliferation and be used for spinal cord injury management. In the present study, the morpholino knockdown of miR-31 induced abnormal neuronal apoptosis in zebrafish, resulting in impaired development of the tail. miR-31 agomir transfection in NSCs increased Nestin expression and decreased ChAT and GFAP expression levels. miR-31 induced the proliferation of mouse NSCs by upregulating the Notch signaling pathway, and more NSCs entered G1; Notch was inhibited by miR-31 inactivation. Injection of a miR-31 agomir into mouse models of spinal cord injury could effectively restore motor functions after spinal cord injury, which was achieved by promoting the proliferation of endogenous NSCs. After the injection of a miR-31 agomir in spinal cord injury mice, the expression of Nestin and GFAP increased, while GFAP expression decreased. In conclusion, the zebrafish experiments prove that a lack of miR-31 will block nervous system development. In spinal cord injury mouse models, miR-31 overexpression might promote spinal cord injury repair.     

Promotion of Survival and Differentiation of Neural Stem Cells with Fibrin and Growth Factor Cocktails after Severe Spinal Cord Injury

Neural stem cells (NSCs) can self-renew and differentiate into neurons and glia. Transplanted NSCs can replace lost neurons and glia after spinal cord injury (SCI), and can form functional relays to re-connect spinal cord segments above and below a lesion. Previous studies grafting neural stem cells have been limited by incomplete graft survival within the spinal cord lesion cavity. Further, tracking of graft cell survival, differentiation, and process extension had not been optimized. Finally, in previous studies, cultured rat NSCs were typically reported to differentiate into glia when grafted to the injured spinal cord, rather than neurons, unless fate was driven to a specific cell type. To address these issues, we developed new methods to improve the survival, integration and differentiation of NSCs to sites of even severe SCI. NSCs were freshly isolated from embryonic day 14 spinal cord (E14) from a stable transgenic Fischer 344 rat line expressing green fluorescent protein (GFP) and were embedded into a fibrin matrix containing growth factors; this formulation aimed to retain grafted cells in the lesion cavity and support cell survival. NSCs in the fibrin/growth factor cocktail were implanted two weeks after thoracic level-3 (T3) complete spinal cord transections, thereby avoiding peak periods of inflammation. Resulting grafts completely filled the lesion cavity and differentiated into both neurons, which extended axons into the host spinal cord over remarkably long distances, and glia. Grafts of cultured human NSCs expressing GFP resulted in similar findings. Thus, methods are defined for improving neural stem cell grafting, survival and analysis of in vivo findings.     

Scaffold stiffness influences breast cancer cell invasion via EGFR-linked Mena upregulation and matrix remodeling

Clinically, increased breast tumor stiffness is associated with metastasis and poorer outcomes. Yet, in vitro studies of tumor cells in 3D scaffolds have found decreased invasion in stiffer environments. To resolve this apparent contradiction, MDA-MB-231 breast tumor spheroids were embedded in ‘low’ (2 kPa) and ‘high’ (12 kPa) stiffness 3D hydrogels comprised of methacrylated gelatin/collagen I, a material that allows for physiologically-relevant changes in stiffness while matrix density is held constant. Cells in high stiffness materials exhibited delayed invasion, but more abundant actin-enriched protrusions, compared to those in low stiffness. We find that cells in high stiffness had increased expression of Mena, an invadopodia protein associated with metastasis in breast cancer, as a result of EGFR and PLCγ1 activation. As invadopodia promote invasion through matrix remodeling, we examined matrix organization and determined that spheroids in high stiffness displayed a large fibronectin halo. Interestingly, this halo did not result from increased fibronectin production, but rather from Mena/α5 integrin dependent organization. In high stiffness environments, FN1 knockout inhibited invasion while addition of exogenous cellular fibronectin lessened the invasion delay. Analysis of fibronectin isoforms demonstrated that EDA-fibronectin promoted invasion and that clinical invasive breast cancer specimens displayed elevated EDA-fibronectin. Combined, our data support a mechanism by which breast cancer cells respond to stiffness and render the environment conducive to invasion. More broadly, these findings provide important insight on the roles of matrix stiffness, composition, and organization in promoting tumor invasion.     

Neural Stem Cells Grafts Decrease Neural Apoptosis Associated with Caspase-7 Downregulation and BDNF Upregulation in Rats Following Spinal Cord Hemisection

Transplantation of neural stem cells (NSCs) into lesioned spinal cord demonstrated a beneficial effect for neural repair, the underlying mechanism, however, remains to be elusive. Here, we showed that NSCs, possessing the capacity to differentiate toward into neurons and astrocytes, exhibit a neuroprotective effect by anti-apoptosis mechanism in spinal cord hemi-transected rats despite it did not improve behavior. Intravenous NSCs injection substantially upregulated the level of BDNF mRNA but not its receptor TrkB in hemisected spinal cord, while caspase-7, a downstream apoptosis gene of caspase-3, has been largely down-regulated. TUNEL staining showed that the number of apoptosis cells in injured spinal cord decreased significantly, compared with seen in rats with no NSCs administration. The present finding therefore provided crucial evidence to explain neuroprotective effect of NSCs grafts in hemisected spinal cord, which is associated with BDNF upregulation and caspase-7 downregulation.     

