受体,LDL

摘要： 戊二酸尿症Ⅱ型是一种影响脂肪酸、氨基酸和胆碱代谢的常染色体隐性遗传病,是引起脂质沉积病的重要病因.该病儿童少见,临床医生认识不足,易误诊、漏诊.伴腹痛、脊髓损伤的晚发型戊二酸尿症Ⅱ型更为罕见.本文对1例伴腹痛、脊髓损伤的晚发型戊二酸尿症Ⅱ型合并家族性高胆固醇血症的患儿进行回顾性分析.早期进行血脂、尿有机酸和基因检测对患儿的诊治及提高生活质量有重要意义.

摘要： Objective To explore the potential mechanisms of low density lipoprotein receptor (LDLr) in high glucose peritoneal dialysis solution (PDS)-induced peritoneal fibrosis.Methods Human peritoneal mesothelial cells (PMCs) were applied.In pre-experiment,human PMCs were cultured with 1.5％ PDS,2.5％ PDS and 4.25％ PDS for 6 h,12 h and 24 h.4.25％ mannitol was used as high osmotic pressure control.In formal experiment,PMCs were divided into the control group (treated with phosphate buffer saline) and the high glucose PDS group (treated with 4.25％ PDS for 24 h).Morphological change of PMCs was observed by inverted microscope.The mRNA and protein expressions of extracellular matrix proteins such as α-smooth muscle actin (α-SMA),fibroblast specific protein-1 (FSP-1) and collagen Ⅰ in PMCs were respectively measured by real-time PCR and Western blotting.The lipid accumulation was observed by oil red O staining and filipin staining,and the content of intracellular cholesterol ester was detected by high-performance liquid chromatography.The co-expression of the sterol regulatory element-binding protein (SREBP) cleavage-activating protein (SCAP) with golgin was observed with immunofluorescent staining.The mRNA and protein expressions of LDLr,SREBP-2 and SCAP were respectively detected by real-time PCR and Western blotting.The mRNA and protein expressions of mammalian target of rapamycin (mTOR),eukaryotic initiation factor 4E-binding protein 1 (4EBP1),and p70 S6 kinase (S6K1) were respectively detected by real-time PCR and Western blotting.Results (1) Compared with the 1.50％ PDS stimulation,4.25％ PDS for 24 h intervention significantly increased the expression of LDLr in PMCs (P ＜ 0.05),and high osmotic pressure control at 6 h,12 h and 24 h had no statistical difference (P ＞ 0.05).(2) Compared with those in the control group,in high glucose PDS group PMCs showed notable elongation consistent with the morphology of myofibroblasts,the expressions of α-SMA,FSP-1 and collagen Ⅰ were increased (all P ＜ 0.05),and the intracellular cholesterol were enhanced (P ＜ 0.05).Meanwhile,the co-expression of SCAP with golgin was enhanced,and the mRNA and protein expressions of LDLr,SREBP-2 and SCAP were up-regulated in high glucose PDS group (all P ＜ 0.05).Further,the mRNA and protein phosphorylation of mTOR,4EBP1 and S6K1 were increased (all P ＜ 0.05).Conclusions The disruption of LDLr feedback regulation is involved in high glucose PDS-mediated cholesterol accumulation in PMCs by mammalian target of rapamycin complex 1 (mTORC1) pathway,which promotes the accumulation of extracellular matrix and peritoneal fibrosis.%目的 观察低密度脂蛋白受体(LDLr)表达失调在高糖腹膜透析液(PDS)诱导腹膜纤维化中的作用及可能机制.方法 以人腹膜间皮细胞(PMC)为研究对象.分别在含1.50％、2.50％及4.25％葡萄糖的PDS中培养PMC 6、12、24 h以筛选高糖PDS组的干扰条件.以4.25％甘露醇作为高渗对照.细胞分为对照组(PBS干预)和高糖PDS组(含4.25％葡萄糖的PDS干预24 h).倒置显微镜观察细胞形态变化;实时定量PCR和Western印迹分别检测平滑肌肌动蛋白0(α-SMA)、成纤维细胞特异性蛋白1(FSP-1)、胶原Ⅰ的mRNA和蛋白表达;油红0染色及Filipin染色观察PMC中脂质沉积情况,高效液相色谱测定细胞内胆固醇酯含量;免疫荧光双重染色观察胆固醇调节元件结合蛋白(SREBP)裂解激活蛋白(SCAP)与高尔基体共表达情况,实时定量PCR和Western印迹分别检测LDLr、SREBP-2及SCAP的mRNA和蛋白表达;实时定量PCR和Western印迹分别检测雷帕霉素靶蛋白(mTOR)、真核细胞翻译起始因子4E结合蛋白1(4EBP1)、效应子核糖体蛋白S6激酶1(S6K1)的mRNA和蛋白表达.结果 (1)与1.50％PDS干预组比较,4.25％PDS刺激PMC 24 h后,细胞内LDLr表达明显升高(P＜0.05);各时间点高渗对照组细胞内LDLr表达无明显改变(P＞0.05).(2)与对照组比较,高糖PDS组细胞形态由圆形铺路石样向长梭形转变,而且胞外基质蛋白α-SMA、FSP-1及胶原Ⅰ的表达均增加(均P＜ 0.05),细胞内脂质沉积增多(P＜0.05),细胞内SCAP与高尔基的共表达明显增多,LDLr、SREBP-2及SCAP的mRNA和蛋白表达随之升高(均P＜0.05),mTOR、4EBP1及S6K1的mRNA和磷酸化蛋白表达升高(均P＜0.05).结论 高糖PDS可能通过激活PMC细胞内雷帕霉素靶蛋白复合物1(mTORC1)通路,破坏LDLr负反馈调节,从而导致细胞内脂质沉积增加,胞外基质积聚,促进腹膜纤维化发生.

