N-cadherin

摘要： 目的研究意向性运动干预治疗对大脑局部缺血大鼠行为学及梗死灶周围组织中GluA2和Ncadherin表达的影响。方法利用鱼线栓堵法将大鼠制成大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)模型共144只,随机分成模型(MCAO)组、生活环境变化(environment modification,EM)组、普通康复训练(control rehabilitstion,CR)组和意向性运动干预(willed movement,WM)组,每组根据术后时间划分为7 d、15 d、30 d 3个亚组。对各组研究对象的行为变化进行观察分析,通过免疫组化技术研究脑梗死灶周围组织中GluA2和N-cadherin蛋白水平。结果再灌7 d、15 d、30 d,意向性运动干预组的攀爬频率显著高于EM及CR组(P<0.01);再灌30 d,和MCAO组相比,意向性运动干预组的神经功能缺损评分更低(P<0.05);而其缺血半暗带区域GLUA2、N-cadherin蛋白表达量都更高(P<0.05)。结论对于大脑局部缺血所导致的神经功能障碍,意向性运动干预治疗可促进其恢复,这可能是因为该运动可激活缺血半暗带中GLUA2和N-cadherin的表达,提高突触可塑性。

摘要： 目的:分析上皮细胞间质转化(EMT)标记基因E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)与冠心病患者冠脉病变程度的相关性。方法:选取江苏省苏北人民医院心内科2020年4月—12月冠状动脉造影确诊为冠心病患者60例为研究组,基于Gensini评分将研究组分为轻度病变组(19例)、中度病变组(21例)、重度病变组(20例)。选择造影阴性30例作为对照组,均抽取外周血检测E-cadherin、N-cadherin含量,分析E-cadherin、N-cadherin与冠脉病变程度相关性。结果:研究组中重度病变组外周血清中E-cadherin含量最低,N-cadherin含量最高。对照组E-cadherin外周血清中含量最高,N-cadherin含量最低。Gensini评分与E-cadherin浓度呈负相关(r=-0.450,P<0.05),与N-cadherin浓度呈正相关(r=0.433,P<0.05)。结论:EMT相关标记基因E-cadherin、N-cadherin可能与冠心病患者冠状动脉病变程度存在相关性。

摘要： 目的探讨沉默N-cadherin对人牙髓干细胞(dental pulp stem cells,DPSCs)增殖和迁移能力的影响,为基于DPSCs的牙髓再生提供实验依据。方法采用N-cadherin shRNA慢病毒转染DPSCs,qRT-PCR、Western blot检测N-cadherin的表达水平,验证沉默效率。实验分为阴性对照组(shRNA-NC)和N-cadherin shRNA沉默组。采用CCK-8法检测细胞增殖能力,流式细胞术检测细胞周期和细胞凋亡情况,Transwell法检测细胞迁移能力。结果N-cadherin shRNA可显著降低DPSCs的N-cadherin mRNA和蛋白的表达水平(P<0.001)。N-cadherin shRNA组细胞增殖活性分别在细胞接种的第3、4天高于shRNA-NC组,第6~8天低于shRNA-NC组,差异均具有统计学意义(P<0.05);在第3天时,N-cadherin shRNA组处于S期和G2期的细胞比例大于shRNANC组(P<0.05);在第6天时,N-cadherin shRNA组细胞处于S期和G2期的比例小于shRNA-NC组(P<0.05),同时N-cadherin shRNA组凋亡细胞比例大于shRNA-NC组(P<0.01);分别以低细胞密度和高细胞密度接种于Transwell上室培养20 h,N-cadherin shRNA组穿过Transwell上室小孔的细胞数量均多于shRNA-NC组(P<0.001)。结论沉默N-cadherin表达可促进DPSCs早期的增殖和迁移能力。

摘要： 目的:探讨乳腺癌组织中上皮性钙粘蛋白(E-cadherin)表达与CD44V6、神经性钙粘蛋白(N-cadherin)表达的相关性。方法:选取2018年1月—2020年8月在我院确诊为乳腺癌的105例患者癌组织、癌旁组织和正常乳腺组织30例制作病理切片,检测各组的E-cadherin、CD44V6、N-cadherin蛋白的表达。结果:在乳腺癌组织、癌旁组织及正常乳腺组织中,E-cadherin表达在癌组织阳性率明显较低(P<0.05),CD44V6、N-cadherin表达在癌组织阳性率明显较高(P<0.05);E-cadherin表达与乳腺癌肿瘤大小、分级、TNM分期、淋巴结转移呈负相关(P<0.05),CD44V6和N-cadherin表达与乳腺癌肿瘤大小、分级、TNM分期、淋巴结转移呈正相关(P<0.05),与乳腺癌组织学类别无相关性;乳腺癌组织中E-cadherin表达与CD44V6、N-cadherin表达均呈负相关(r=-0.137,P=0.034;r=-0.213,P=0.001),CD44V6表达与N-cadherin表达呈正相关(r=0.329,P=0.000)。结论:E-Cadherin与CD44V6及N-cadherin联合检测,对于乳腺癌的发生和发展、预后的监测有着重要的意义。

