Creutzfeldt-Jakob disease(CJD)is a rare neurodegenerative disorder characterized by abnormalities in the prion protein(PrP),the most common form of human prion disease.Although Genome-Wide Association Studies(GWAS)hav...Creutzfeldt-Jakob disease(CJD)is a rare neurodegenerative disorder characterized by abnormalities in the prion protein(PrP),the most common form of human prion disease.Although Genome-Wide Association Studies(GWAS)have identified numerous risk genes for CJD,the mechanisms underlying these risk loci remain poorly understood.This study aims to elucidate novel genetically prioritized candidate proteins associated with CJD in the human brain through an integrative analytical pipeline.Utilizing datasets from Protein Quantitative Trait Loci(pQTL)(NpQTL1=152,NpQTL2=376),expression QTL(eQTL)(N=452),and the CJD GWAS(NCJD=4110,NControls=13569),we implemented a systematic analytical pipeline.This pipeline included Proteome-Wide Association Study(PWAS),Mendelian randomization(MR),Bayesian colocalization,and Transcriptome-Wide Association Study(TWAS)to identify novel genetically prioritized candidate proteins implicated in CJD pathogenesis within the brain.Through PWAS,we identified that the altered abundance of six brain proteins was significantly associated with CJD.Two genes,STX6 and PDIA4,were established as lead causal genes for CJD,supported by robust evidence(False Discovery Rate<0.05 in MR analysis;PP4/(PP3+PP4)≥0.75 in Bayesian colocalization).Specifically,elevated levels of STX6 and PDIA4 were associated with an increased risk of CJD.Additionally,TWAS demonstrated that STX6 and PDIA4 were associated with CJD at the transcriptional level.展开更多
目的构建糖尿病肾病大鼠模型,从整体水平观察川陈皮素对其的治疗作用,并从分子水平研究川陈皮素对丝氨酸蛋白酶家族B成员7(serpin family B member 7,Megsin)、血小板衍生生长因子-BB(platelet-derived growth factor-BB,PDGF-BB)、细...目的构建糖尿病肾病大鼠模型,从整体水平观察川陈皮素对其的治疗作用,并从分子水平研究川陈皮素对丝氨酸蛋白酶家族B成员7(serpin family B member 7,Megsin)、血小板衍生生长因子-BB(platelet-derived growth factor-BB,PDGF-BB)、细胞外信号调节蛋白激酶1/2(extracellular signal-regulated protein kinase,ERK1/2)和Ⅳ型胶原(collagen IV)蛋白及mRNA表达的影响。方法清洁级雄性SD大鼠120只分为正常组、糖尿病肾病模型组、贝那普利组、川陈皮素低、中和高剂量组,高脂高糖饮食1月+一次性腹腔注射链尿佐菌素(streptozotocin,STZ)诱导糖尿病肾病大鼠模型,相应药物治疗6周后处死,收集尿液、血液和肾脏,检测血糖、血清和尿液中肌酐和β2-MG、小鼠肾重与体重比(KW/BW),同时检测肾脏炎性因子IL-1、IL-6和TNF-α含量,收集肾组织标本,聚合酶链式反应和蛋白印记法测定肾组织中Megsin、PDGF-BB、pERK1/2及collagen IV的表达。结果肾脏组织HE染色可见,正常组大鼠形态正常,模型组出现明显肾小球萎缩和硬化,与模型组相比,贝那普利组、川陈皮素低、中和高剂量组均明显好转,此外贝那普利组和川陈皮素高剂量组基本一致;与正常组相比,模型组大鼠血糖、KW/BW、尿液肾功能指标UREA和β2-MG、血液肾功能指标UREA和β2-MG、肾脏炎性因子IL-1、IL-6和TNF-α、肾脏组织中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA均明显升高(P<0.05),与模型组相比,贝那普利组、川陈皮素低、中和高剂量组血糖、KW/BW、尿液肾功能指标UREA和β2-MG、血液肾功能指标UREA和β2-MG、肾脏炎性因子IL-1、IL-6和TNF-α、肾脏组织中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA均降低(P<0.05),此外川陈皮素各组血糖均明显低于贝那普利组(P<0.05),川陈皮素高剂量组IL-1、IL-6和TNF-α低于贝那普利组(P<0.05),但KW/BW、尿液UREA和β2-MG、血液UREA和β2-MG、肾脏中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA和贝那普利组相比没有差异。结论川陈皮素对糖尿病肾病大鼠有明显的治疗作用,这与川陈皮素可调节肾脏中megsin、PDGF-BB、pERK1/2及collagen IV的表达有关。展开更多
文摘Creutzfeldt-Jakob disease(CJD)is a rare neurodegenerative disorder characterized by abnormalities in the prion protein(PrP),the most common form of human prion disease.Although Genome-Wide Association Studies(GWAS)have identified numerous risk genes for CJD,the mechanisms underlying these risk loci remain poorly understood.This study aims to elucidate novel genetically prioritized candidate proteins associated with CJD in the human brain through an integrative analytical pipeline.Utilizing datasets from Protein Quantitative Trait Loci(pQTL)(NpQTL1=152,NpQTL2=376),expression QTL(eQTL)(N=452),and the CJD GWAS(NCJD=4110,NControls=13569),we implemented a systematic analytical pipeline.This pipeline included Proteome-Wide Association Study(PWAS),Mendelian randomization(MR),Bayesian colocalization,and Transcriptome-Wide Association Study(TWAS)to identify novel genetically prioritized candidate proteins implicated in CJD pathogenesis within the brain.Through PWAS,we identified that the altered abundance of six brain proteins was significantly associated with CJD.Two genes,STX6 and PDIA4,were established as lead causal genes for CJD,supported by robust evidence(False Discovery Rate<0.05 in MR analysis;PP4/(PP3+PP4)≥0.75 in Bayesian colocalization).Specifically,elevated levels of STX6 and PDIA4 were associated with an increased risk of CJD.Additionally,TWAS demonstrated that STX6 and PDIA4 were associated with CJD at the transcriptional level.