摘要： Objective To investigate the protective mechanism of MEK1/2 inhibitor PD98059 on ox-LDL induced injury of human umbilical vein endothelial cells (HUVEC),and its influence on the expression of LOX-1.Methods HUVEC damage models were established by using ox-LDL and were treated with PD98059 later,divided into the negative control group,the ox-LDL group,the positive control group and the PD98059+ox-LDL group.The effect of inhibition of MEK1/2 on ox-LDL induced HUVEC damage was measured.Results Compared with the negative control group,the levels in the ox-LDL group of LOX-1,pMEK1/2,RhoA,ROCK1,ROCK2,TNF-α and IL-6 were increased significantly,the proliferations of HUVEC and the productions of NO were decreased (P＜0.05).Compared with the ox-LDL group,the levels in the positive control group and the PD98059+ox-LDL group of pMEK1/2,RhoA,ROCK1,ROCK2,TNF-α and IL-6 were decreased,the proliferation of HUVEC and the production of NO were increased (P＜0.05).Conclusion PD98059 inhibit the MEK1/2 signaling pathway to suppress the ox-LDL induced damage of HUVEC by decreasing the expression of LOX-1.%目的 探讨MEK1/2抑制剂PD98059对氧化型低密度脂蛋白(ox-LDL)诱导人脐静脉内皮细胞(HUVEC)损伤的保护机制及对低密度脂蛋白受体-1(LOX-1)表达的影响.方法 使用ox-LDL诱导HUVEC损伤模型并进行PD98059处理,分阴性对照组、ox-LDL组、阳性对照组和PD98059+OX-LDL组;检测抑制MEK1/2对ox-LDL诱导HUVEC损伤的影响.结果 与阴性对照组比较,ox-LDL组LOX-1、pMEK1/2、RhoA、ROCK1、ROCK2、肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的释放增多,细胞增殖能力及培养液中一氧化氮(NO)水平降低(P＜0.05);与ox-LDL组比较,阳性对照组和PD98059+ox-LDL组LOX-1、pMEK1/2、RhoA、ROCK1、ROCK2、TNF-α和IL-6的释放减少,细胞增殖能力及NO水平增高(P＜0.05).结论 PD98059抑制MEK1/2信号通路可降低HUVEC中LOX-1的表达以减轻ox-LDL诱导的内皮细胞损伤.

