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laminin-coated multifilament entubulation, combined with Schwann cells and glial cell line-derived neurotrophic factor, promotes unidirectional axonal regeneration in a rat model of thoracic spinal cord hemisection

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Neural Regeneration Research 2021年第1期16卷 186-191页

作者： Ling-Xiao Deng Nai-Kui Liu Ryan Ning Wen Shuang-Ni Yang Xuejun Wen Xiao-Ming Xu Spinal Cord and Brain Injury Research Group Stark Neurosciences Research InstituteIndiana University School of MedicineIndianapolisINUSA Department of Neurological Surgery Indiana University School of MedicineIndianapolisINUSA Maggie L.Walker Governor’s School RichmondVAUSA Institute for Engineering and Medicine Department of Chemical and Life Science EngineeringVirginia Commonwealth UniversityRichmondVAUSA

Biomaterial bridging provides physical substrates to guide axonal growth across the lesion.To achieve efficient directional guidance,combinatory strategies using permissive matrix,cells and trophic factors are necessary.In the present study,we evaluated permissive effect of poly(acrylonitrile-co-vinyl chloride)guidance channels filled by different densities of laminin-precoated unidirectional polypropylene filaments combined with Schwann cells,and glial cell line-derived neurotrophic factor for axonal regeneration through a T10 hemisected spinal cord gap in adult rats.We found that channels with filaments significantly reduced the lesion cavity,astrocytic gliosis,and inflammatory responses at the graft-host boundaries.The laminin coated low density filament provided the most favorable directional guidance for axonal regeneration which was enhanced by co-grafting of Schwann cells and glial cell line-derived neurotrophic factor.These results demonstrate that the combinatorial strategy of filament-filled guiding scaffold,adhesive molecular laminin,Schwann cells,and glial cell line-derived neurotrophic factor,provides optimal topographical cues in stimulating directional axonal regeneration following spinal cord injury.This study was approved by Indiana University Institutional Animal Care and Use Committees(IACUC#:11011)on October 29,2015.

关键词： axonal regeneration extracellular molecule filament density hemisection laminin neurotrophic factor Schwann cell spinal cord injury thoracic transplantation

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The importance of laminin at the blood-brain barrier

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Neural Regeneration Research 2023年第12期18卷 2557-2563页

作者： Sebok K.Halder Arjun Sapkota Richard Milner San Diego Biomedical Research Institute San DiegoCAUSA

The blood-brain barrier is a unique property of central nervous system blood vessels that protects sensitive central nervous system cells from potentially harmful blood components.The mechanistic basis of this barrier is found at multiple levels,including the adherens and tight junction proteins that tightly bind adjacent endothelial cells and the influence of neighboring pericytes,microglia,and astrocyte endfeet.In addition,extracellular matrix components of the vascular basement membrane play a critical role in establishing and maintaining blood-brain barrier integrity,not only by providing an adhesive substrate for blood-brain barrier cells to adhere to,but also by providing guidance cues that strongly influence vascular cell behavior.The extracellular matrix protein laminin is one of the most abundant components of the basement membrane,and several lines of evidence suggest that it plays a key role in directing blood-brain barrier behavior.In this review,we describe the basic structure of laminin and its receptors,the expression patterns of these molecules in central nervous system blood vessels and how they are altered in disease states,and most importantly,how genetic deletion of different laminin isoforms or their receptors reveals the contribution of these molecules to blood-brain barrier function and integrity.Finally,we discuss some of the important unanswered questions in the field and provide a“to-do”list of some of the critical outstanding experiments.

关键词： astrocytes basement membrane blood vessels blood-brain barrier integrity dystroglycan endothelial cells inflammation integrins laminin pericytes

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p53 positively regulates the proliferation of hepatic progenitor cells promoted by laminin-521

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Signal Transduction and Targeted Therapy 2022年第9期7卷 3592-3608页

作者： Mingyang Ma Shuyao Hua Xiangde Min Liang Wang Jun Li Ping Wu Huifang Liang Bixiang Zhang Xiaoping Chen Shuai Xiang Department of General Surgery Tianjin Medical University General HospitalTianjinChina Hepatic Surgery Center Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina Department of Clinical Nutrition Shanghai Jiao Tong University Affliated Sixth People's HospitalShanghaiChina Department of Radiology Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina Wuhan Pulmonary Hospital Wuhan Institute for Tuberculosis ControlWuhanChina Department of Pathophysiology Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina Hubei Key Laboratory of Hepato-Pancreato-Biliary Disease WuhanChina Key Laboratory of Organ Transplantation Ministry of EducationWuhanChina Key Laboratory of Organ Transplantation National Health CommissionWuhanChina Key Laboratory of Organ Transplantation Chinese Academy of Medical SciencesWuhanChina