摘要： 目的探讨沉默神经性钙黏附蛋白(N-cadherin)对卵巢癌细胞增殖、侵袭、迁移与上皮-间质转化的影响及分子机制。方法收集2021年3月~2021年12月我院卵巢癌患者手术切除的卵巢癌组织与邻近癌旁正常组织,采用免疫组织化学染色检测N-cadherin蛋白表达。通过RNAi技术沉默卵巢癌细胞SKOV3细胞中N-cadherin的表达后分为空白对照组、空载体组、siRNA-N-cadherin组。CCK-8法、细胞克隆形成实验、流式细胞术分别检测各组细胞增殖活性、克隆形成能力以及细胞周期分布,Transwell实验和细胞划痕实验分别检测各组细胞侵袭与迁移能力,实时荧光定量PCR检测各组细胞中N-cadherin、上皮性钙粘附分子(E-cadherin)mRNA表达,免疫细胞化学染色检测各组细胞中N-cadherin、E-cadherin、基质金属蛋白酶2(MMP2)和基质金属蛋白酶9(MMP9)蛋白表达,Western blot检测各组细胞中MEK、ERK蛋白表达。结果与癌旁正常组织比较,卵巢癌组织中N-cadherin呈高表达(P<0.05)。与空白对照组和空载体组比较,沉默N-cadherin能够抑制SKOV3细胞增殖活性(P<0.05),细胞克隆形成数目下降(P<0.01),并使细胞发生S期阻滞(P<0.01),细胞侵袭数目和迁移数目均减少(P<0.01);此外,沉默N-cadherin后SKOV3细胞中N-cadherin mRNA相对表达量下降,E-cadherin mRNA相对表达量升高(P<0.05),细胞内N-cadherin、MMP2和MMP9蛋白阳性染色减弱,E-cadherin蛋白阳性染色增强,细胞中MEK、ERK的蛋白表达水平均下降(P<0.05)。结论沉默N-cadherin可抑制卵巢癌细胞活性、细胞克隆形成、侵袭、迁移与上皮-间质转化的能力,使细胞阻滞于S期,其机制可能与阻碍MEK、ERK信号通路有关。

摘要： Asparagine-linked glycosylation protein 1 homolog(ALG1)participates in the initial stage of protein N-glycosylation and N-glycosylation has been implicated in the process of hepatocellular carcinoma(HCC)progression.However,whether ALG1 plays a role in human HCC remains unknown.In this study,the expression profile of ALG1 in tumorous and corresponding adjacent non-tumor tissues was analyzed.The relationship of ALG1 expression with clinical features and prognosis of HCC patients was also evaluated using immuno-histochemical method.Here we found ALG1 decreased in HCC tissues compared with adjacent normal liver tissues,which predicted an unfavorable prognosis.Combined with RNA interference,nascent proteome and glycoproteome were determined systematically in Huh7 cell line.Bioinformatics analysis indicated that the differentially expressed proteins participating in the response of ALG1 knockdown were most significantly associated with cell-cell adhesion.Functional studies confirmed that knockdown of ALG1 reduced cell adhesion capacity,and promoted cell migration.Furthermore,down-regulation of H8N2(on N-glycosite N651)and H5N4S2F1(on N-glycosite N692)from N-cadherin was identified as a feature of ALG1 knockdown.Our findings revealed that ALG1 controlled the expression of glycosylated N-cadherin and played a role in HCC migration,with implications for prognosis.

摘要： 目的:检测胆管细胞癌中微小RNA-194-5p(miR-194-5p)和上皮型钙黏蛋白(E-cadherin)的表达,分析其临床意义及相关性.方法:选择胆管细胞癌49例作为观察组,选择正常胆管上皮组织49例作为对照组.应用实时荧光定量PCR法(qPCR)检测两组中miR-194-5p的表达,应用免疫组化法检测观察组中E-cadherin和神经型钙黏蛋白(N-cadherin)的表达.建立胆管癌细胞系的空白对照组、空载体转染组和miR-194-5p转染组,采用Western Blot法检测E-cadherin和N-cadherin的表达.结果:胆管细胞癌中miR-194-5p的表达明显低于对照组,miR-194-5p在不同肿物最大径、神经累犯、分化程度、转移及脉管癌栓的分组中差异有统计学意义.生存分析显示miR-194-5p低表达者生存时间短.miR-194-5p与E-cadherin具有正相关性(r=0.68,P=0.014),miR-194-5p与N-cadherin具有负相关性(r=-0.62,P=0.013).与空白对照组和空载体转染组相比,miR-194-5p转染组E-cadherin的表达升高,N-cadherin的表达下降.结论:miR-194-5p在胆管细胞癌中低表达,与病变发展及生物学行为有关.miR-194-5p与E-cadherin具有正向协同作用,对调节细胞黏附有一定价值.miR-194-5p可能是胆管细胞癌独立的预后因子.