摘要： 目的 探讨低密度脂蛋白受体(low density lipoprotein receptor,LDLR)纯合子突变高胆固醇血症大鼠脑白质组织病理学改变与野生型大鼠的差异.方法 选取30只LDLR-/-大鼠和28只野生型大鼠,分别在15、l8和26周龄时采用酶联免疫法测定血浆胆固醇水平,通过透射电子显微镜观察脑胼胝体区域轴索结构,通过蛋白质印迹法对胼胝体区髓鞘碱性蛋白(myelin basic protein,MBP)进行定量分析.在26周龄时通过快蓝染色法染髓鞘,免疫荧光染色法进一步检测脑白质区MBP表达水平并观察神经胶质细胞形态学改变.结果 与野生型大鼠相比,LDLR-/-大鼠血浆胆固醇浓度明显升高,26周时可高达3.3倍.电镜结果显示,LDLR-/-大鼠自15周出现轴索损伤,且随时间进展逐渐加重.26周时,LDLR-/-大鼠蛋白质印迹分析显示胼胝体区MBP表达水平显著降低,快蓝染色显示胼胝体区神经纤维出现疏松、弥散性空泡形成及有髓神经纤维缺失.此外,还发现LD LR--大鼠少突胶质细胞数量明显减少,星形胶质细胞及小胶质细胞大量激活.结论 LDLR-/-大鼠会出现自发性高胆固醇血症,在成年早期脑白质区即存在轴索损伤、髓鞘脱失、少突胶质细胞减少以及星形胶质细胞和小胶质细胞异常激活.%Objective To investigate the difference between histopathological changes of brain white matter in low-density lipoprotein receptor (LDLR) homozygous mutation rats with hypercholesterolemia and wild-type rats.Methods Thirty LDLR-/-rats and 28 wild-type rats were selected.Plasma cholesterol levels were measured by enzyme-linked immunosorbent assay at 15,18 and 26 weeks old respectively.The axonal structure of the corpus callosum area was observed by transmission electron microscopy.The myelin basic protein (MBP) of the corpus callosum area was quantitatively analyzed by Western blotting.In addition,at 26 weeks old,the myelin sheaths were stained by fast blue staining.The expression level of MBP in white matter was further detected by immunofluorescence staining,and the morphological changes of glial cells were observed.Results Compared with the wild-type rats,the plasma cholesterol concentration in LDLR-/-rats increased significantly,and it could be as high as 3.3 times at 26 weeks.The results of electron microscopy showed that the LDLR-/-rats had axonal injury at 15 weeks and aggravated gradually over time.At 26 weeks,Western blot analysis of the LDLR-/-rats showed that the MBP expression level of the corpus callosum area decreased significantly.Fast blue staining showed loosening of nerve fibers,diffuse vacuole formation,and myelinated nerve fiber loss in the corpus callosum area.In addition,it was also found that the number of oligodendrocytes in LDLR-/-rats was significantly reduced,and large numbers of astrocytes and microglia were activated.Conclusions LDLR-/-rats will have spontaneous hypercholesterolemia.Axonal injury,demyelination,decreased oligodendrocytes,as well as the abnormal activation of astrocytes and microglia are present in the early adult brain white matter area.

摘要： 目的 探讨在中国汉族人群中,低密度脂蛋白受体(LDL-R)的基因多态性与非酒精性脂肪性肝病(NAFLD)的相关性,并评估该基因多态性对血脂水平的影响.方法 纳入2015年10月-2017年7月于青岛市市立医院就诊的216例NAFLD患者和135例健康对照组,利用PCR及基因质谱测序检测方法对LDL-R rs1433099和rs2738464位点进行基因型检测.同时利用生物化学方法检测上述受试者血脂水平及肝脏酶谱的变化.通过Hardy-Weinberg(H-W)平衡检验样本的群体代表性,用x2检验分析基因型的分布是否符合H-W平衡.基因型及等位基因频率采用直接计数法;计数资料组间比较采用x2检验或Fisher精确检验分析;计量资料组间比较采用t检验和Wilcoxon秩和检验;rs1433099和rs2738464多态性与NAFLD发生的相对风险度以比值比(0R)及其95％可信区间(95％CI)表示,采用二元logistic回归分析模型计算.结果 LDL-R基因rs1433099和rs2738464位点的等位基因频率及基因型分布频率在NA FLD组和健康对照组中差异均无统计学意义(P值均＞0.05);同时,2组间血脂水平、肝脏酶学、BMI差异均无统计学意义(P值均＞0.05).在NAFLD组、健康对照组及全部纳入者的各组内,rs1433099 T和rs2738464 G的携带者与非携带者之间的一般临床特征,包括血脂水平、BMI、血压情况及肝脏酶谱之间比较,差异均无统计学意义(P值均＞0.05).结论 LDL-R基因的rs1433099和rs2738464位点多态性不影响血脂水平.同时,首次证明了位于LDL-R基因上的rs1433099和rs2738464位点的基因多态性在中国汉族人群中与NAFLD发病风险不相关.