Hepatic progenitor cells(HPCs)hold tremendous potential for liver regeneration,but their well-known limitation of proliferation hampers their broader use.There is evidence that laminin is required for the proliferation of HPCs,but the laminin isoform that plays the dominant role and the key intracellular downstream targets that mediate the regulation of HPC proliferation have yet to be determined.Here we showed that p53 expression increased gradually and reached maximal levels around 8 days when lamininα4,α5,β2,β1,andγ1 subunit levels also reached a maximum during HPC activation and expansion.laminin-521(LN-521)promoted greater proliferation of HPCs than do laminin,matrigel or other laminin isoforms.Inactivation of p53 by PFT-αor Ad-p53^(V143A) inhibited the promotion of proliferation by LN-521.Further complementary MRI and bioluminescence imaging analysis showed that p53 inactivation decreased the proliferation of transplanted HPCs in vivo.p53 was activated by LN-521 through the Integrinα6β1/FAK-Src-Paxillin/Akt axis.Activated p53 was involved in the nuclear translocation of CDK4 and inactivation of Rb by inducing p27^(Kip1).Taken together,this study identifies LN-521 as an ideal candidate substrate for HPC culture and uncovers an unexpected positive role for p53 in regulating proliferation of HPCs,which makes it a potential target for HPC-based regenerative medicine.

关键词： laminin progenitor inhibited

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Microenvironmental stiffness mediates cytoskeleton re-organization in chondrocytes through laminin-FAK mechanotransduction

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International Journal of Oral Science 2022年第2期14卷 182-191页

作者： Chenchen Zhou Mengmeng Duan Daimo Guo Xinmei Du Demao Zhang Jing Xie State Key Laboratory of Oral Diseases&National Clinical Research Center for Oral Diseases West China Hospital of StomatologySichuan UniversityChengduChina Department of Cariology and Endodontics West China Hospital of StomatologySichuan UniversityChengduChina

Microenvironmental biophysical factors play a fundamental role in controlling cell behaviors including cell morphology,proliferation,adhesion and differentiation,and even determining the cell fate.Cells are able to actively sense the surrounding mechanical microenvironment and change their cellular morphology to adapt to it.Although cell morphological changes have been considered to be the first and most important step in the interaction between cells and their mechanical microenvironment,their regulatory network is not completely clear.In the current study,we generated silicon-based elastomer polydimethylsiloxane(PDMS)substrates with stiff(15:1,PDMS elastomer vs.curing agent)and soft(45:1)stiffnesses,which showed the Young’s moduli of~450 k Pa and 46 kPa,respectively,and elucidated a new path in cytoskeleton re-organization in chondrocytes in response to changed substrate stiffnesses by characterizing the axis shift from the secreted extracellular protein lamininβ1,focal adhesion complex protein FAK to microfilament bundling.We first showed the cellular cytoskeleton changes in chondrocytes by characterizing the cell spreading area and cellular synapses.We then found the changes of secreted extracellular linkage protein,lamininβ1,and focal adhesion complex protein,FAK,in chondrocytes in response to different substrate stiffnesses.These two proteins were shown to be directly interacted by Co-IP and colocalization.We next showed that impact of FAK on the cytoskeleton organization by showing the changes of microfilament bundles and found the potential intermediate regulators.Taking together,this modulation axis of lamininβ1-FAK-microfilament could enlarge our understanding about the interdependence among mechanosensing,mechanotransduction,and cytoskeleton re-organization.

关键词： laminin mec stiffness

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Effects of Acupuncture at Baihui(GV 20) and Zusanli(ST 36) on Peripheral Serum Expression of MicroRNA 124,laminin and Integrin β1 in Rats with Cerebral Ischemia Reperfusion Injury

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Chinese Journal of Integrative Medicine 2016年第1期22卷 49-55页

作者：陈素辉孙华张亚敏徐虹杨杨王富明 Department of Traditional Chinese Medicine Peking Union Medical College Hospital Peking Union Medical CollegeChinese Academy of Medical Sciences