文摘泛素连接酶E4B通过U-box基序可将经泛素活化酶(Ubiquitin-activating enzyme, E1)和泛素结合酶(Ubiquitin-conjugating enzyme, E2)传递的泛素(Ubiquitin, UB)标记至底物蛋白质。探究3个蛋白质:金属蛋白酶M20家族蛋白质2(Peptidase M20 domain-containing protein 2,PM20D2)、多腺苷酸结合蛋白质1(Polyadenylate-binding protein 1,PABPC1)、细胞色素C氧化酶Ⅰ翻译激活剂(Translational activator of cytochrome C oxidase Ⅰ,TACO1)是否可被E4B介导泛素化。从HEK293细胞中提取总RNA,反转录为cDNA,以其为模板调取底物基因并构建重组质粒,利用大肠杆菌表达系统表达并纯化泛素化过程所需的各个相关蛋白质,通过蛋白质体外泛素化实验,对3个蛋白质进行泛素化验证。结果表明3个蛋白质均可以在蛋白质水平被E4B泛素化,证明3个蛋白质都是E4B的底物,为进一步在细胞中研究其泛素化的机理奠定基础。
文摘目的构建糖尿病肾病大鼠模型,从整体水平观察川陈皮素对其的治疗作用,并从分子水平研究川陈皮素对丝氨酸蛋白酶家族B成员7(serpin family B member 7,Megsin)、血小板衍生生长因子-BB(platelet-derived growth factor-BB,PDGF-BB)、细胞外信号调节蛋白激酶1/2(extracellular signal-regulated protein kinase,ERK1/2)和Ⅳ型胶原(collagen IV)蛋白及mRNA表达的影响。方法清洁级雄性SD大鼠120只分为正常组、糖尿病肾病模型组、贝那普利组、川陈皮素低、中和高剂量组,高脂高糖饮食1月+一次性腹腔注射链尿佐菌素(streptozotocin,STZ)诱导糖尿病肾病大鼠模型,相应药物治疗6周后处死,收集尿液、血液和肾脏,检测血糖、血清和尿液中肌酐和β2-MG、小鼠肾重与体重比(KW/BW),同时检测肾脏炎性因子IL-1、IL-6和TNF-α含量,收集肾组织标本,聚合酶链式反应和蛋白印记法测定肾组织中Megsin、PDGF-BB、pERK1/2及collagen IV的表达。结果肾脏组织HE染色可见,正常组大鼠形态正常,模型组出现明显肾小球萎缩和硬化,与模型组相比,贝那普利组、川陈皮素低、中和高剂量组均明显好转,此外贝那普利组和川陈皮素高剂量组基本一致;与正常组相比,模型组大鼠血糖、KW/BW、尿液肾功能指标UREA和β2-MG、血液肾功能指标UREA和β2-MG、肾脏炎性因子IL-1、IL-6和TNF-α、肾脏组织中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA均明显升高(P<0.05),与模型组相比,贝那普利组、川陈皮素低、中和高剂量组血糖、KW/BW、尿液肾功能指标UREA和β2-MG、血液肾功能指标UREA和β2-MG、肾脏炎性因子IL-1、IL-6和TNF-α、肾脏组织中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA均降低(P<0.05),此外川陈皮素各组血糖均明显低于贝那普利组(P<0.05),川陈皮素高剂量组IL-1、IL-6和TNF-α低于贝那普利组(P<0.05),但KW/BW、尿液UREA和β2-MG、血液UREA和β2-MG、肾脏中Megsin、PDGF-BB、pERK1/2和collagen IV蛋白与mRNA和贝那普利组相比没有差异。结论川陈皮素对糖尿病肾病大鼠有明显的治疗作用,这与川陈皮素可调节肾脏中megsin、PDGF-BB、pERK1/2及collagen IV的表达有关。