摘要： 目的 探讨无果枸杞芽提取物(fruitless lycium-sprout extracts,FLE)对低密度脂蛋白受体(LDLR)-/-小鼠动脉粥样硬化病程的影响,为开发FLE调控血脂、抑制动脉粥样硬化病程进展的药用价值提供理论依据.方法 选择12周龄雄性LDLR-/-小鼠48只,分为正常饮食组16只,动脉粥样硬化模型组(模型组)16只,FLE组16只.小鼠体质量在研究开始时测量,每4周测量1次,分别于12、16、20和24周时检测,测量体质量当天采集血液,分离血清,检测TC、TG及氧化型低密度脂蛋白(oxLDL)/β2糖蛋白Ⅰ(β2GPI);收集小鼠尿液,试剂盒测定尿液中11-脱氢血栓素B2(11-dhTXB2)含量.24周时用组织化学染色法观察各组小鼠主动脉斑块形成情况,评估FLE的疗效.结果 模型组小鼠16、20、24周时血清TC、TG和尿11-dhTXB2水平明显高于FLE组(P＜0.05,P＜0.01).模型组小鼠20、24周时oxLDL/β2GPI复合物水平明显高于FLE组,差异有统计学意义(P＜0.05,P＜0.01).正常饮食组、模型组和FLE组小鼠24周时动脉斑块面积分别为(3.36±0.87)％、(20.71±2.17)％和(6.16±0.76)％.正常饮食组几乎看不到斑块形成,模型组斑决明显增加,而FLE组斑块面积较模型组明显受到抑制(P＜0.01).结论 FLE能有效抑制高脂饮食诱发的小鼠动脉斑块形成,延缓病程发展.