Objective： To explore the effects of acupuncture at Baihui （GV 20） and Zusanli （ST 36） on the peripheral serum expression of microRNA 124 （miRNA 124）, laminin and integrinβ 1 in rats with cerebral ischemia reperfusion injury （CIRI）. Methods： Seventy-two healthy male Sprague-Dawley rats were randomized into a model group, an acupuncture group, and a sham-operated group using a random digits table, with 24 rats per group. Each group was further randomly divided into 1-, 3-, 5-, and 7-day subgroups based on the reperfusion time according to a random digits table, with 6 rats in each subgroup. In the model and acupuncture groups, CIRI was induced using the thread occlusion method. Electroacupuncture stimulation was applied daily to GV 20 and left ST 36 for 20 min at the indicated time points after successful operations. Serum was sampled for detecting laminin and integrin β 1 protein via enzyme-linked immunosorbent assay, and serum miRNA 124 was examined using quantitative polymerase chain reaction. Results： The serum level of miRNA 124 in the cerebral ischemia rats increased significantly, and the peak expression of miRNA 124 in both the model and acupuncture groups occurred at 3 days. The expression of miRNA 124 in the acupuncture group was higher than in the model group at the same time point （5.96±0.01 vs. 3.11 ±0.04, P〈0.05）. laminin expression in serum from the cerebral ischemia group was higher than that in the sham-operated group. Compared with the model group, the level of laminin in the serum of the acupuncture group was significantly lower at each time point, especially at the 3-day, and 7-day time points （589.12± 3.57 vs. 793.05± 5.28, and 600.53± 3.05 vs. 899.06± 5.74, P〈0.05）. The level of integrin 131 in the serum from the acupuncture group was lower than that in the model group particularly at the 3-day and Z-day time points （208.66± 0.95 vs. 280.83± 1.77, and 212.36±0.95 vs. 316.77±2.42, P〈0.05）. Additionally, the model group and the acupuncture group showed dual peaks of integrin β 1 and laminin exp

关键词： acupuncture cerebral ischemia reperfusion injury microRNA 124 laminin integrin β 1

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Inhibition on Apoptosis Induced by Elevated Hydrostatic Pressure in Retinal Ganglion Cell-5 via laminin Upregulating β1-integrin/Focal Adhesion Kinase/Protein Kinase B Signaling Pathway

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Chinese Medical Journal 2016年第8期129卷 976-983页

作者： Yi Li Yan-Ming Chen Ming-Ming Sun Xiao-Dan Guo Ya-Chen Wang Zhong-Zhi Zhang Department of Ophthalmology First Affiliated Hospital China Medical University Shenyang Liaoning 110001 China

Background：Glaucoma is a progressive optic neuropathy characterized by degeneration of neurons due to loss of retinal ganglion cells （RGCs）.High intraocular pressure （HIOP）,the main risk factor,causes the optic nerve damage.However,the precise mechanism of HIOP-induced RGC death is not yet completely understood.This study was conducted to determine apoptosis of RGC-5 cells induced by elevated hydrostatic pressures,explore whether laminin is associated with apoptosis under pressure,whether laminin can protect RGCs from apoptosis and affirm the mechanism that regulates the process of RGCs survival.Methods：RGC-5 cells were exposed to 0,20,40,and 60 mmHg in a pressurized incubator for 6,12,and 24 h,respectively.The effect of elevated hydrostatic pressure on RGC-5 cells was measured by Annexin V-fluorescein isothiocyanate/propidium iodide staining,3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay,and Western blotting of cleaved caspase-3 protein.Location and expression oflaminin were detected by immunofluorescence.The expression of β 1-integrin,phosphorylation of focal adhesion kinase （FAK） and protein kinase B （PKB,or AKT） were investigated with real-time polymerase chain reaction and Western blotting analysis.Results：Elevated hydrostatic pressure induced apoptosis in cultured RGC-5 cells.Pressure with 40 mmHg for 24 h induced a maximum apoptosis.laminin was declined in RGC-5 cells after exposing to 40 mmHg for 24 h.After pretreating with laminin,RGC-5 cells survived from elevated pressure.Furthermore,β1-integrin and phosphorylation of FAK and AKT were increased compared to 40 mmHg group.Conclusions：The data show apoptosis tendency of RGC-5 cells with elevated hydrostatic pressure.laminin can protect RGC-5 cells against high pressure via β 1-integrin/FAK/AKT signaling pathway.These results suggest that the decreased laminin of RGC-5 cells might be responsible for apoptosis induced by elevated hydrostatic pressure,and laminin or activating β1-integrin/FAK/AKT pathway might be potential treatments to p

关键词： Apoptosis Glaucoma Hydrostatic Pressure laminin Retinal Ganglion Cells

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laminin在成骨及成软骨分化BMSCs脱细胞片层中的差异性研究

Laminin在成骨及成软骨分化BMSCs脱细胞片层中的差异性研究

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2020年中华口腔医学会口腔生物医学专业委员会第十次全...

