鼠源抗人乳腺癌多药耐药相关蛋白1(multidrug resistance-associated protein 1,MRP1/ABCC1)噬菌体Fab抗体库的构建首先是利用乳腺癌组织匀浆液作为免疫原免疫BALB/c小鼠,成功免疫后提取小鼠脾组织的总RNA逆转录为cDNA。然后设计小鼠Ig...鼠源抗人乳腺癌多药耐药相关蛋白1(multidrug resistance-associated protein 1,MRP1/ABCC1)噬菌体Fab抗体库的构建首先是利用乳腺癌组织匀浆液作为免疫原免疫BALB/c小鼠,成功免疫后提取小鼠脾组织的总RNA逆转录为cDNA。然后设计小鼠IgG各亚型的特异性Fab引物,PCR扩增Fd和L基因并依次连接到噬菌体载体pComb3中。重组体pComb3-Fab转化至E.coli XL1-Blue菌中,最后经辅助噬菌体VCSM13超感染,成功构建噬菌体Fab抗体库。以化学合成的MRP1/ABCC1膜表位10肽为抗原进行3轮淘选、富集,对淘选后获得的各级噬菌体抗体库进行ELISA检测和鉴定。成功获得的抗MRP1/ABCC1-10肽三级噬菌体Fab抗体库将为抗人乳腺癌MRP1/ABCC1 Fab抗体的制备提供保障。展开更多
目的探讨白三烯受体拮抗剂(leukotriene receptor antagonists,LTRA)与ATP结合盒亚家族C1(ATP-binding cassette sub-family C member 1,ABCC1)基因rs119774位点、溶质载体有机阴离子转运蛋白家族成员2B1(solute carrier organic anion ...目的探讨白三烯受体拮抗剂(leukotriene receptor antagonists,LTRA)与ATP结合盒亚家族C1(ATP-binding cassette sub-family C member 1,ABCC1)基因rs119774位点、溶质载体有机阴离子转运蛋白家族成员2B1(solute carrier organic anion transporter family member 2B1,SLCO2B1)基因rs12422149位点的单核苷酸多态性(single nucleotide polymorphism,SNP)治疗汉族儿童支气管哮喘疗效的相关性。方法选取2019年4月至2021年12月于福建中医药大学附属厦门市中医院就诊的4~13岁汉族支气管哮喘患儿100例为研究对象,采用一代测序法对患儿的ABCC1基因rs119774位点、SLCO2B1基因rs12422149位点进行检测,分析比较2个位点的SNP分布情况。入组患儿均应用LTRA进行治疗,分别于治疗前与治疗12周时进行肺通气功能及脉冲震荡功能检测,并评估临床症状控制水平,分析患儿疗效与SNP的相关性。统计学方法采用t检验、χ^(2)检验。结果治疗前与治疗12周后,ABCC1基因rs119774位点CC型哮喘患儿应用孟鲁司特钠第1秒用力呼气容积(forced expiratory volume in the first second,FEV_(1))[(87.0±16.5)与(93.2±15.1)L,t=-2.071,P=0.038]、峰值呼气流速(peak expiratory flow,PEF)[(77.1±16.9)与(86.5±16.6)L/s,t=-3.233,P=0.001]和25%肺活量的呼气流速(forced expiratory flow 25,FEF25)[(69.6±18.8)与(78.1±19.9)L/s,t=-1.985,P=0.047]均有明显提高,脉冲震荡各项指标显著改善(P<0.05)。治疗后SLCO2B1基因rs12422149位点AA型肺功能指标PEF高于治疗前[(79.0±16.0)与(93.0±9.4)L/s,t=-2.547,P=0.011],治疗后GG型肺功能各指标显著高于治疗前(P<0.05)。ABCC1治疗前后控制率[59%(59/100)与78%(78/100),78%(78/100),χ^(2)=20.964,P<0.001];治疗前后SLCO2B1基因rs119774位点AA型、AG型、GG型的控制率分别为[59%(59/100)与78%(78/100),78%(78/100),χ^(2)=20.964,P<0.001]、[53%(8/15)与80%(12/15),93%(14/15)χ^(2)=12.786,P=0.002]与[56%(23/41)与78%(32/41),78%(32/41),χ^(2)=6.583,P=0.037]与[73%(32/44)与77%(34/44),73%(32/44),χ^(2)=6.471,P=0.039],均显著改善。结论LTRA治疗汉族支气管哮喘患儿与ABCC1、SLCO2B1基因SNP的效果存在相关性。展开更多
Atherosclerosis is a chronic vascular disease closely associated with endothelial dysfunction.Ferroptosis,a major factor in endothelial dysfunction,plays a pivotal role in the progression of atherosclerosis.The develo...Atherosclerosis is a chronic vascular disease closely associated with endothelial dysfunction.Ferroptosis,a major factor in endothelial dysfunction,plays a pivotal role in the progression of atherosclerosis.The development of drugs targeting endothelial ferroptosis offers a potential therapeutic approach for atherosclerosis.This study aimed to assess the potential impact of tetrahydroberberrubine(THBru)on atherosclerosis and unravel its molecular mechanism underlying endothelial protection.Our results demonstrated that THBru significantly