摘要： Objective To investigate the role of cyclooxygenase-2 (COX-2) in podocyte injury in diabetic rats mediated by the disruption of low-density lipoprotein receptor (LDLr) pathway.Methods Eight-week old male Sprague-Dawley (SD) rats were treated for 12 weeks by dividing into three groups:control rats,streptozotocin (STZ) induced diabetic rats (DM),and diabetic rats treated with aspirin (DM+Aspirin).The plasma lipid profile was checked by clinical biochemistry assay.The ratio of urinary microalbumin to creatinine (ACR) was detected by enzyme-linked immunosorbent assay.Intracellular lipid accumulation was evaluated by Oil Red O staining and a free cholesterol quantitative assay.The glomerular podocyte injury and the expression of molecules related with LDLr pathway were evaluated by electron microscope,immunohistochemical staining,immunofluorescent staining,and Western blotting.Results There were increased levels of urinary ACR (P ＜ 0.01) and podocyte injury(P ＜ 0.01) in DM rats compared with the controls.Additionally,lipid accumulation in kidneys of DM rats were significantly increased (P ＜ 0.01),due to increased protein expressions of COX-2,LDLr,sterol regulatory element-binding protein (SREBP) cleavage activating protein (SCAP),and SREBP-2 (P ＜ 0.01).However,these changes were significantly inhibited by an inhibitor of COX-2,Aspirin (P ＜ 0.05).It's worth noting that,COX-2 protein expression was closely correlated with LDLr protein expression (r=0.85,P ＜ 0.01).Conclusion Dysregulation of LDLr pathway contributes to podocyte injury in diabetic nephropathy,which may be mediated through the increased COX-2 expression.%目的 观察环氧化酶2(cyclooxygenase-2,COX-2)介导的低密度脂蛋白受体(low density lipoprotein receptor,LDLr)表达失调在糖尿病肾病(diabetic nephropathy,DN)早期足细胞损伤中的作用.方法 选取8周龄雄性Sprague-Dawley(SD)大鼠,采用链尿菌素(streptozotocin,STZ)诱导糖尿病大鼠模型,造模成功后按照简单随机抽样原则分为对照(Control)组、糖尿病(diabetic mellitus,DM)组、DM+阿司匹林(Aspirin)组,每组10只.于第12周收集24 h尿液、检测尿微量白蛋白/肌酐比值(microalbumin to creatinine ratio,ACR),并于第12周处死,收集血标本,检测血脂谱,留取肾组织.免疫组化及Western印迹法观察COX-2、LDLr、胆固醇调节元件结合蛋白2(sterol regulatory element-binding protein-2,SREBP-2)及其裂解激活蛋白(SREBP cleavage-activating protein,SCAP)在肾脏的表达情况,免疫荧光法观察COX-2与WT-1在肾脏的共表达情况.透射电镜观察足细胞超微结构的改变,免疫组化及Western印迹法观察足细胞特异性标志蛋白WT-1及nephrin在肾脏的表达情况.分析COX-2与LDLr表达的相关性.油红0染色及胞内游离胆固醇定量测定法观察肾小球脂质沉积情况.结果 与对照组相比,DM组大鼠尿ACR显著升高(P＜0.01),肾脏COX-2及LDLr、SCAP、SREBP-2蛋白的表达显著增加(P＜0.01),足细胞损伤加重(P＜0.01),肾小球内有显著的脂质沉积(P＜ 0.01),而DM+Aspirin组上述改变明显减轻(P＜0.05).并且COX-2的表达与LDLr的表达呈正相关(r=0.85,P＜0.01).激光共聚焦荧光显微镜观察证实,大鼠肾脏COX-2与足细胞特异性标志物WT-1存在共表达.结论 COX-2的过度表达加重了DN足细胞损伤,其机制可能与破坏LDLr负反馈调节、上调肾脏足细胞LDLr表达、增加细胞内胆固醇摄入,致使肾小球足细胞内脂质过度沉积有关.抑制COX-2表达能减轻糖尿病足细胞损伤.

摘要： 人类前蛋白转化酶枯草溶菌素9（PCSK9）是近年来新发现的一种与常染色体显性高胆固醇血症密切相关的基因，编码前蛋白转化酶 NARC-1。PCSK9除了通过调节细胞表面低密度脂蛋白受体（LDLR）水平参与脂质代谢外，亦直接对血管壁细胞发挥毒性作用，从而在动脉粥样硬化的发生、发展中起重要作用。因此，阐明 PCSK9在动脉粥样硬化发生中的作用及机制，对高胆固醇血症和动脉粥样硬化的临床药物应用具有重要意义。

摘要： 目的检测一中国汉族甲状腺功能亢进症(简称甲亢)合并家族性高胆固醇血症(FH)家系低密度脂蛋白受体(LDLR)基因突变情况。方法收集该家系外周血并提取DNA,采用TOUCH-DOWN PCR扩增病人及其家系成员6人LDLR基因启动子和全部18个外显子片段,并进行测序分析,同时检测载脂蛋白B100(apoB100)基因Q3500R、R3531C和R3500W突变情况,以排除家族性apoB100缺陷症。结果该家系共5例病人,符合常染色体显性遗传病的特点,经突变筛查,发现LDLR基因存在6处碱基改变,未导致氨基酸的改变,经Pubmed数据库比对,为单核苷酸多态性位点(rs5925、rs5929、rs5930、rs4508523、rs2738460、rs2116897)。结论 LDLR基因不是该家系的致病基因,需对其他可能的候选致病基因进行筛查。临床医师对甲亢合并血脂紊乱者,应注意遗传因素在脂代谢紊乱中作用,根据基因变异类型制定个体化调脂策略。