摘要： 目的研究黄精多糖对阿尔茨海默病模型斑马鱼的影响。方法将斑马鱼置于T迷宫中训练7 d,将训练成功的40条斑马鱼分为4组:空白组、模型组、阳性组的和黄精多糖组。模型组、阳性组和黄精多糖组在100μg·L-1的AlCl3中造模6 d;阳性组暴露在4μg·L-1的石杉碱甲药液,黄精多糖组暴露在6 g·L-1黄精多糖药液,均给药6 d,模型组不予以给药,空白组不做任何处理,并用录像的方法对斑马鱼各阶段进行记录,对各组在EC区的时间进行分析。给药结束后取出脑组织,采用Western印迹法测定N-cadherin,P38和p-P38蛋白因子的表达。结果在行为学上,分析在EC区所待时间,空白组、阳性组和黄精多糖组分别与模型组相比均有统计学差异(P<0.05)。在蛋白水平上,与模型组比较,阳性组和黄精多糖组P38和p-P38蛋白下调,而N-cadherin蛋白上调(P<0.05)。结论黄精多糖通过上调N-cadherin蛋白水平,阻碍P38磷酸化,改善阿尔茨海默病模型斑马鱼的学习记忆能力。

摘要： OBJECTIVE To investigate the effect of rhynchophylline on zebrafish with Alzheimer disease induced by AlCl3.METHODS Zebrafish were trained in T maze for 7 d.60 zebrafish were selected and divided into 6 groups:blank group,model group,positive group,low-dose group,medium-dose group and high-dose group,with 10 in each group.Feeding zebrafish adaptively for 2 d,starting training on the third day,and recording its movement track;Except the blank group,the other five groups were modeled and their motion trajectories were recorded.After modeling,the drug group was giv⁃en rhynchophylline,while the positive group was given huperzine A,and the movement track was recorded.The recorded movement track was analyzed by behavioral record analysis software smart 3.0.The expression levels of P38,p-P38 protein and N-cadherin protein in zebrafish brain tissue were detected by Western blotting.RESULTS The data of time spent in the green short arm area of six groups of zebrafish were compared and analyzed:there was significant dif⁃ference between blank group and model group(P<0.05),and there was significant difference between model group and positive group,low dose group,middle dose group and high dose group(P<0.05).Comparative analysis of the time percentage of zebrafish in the green arm area of six groups:there was significant differ⁃ence between the blank group and the model group(P<0.05),and there was significant differ⁃ence between the model group and the positive group,the rhynchophylline administration group,low dose group,middle dose group and high dose group(P<0.05).The protein was detected by Western blotting.Compared with the model group,the expression level of N-cadherin protein was up-regulated(P<0.05).Compared with the model group,the expression levels of p-P38 pro⁃tein in blank group,positive group and rhyncho⁃phylline administration group were down-regulat⁃ed(P<0.05).CONCLUSION Uncaria rhyncho⁃phylline can down-regulate the ratio of p-P38/P38 and up-regulate N-cadherin,thus improving the learning and memory ability of zebrafish with Alzheimer disease.

摘要： 目的 探索上皮-间质转化(epithelial mesenchymal transition,EMT)与慢性胰腺炎(chronic pancreatitis,CP)胰腺纤维化的关系及大柴胡汤防治CP的机制.方法 健康昆明小鼠78只,随机分为:空白组(n=18)、CP组(n=30)及大柴胡汤组(n=30).CP组和大柴胡汤组腹腔注射20％左旋精氨酸(L-arginine,3 g/kg,每周1次),空白组腹腔注射等体积的生理盐水.大柴胡汤组在注射L-arginine 1周后,同时给予注射L-arginine和大柴胡汤灌胃治疗5周(14 g/kg,每日一次).从给药开始计时,各组分别于第2,4,6周麻醉处死小鼠,摘取小鼠胰腺组织,通过HE染色观察胰腺组织的病理改变,免疫组化观察胰腺组织中E-cadherin、N-cadherin、α-SMA的表达变化,Western blotting检测胰腺组织中E-cadherin、N-cadherin蛋白含量的改变.结果 与空白组相比,CP组小鼠胰腺组织中胶原纤维沉积随着造模时间延长不断增加,α-SMA表达逐渐增多,E-cadherin表达逐渐减少,在第4,6周时最为显著(P<0.01),而N-cadherin表达逐渐增多,在第6周最为显著(P<0.01).与CP组相比,大柴胡汤组第4,6周仅见部分腺泡萎缩及少量炎细胞浸润,胶原纤维沉积明显减轻,α-SMA表达显著减少,E-cadherin在第6周显著升高(P<0.01),而N-cadherin在第6周显著下降(P<0.01).结论 在20％L-arginine刺激作用下,小鼠胰腺实质细胞转化为间质细胞,表明EMT参与CP胰腺纤维化的进展.大柴胡汤可通过抑制EMT转化,减轻胰腺纤维化,对CP有明显治疗作用.