作者：陈朝真姜治伟冯煜婷杨国利浙江大学附属口腔医院

目的:细胞外基质(Extracellular matrix, ECM)为干细胞提供了一个合适的三维环境,即细胞龛。细胞可以在细胞龛中进行增殖、粘附、再生和分化。间充质干细胞的ECM在脱细胞后产生的基质能够极大程度上消除ECM的免疫原性,并保持引导干细胞... 详细信息

目的:细胞外基质(Extracellular matrix, ECM)为干细胞提供了一个合适的三维环境,即细胞龛。细胞可以在细胞龛中进行增殖、粘附、再生和分化。间充质干细胞的ECM在脱细胞后产生的基质能够极大程度上消除ECM的免疫原性,并保持引导干细胞分化的功能。层粘连蛋白(laminin, LN)作为ECM的重要组成成分,可在体内外调节细胞表型和分化。但是,到目前为止,尚无研究探讨laminin在不同类型ECM中的表达差异。本研究致力于通过蛋白质组学分析来探究laminin在成骨诱导后、成软骨诱导后及正常培养液培养后的BMSCs脱细胞片层中的差异表达情况。方法:本实验提取了大鼠骨髓间充质干细胞(Bone marrow mesenchymal stem cells, BMSCs),用成骨诱导液、成软骨诱导液和普通培养基培养7天,脱细胞后形成BMSCs脱细胞片层。通过扫描电镜、透射电镜和X射线光电子能谱来确认脱细胞是否成功;采用基于质谱的蛋白质组学分析方法筛选出成骨分化BMSCs脱细胞片层(成骨组),成软骨分化BMSCs脱细胞片层(成软骨组)和正常培养基培养的BMSCs脱细胞片层(对照组)中显著差异表达的laminin相关蛋白;通过聚类分析、热图分析、GO富集分析、KEGG富集分析等方法对数据进行分析。结果:扫描电镜、透射电镜图像和和X射线光电子能谱数据证明成骨组、成软骨组和对照组三组BMSCs片层脱细胞成功。比较成骨组、成软骨组和对照组,热图分析显示共有24个与层粘连蛋白相关的蛋白变化程度超过1.2倍。通过聚类分析结果显示,与对照组相比,成软骨组中LN-α3,LN-α4,LN-α5,LN-β1,LN-β2和LN-γ1的表达含量明显减少。对比成骨组与对照组,LN-α3和LN-α4显著下调,而LN-α5显著上调。此外,网络分析发现laminin会与整合素及其他细胞表面受体分子产生相互作用。比较成骨组与对照组发现,laminin主要参与了64条蛋白质-蛋白质相互作用网络,对比成软骨组与对照组后鉴定出了54条laminin相关的蛋白质-蛋白质相互作用网络。结论:laminin在成骨诱导后、成软骨诱导后及正常培养液培养后的BMSCs脱细胞片层中蛋白表达存在差异。laminin的差异性分析将为其调控BMSCs成骨分化和成软骨分化提供新的思路及策略。

关键词： laminin BMSCs脱细胞片层成骨分化成软骨分化蛋白质组学

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laminin蛋白对UBM上BMSCs增殖及其向平滑肌细胞分化的影响

Laminin蛋白对UBM上BMSCs增殖及其向平滑肌细胞分化的影响

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作者：魏桂枝第三军医大学

学位级别：硕士

研究背景与目的压力性尿失禁(Stress urinary incontinence,SUI)是老年妇女的常见临床疾病,在老年妇女中的发病率高达27.6%,严重影响了广大妇女生活质量[1-3]。SUI的发病机制与女性盆底结构改变密切相关,目前已经得到妇产科学界的重视... 详细信息