摘要： 本发明涉及家蚕型生物素化CTLD14或sRAGE及其制造方法。本发明的一个实施方案提供了用于制造生物素化蛋白质的方法，其中所述方法包括：A）用于将编码生物素连接酶和蛋白质的核酸分子以可共表达的方式插入家蚕或赋予与家蚕的糖链相同的糖链的活生物内的步骤，B）通过将家蚕或赋予与家蚕的糖链相同的糖链的活生物置于核酸分子对于其进行表达的条件下，用于促使生物素连接酶和蛋白质表达的步骤，C）用于将生物素施用于活生物并且获得生物素化蛋白质的步骤。

摘要： 本发明涉及结合人凝集素样氧化的LDL(低密度脂蛋白)受体1(下文中，有时称为“LOX‑1”)的单克隆抗体，以及包含其的药物组合物和治疗方法。

摘要： 本发明涉及家蚕型生物素化CTLD14或sRAGE及其制造方法。本发明的一个实施方案提供了用于制造生物素化蛋白质的方法，其中所述方法包括：A）用于将编码生物素连接酶和蛋白质的核酸分子以可共表达的方式插入家蚕或赋予与家蚕的糖链相同的糖链的活生物内的步骤，B）通过将家蚕或赋予与家蚕的糖链相同的糖链的活生物置于核酸分子对于其进行表达的条件下，用于促使生物素连接酶和蛋白质表达的步骤，C）用于将生物素施用于活生物并且获得生物素化蛋白质的步骤。

摘要： 本发明提供了一种制备基因工程仓鼠的方法，包括将该仓鼠体内的LDL受体基因敲除。在优选实施方案中，所述LDL受体基因通过CRISPR/CAS9基因编辑技术敲除。该基因工程仓鼠表现出明显的血脂升高，可以直接用于高脂血症和动脉硬化研究，是和人类家族性高胆固醇血症高度相似的不可多得的小动物模型。本发明还提供了利用该基因工程仓鼠筛选药物的方法。

摘要： 本发明涉及与人凝集素样氧化型LDL(低密度脂蛋白)受体1(下文，有时称作“LOX-1”)结合的单克隆抗体，和包含所述单克隆抗体的药物组合物和治疗方法。

摘要： 本发明涉及结合人凝集素样氧化的LDL(低密度脂蛋白)受体1(下文中，有时称为“LOX-1”)的单克隆抗体，以及包含其的药物组合物和治疗方法。

摘要： 本发明鉴定了一种具有优异的凝集素样氧化型LDL受体抑制作用的化合物，本发明提供一种凝集素样氧化型LDL抑制用医药品、以及含有三聚体以上的原花青素为有效成分且具有凝集素样氧化型LDL受体抑制作用的动脉硬化预防医药品，所述凝集素样氧化LDL受体抑制用医药品对于治疗或预防那些以凝集素样氧化型LDL受体作为恶化因素发挥作用的疾病、特别是动脉硬化疾病有用，其以三聚体以上的原花青素(Procyanidin)为有效成分，所述原花青素来源于选自苹果、葡萄种子、花生内皮以及松树皮的植物。

摘要： 本发明涉及一种针对人类LDL(低密度脂蛋白)受体的单克隆抗体，其中每条轻链的可变区域是由鼠科核酸序列SEQ ID NO：5编码的，每条重链的可变区域是由鼠科核酸序列SEQ ID NO：7编码的，或者由具有与序列SEQ ID NO：5和SEQ ID NO：7足够同源性的，其本质和抗体与其抗原间的结合亲合力未被改变的序列编码，而其轻链和重链的恒定区域是来自非鼠科种的恒定区域。

摘要： 本发明包括用于靶向免疫细胞上LOX－1受体的组合物和方法，以及抗LOX－1抗体的用途。

摘要： 本发明涉及分析导致家族性高胆固醇血症的突变的存在或缺乏的体外方法。本发明方法描述这样的途径，其中可以使用来自个体的DNA样品检测所述突变，并且该方法包括下列：使用与低密度脂蛋白受体基因互补的引物进行聚合酶链式反应；通过测序分析扩增的产物；限制性酶切分析；单链构象多态性技术；可以用于检测前述DNA中的突变的异源双链分析以及在生物芯片玻璃支持体上的装置的分析，在所述生物芯片玻璃支持体上排列有寡核苷酸探针。