Co-transplantation of bFGF-expressing amniotic epithelial cells and neural stem cells promotes functional recovery in spinal cord-injured rats

We have previously demonstrated that amniotic epithelial cells (AECs) can enhance survival and neural differentiation of neural stem cells (NSCs) when co-cultured in basal media. In addition, the presence of basic fibroblast growth factor (bFGF) enhances this AEC function. The aim of the present study was to extend those findings and investigate whether AECs modified with the bFGF gene will also enhance NSCs survival and neural differentiation in vivo and promote repair of the injured spinal cord. Female Wistar rats were used for a contusive spinal cord injury (SCI) model. Contusive SCIs were induced using a weight-drop device at levels T9-T11. Seven days following contusion, rats received grafts of NSCs only, NSCs with AECs/pLEGFP-hbFGF, or NSCs with AECs/pLEGFP-C1 into the injured region. Significant locomotor improvement was observed in the NSCs/AECs co-graft group beginning at 3 weeks compared with the NSCs or NaCl only groups. These results were confirmed and extended in an electrophysiological analysis. An immunohistological analysis revealed that AECs/pLEGFP-hbFGF promoted the survival (vs NaCl group: 194+/-9.17 vs 103.6+/-13.05) and neural differentiation (vs NaCl group: 14.24+/-1.11 vs 7+/-0.63) of co-transplanted NSCs. We also confirmed that AECs could promote the survival of host neurons. These results suggest that AECs/pLEGFP-hbFGF improve the NSCs survival and differentiation microenvironment and may be useful as a source of sustained trophic supported to improve NSCs differentiation into neurons in vivo. These findings suggest that a cograft of AECs/pLEGFP-hbFGF and NSCs may have benefits for SCI.     

胚胎神经干细胞移植及胶质细胞源性神经营养因子对大鼠脊髓损伤的修复作用

将胚胎神经干细胞(neural stem cells,NSCs)移植至成年大鼠损伤的脊髓,观察移植后NSCs的存活、迁移以及损伤后的功能恢复。实验结果显示：动物NSCs移植4周后,斜板实验平均角度和运动评分结果比对照组均有明显增高(P＜0．05),而脊髓损伤(spinal cord injury,SCI)处的空洞面积显著减小(P＜0．05);在NSCs中加入胶质细胞源性的神经营养因子(glial cell line-derived neurotrophic factor,GDNF)后,上述改变更加显著。移植后的NSCs不仅能存活,而且向损伤的头端和尾端迁移达3mm之远。这些结果表明,移植的NSCs不仅可以存活、迁移,还可减小SCI空洞面积,促进动物神经功能的恢复;此外,我们的结果还表明GDNF对SCI功能恢复有促进作用。     

Comparison of mesenchymal stromal cells from human bone marrow and adipose tissue for the treatment of spinal cord injury

Background aimsBone marrow and subcutaneous adipose tissue are both considered prospective sources of mesenchymal stromal cells (MSCs), which can be used in cell therapy for spinal cord injury (SCI). The present study investigated whether human adipose tissue-derived mesenchymal stromal cells (hADSCs) transplanted into a rat model of SCI would lead to similar or improved neurologic effects compared with human bone marrow-derived mesenchymal stromal cells (hBMSCs).MethodshADSCs and hBMSCs were isolated from five adult donors. These MSCs were characterized using flow cytometry, immunocytochemistry, real-time polymerase chain reaction and enzyme-linked immunosorbent assay. Immediately after SCI, 2 × 10⁵ hBMSCs or hADSCs were injected into the injured spinal cord. Locomotor function, cell survival and differentiation, spinal cord tissue morphology and brain-derived neurotrophic factor (BDNF) expression were compared between groups.ResultshADSCs and hBMSCs showed similar surface protein expression, and hADSCs showed higher proliferative activity with higher expression of vascular endothelial cell growth factor, hepatocyte growth factor and BDNF than hBMSCs. After transplant, both hADSCs and hBMSCs migrated within the injured spinal cord without differentiating into glial or neuronal elements. Administration of hADSCs was associated with marked changes in the SCI environment, with significant increases in BDNF levels. This was simultaneously associated with increased angiogenesis, preserved axons, decreased numbers of ED1-positive macrophages and reduced lesion cavity formation. These changes were accompanied by improved functional recovery.ConclusionsThe present results suggest that hADSCs would be more appropriate for transplant to treat SCI than hBMSCs.