研究背景与目的压力性尿失禁(Stress urinary incontinence,SUI)是老年妇女的常见临床疾病,在老年妇女中的发病率高达27.6%,严重影响了广大妇女生活质量[1-3]。SUI的发病机制与女性盆底结构改变密切相关,目前已经得到妇产科学界的重视。手术治疗是SUI的常用治疗方法,悬吊术已成为SUI治疗的一线手术方式[4,5]。目前临床运用的吊带材料包括合成材料和生物材料[6]。合成材料因其具有组织侵蚀性强、瘢痕形成等难以克服的缺陷,不利于临床广泛推广。而经脱细胞处理的生物材料因保留了组织的完整性,且免疫源性低,已被广泛应用于组织工程和再生医学[7],脱细胞膀胱基质(Urinary bladder matrix,UBM)是生物材料的一种,初步研究表明其在SUI的治疗中具有一定的潜能[8],但面临的最大问题是降解周期短,从而导致SUI复发。一些学者提出将干细胞种植于生物材料上进行替代治疗可能是SUI的有效治疗手段[9],然而却存在着负载骨髓间充质干细胞(Bone marrow mesenchymal stem cells,BMSCs)的UBM存在着BMSCs增殖率低以及向平滑肌细胞分化程度低的缺点[10],因此,如何提高UBM上BMSCs的增殖率及向平滑肌细胞分化率是目前亟待解决的热点问题。已有研究报道,laminin蛋白通过其LG端功能域与BMSCs表面的整合素受体相结合从而促进BMSCs的增殖[11],并且有报道称将laminin蛋白加入到培养人的骨髓间充质干细胞培养基中,作为体外扩增人的骨髓间充质干细胞的一种常用方法[12]。同时,laminin蛋白可促进BMSCs向成骨细胞、骨细胞等方向分化[13]。而laminin蛋白不仅是构成UBM材料基底层的基本结构,而且是UBM基底层最重要的功能蛋白[14]。但是UBM上的laminin蛋白是否具有促进负载BMSCs增殖及向平滑肌细胞分化的功能尚不明确。结合我们的前期研究发现经过脱细胞处理后的UBM上laminin蛋白的LG域在基底层面处于向外的游离状态,而通过交联剂交联后可使部分LG域相互融合而丧失功能[15],由此进一步证明laminin蛋白的LG功能域在BMSCs增殖、迁移以及分化中的作用。因此,我们推测ubm上的laminin蛋白也具有促进bmscs增殖及向平滑肌细胞分化的功能,探讨ubm上laminin蛋白的这一功能,对现有的ubm进行改善,从而为sui新型生物材料的构建奠定基础。1实验方法1.1bmscs的制备采用全骨髓贴壁法分离、培养3-4周雌性sd大鼠的bmscs,运用流式细胞学技术检测bmscs的免疫表型,进行干细胞鉴定,将p3代bmscs作为种子细胞。1.2ubm的制备采用机械分离和酶消化等方法对新鲜猪膀胱进行处理,达到脱细胞目的,进行冷冻干燥、消毒灭菌后密封保存,作为组织工程材料。1.3ubm复合材料的制备采用蛋白孵育法,制备ubm+laminin材料以及ubm+anti-laminin材料,进行免疫组化学检测laminin蛋白在复合材料组及单纯ubm组的表达情况。1.***.ldh检测bmscs活性取p3代bmscs,按3000/孔接种于96孔板,实验组为ubm+laminin组、ubm组和ubm+anti-laminin材料,每孔加120μl的细胞培养液,每组设置5个复孔,置于37℃,5%co2及饱和湿度下培养,隔日换液,并观察培养液有无浑浊。分别取培养1d、3d、5d、7d、9d细胞上清液行ldh检测。1.5ckk8检测bmscs增殖实验组分为3组,分别为:ubm+laminin组、ubm组和ubm+anti-laminin组。每组5个附孔,每孔3000个bmscs接种于96孔板,于培养1、3、5、7、9、12、14天的组织行cck8检测。1.6he染色观察bmscs迁移情况将p3代bmscs(1×105/cm2)接种于三组材料上,定期观察培养液有无浑浊,于接种3d、5d、7d、14d的组织行he染色,定向观察bmscs在不同材料上的迁移情况。1.7免疫组化学技术检测bmscs向smc分化情况取接种第14天的组织行免疫学检测bmscs表达α-sma情况,观察bmscs向平滑肌细胞分化情况。1.8统计学分析统计学分析应用spss17.0统计软件,计量资料以x±s表示,组间比较采用单因素方差分析。2实验结果2.1bmscs的分离、培养及鉴定显微镜下观察原代分离的bmscs呈漩涡状、贴壁生长,杂细胞较多,培养1周后细胞融合达90%。传代后细胞较均一,形态规则,呈长梭型,增殖快。流式细胞学技术检测bmscs表达中胚层来源分子cd29,阳性率为98.0%,表达内皮细胞特征分子cd31,阳性率为4.56%,表达造血干细胞系特征分子cd34,阳性率为3.30%,符合bmscs的特征。2.2ubm的制备肉眼观察ubm材料呈致密均匀的半透明的膜状组织,

关键词：膀胱猪脱细胞基质骨髓间充质干细胞 laminin 增殖迁移分化

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高渗氯化钠羟乙基淀粉40对大鼠脑缺血再灌注损伤后MMP-9和层粘连蛋白表达的影响

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中风与神经疾病杂志 2017年第1期34卷 9-12页

作者：刘星芳张秀梅赖丽娟舒鹏黄志华占丽芳单热爱黄诚赣南医学院研究生学院江西赣州341000 赣南医学院江西赣州341000 赣南医学院生理学教研室江西赣州341000 赣南医学院疼痛医学研究所江西赣州341000 赣南医学院第一附属医院麻醉科江西赣州341000

目的探讨高渗氯化钠羟乙基淀粉40(HSH)对大鼠脑缺血-再灌注(I/R)损伤后神经功能行为学评分,脑水肿情况,脑梗死体积以及脑组织中基质金属蛋白酶-9(MMP-9)和层粘连蛋白(laminin)表达的影响。方法48只雄性SD大鼠随机分为假手术组、模型组... 详细信息

目的探讨高渗氯化钠羟乙基淀粉40(HSH)对大鼠脑缺血-再灌注(I/R)损伤后神经功能行为学评分,脑水肿情况,脑梗死体积以及脑组织中基质金属蛋白酶-9(MMP-9)和层粘连蛋白(laminin)表达的影响。方法48只雄性SD大鼠随机分为假手术组、模型组、实验1组、实验2组。大鼠局灶性脑I/R损伤模型制备成功后120 min,假手术组和模型组静脉输注0.9%生理盐水,实验1组和2组分别静脉输注HSH 4 ml/kg和8 ml/kg。再灌注24 h后,行神经功能行为学评分,观察脑水肿情况,TTC染色测算脑梗死体积,用免疫印迹方法检测缺血区脑组织MMP-9和laminin的表达。结果与假手术组比较,模型组神经功能行为学评分升高(Plaminin蛋白表达降低(Plaminin蛋白表达升高(Plaminin蛋白表达均无明显变化。结论用HSH治疗大鼠脑I/R损伤后,大鼠神经功能行为学评分改善,脑水肿减轻,脑梗死体积减小,血脑屏障通透性降低,其机制可能与降低MMP-9和增加laminin蛋白的表达有关。且保护作用以4 ml/kg剂量效果更佳。

关键词：脑缺血再灌注损伤高渗氯化钠羟乙基淀粉40 MMP-9 laminin

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不同空间排列静电纺丝对RSC96细胞表达NGF及laminin的研究

引用

中国临床解剖学杂志 2015年第1期33卷 60-63页

作者：曹欣赵利张贤祚秦建强李红吴刚南方医科大学附属小榄医院神经内科广东中山528415 南方医科大学解剖学教研室广东省组织构建与检测重点实验室广州510515 澳门科技大学中医药学院澳门999078 暨南大学理工学院材料科学与工程系广州510632 华南理工大学广州510641

目的研究不同空间排列静电纺丝对雪旺细胞株(RSC96)神经营养因子(NGF)及层粘连蛋白(laminin,LN)的表达情况。方法根据培养条件不同分3组,即平行静电纺丝培养组(A)、随机静电纺丝培养组(B)及膜上培养组(C)培养3天后用Trizol法提取细胞总R... 详细信息

目的研究不同空间排列静电纺丝对雪旺细胞株(RSC96)神经营养因子(NGF)及层粘连蛋白(laminin,LN)的表达情况。方法根据培养条件不同分3组,即平行静电纺丝培养组(A)、随机静电纺丝培养组(B)及膜上培养组(C)培养3天后用Trizol法提取细胞总RNA,用Oligo(d T)逆转录酶得到c DNA。经特定引物PCR扩增,取相同量的逆转录产物进行q PCR反应,检测NGF及LN的表达差异。结果平行静电纺丝组,随机静电纺丝组,膜上培养组上NGF基因表达依次递增,各组间差异明显;LN基因的表达依次递减,各组间差异明显。结论不同空间排列的静电纺丝对RSC96细胞表达NGF和laminin影响不同,平行的静电纺丝有促进细胞向成熟分化的可能。

关键词： RSC96细胞静电纺丝 NGF laminin

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