reduced plaque formation in the aortas of atherosclerotic mice.Through transcriptome sequencing and further verification,we observed that THBru mitigated endothelial ferroptosis in atherosclerosis by enhancing glutathione homeostasis and decreasing reactive oxygen species(ROS)accumulation.Mechanistically,bioinformatic analysis demonstrated that THBru reduced the expression of the super-enhancer(SE)regulatory gene ATP-binding cassette subfamily C member 1(ABCC1).The transcription factor BTB and CNC homology 1(BACH1)was responsible for ABCC1 transcription by binding to its SE(ABCC1-SE),whereas THBru effectively inhibited the activity of ABCC1-SE.Furthermore,THBru promoted adenosine monophosphate-activated protein kinase(AMPK)activation,thereby negatively regulating BACH1 and the downstream ABCC1/ferroptosis signaling pathway.Collectively,these findings highlight THBru as a promising candidate for treating atherosclerosis,featuring a novel mechanism that inhibits endothelial ferroptosis through the AMPK/BACH1 axis to regulate ABCC1-SE.展开更多
文摘目的探讨白三烯受体拮抗剂(leukotriene receptor antagonists,LTRA)与ATP结合盒亚家族C1(ATP-binding cassette sub-family C member 1,ABCC1)基因rs119774位点、溶质载体有机阴离子转运蛋白家族成员2B1(solute carrier organic anion transporter family member 2B1,SLCO2B1)基因rs12422149位点的单核苷酸多态性(single nucleotide polymorphism,SNP)治疗汉族儿童支气管哮喘疗效的相关性。方法选取2019年4月至2021年12月于福建中医药大学附属厦门市中医院就诊的4~13岁汉族支气管哮喘患儿100例为研究对象,采用一代测序法对患儿的ABCC1基因rs119774位点、SLCO2B1基因rs12422149位点进行检测,分析比较2个位点的SNP分布情况。入组患儿均应用LTRA进行治疗,分别于治疗前与治疗12周时进行肺通气功能及脉冲震荡功能检测,并评估临床症状控制水平,分析患儿疗效与SNP的相关性。统计学方法采用t检验、χ^(2)检验。结果治疗前与治疗12周后,ABCC1基因rs119774位点CC型哮喘患儿应用孟鲁司特钠第1秒用力呼气容积(forced expiratory volume in the first second,FEV_(1))[(87.0±16.5)与(93.2±15.1)L,t=-2.071,P=0.038]、峰值呼气流速(peak expiratory flow,PEF)[(77.1±16.9)与(86.5±16.6)L/s,t=-3.233,P=0.001]和25%肺活量的呼气流速(forced expiratory flow 25,FEF25)[(69.6±18.8)与(78.1±19.9)L/s,t=-1.985,P=0.047]均有明显提高,脉冲震荡各项指标显著改善(P<0.05)。治疗后SLCO2B1基因rs12422149位点AA型肺功能指标PEF高于治疗前[(79.0±16.0)与(93.0±9.4)L/s,t=-2.547,P=0.011],治疗后GG型肺功能各指标显著高于治疗前(P<0.05)。ABCC1治疗前后控制率[59%(59/100)与78%(78/100),78%(78/100),χ^(2)=20.964,P<0.001];治疗前后SLCO2B1基因rs119774位点AA型、AG型、GG型的控制率分别为[59%(59/100)与78%(78/100),78%(78/100),χ^(2)=20.964,P<0.001]、[53%(8/15)与80%(12/15),93%(14/15)χ^(2)=12.786,P=0.002]与[56%(23/41)与78%(32/41),78%(32/41),χ^(2)=6.583,P=0.037]与[73%(32/44)与77%(34/44),73%(32/44),χ^(2)=6.471,P=0.039],均显著改善。结论LTRA治疗汉族支气管哮喘患儿与ABCC1、SLCO2B1基因SNP的效果存在相关性。
基金supported by a grant from NIH(R01 CA120221)China Scholarship Council and Scholarship Award for Excellent Doctoral Student granted by Ministry of Education of China
基金supported by the Noncommunicable Chronic Diseases National Science and Technology Major Project(2024ZD0537908,China)the National Natural Science Foundation of China(82273919,82270396,82570453,82500553,U24A20813 and U21A20339)the Science Foundation for the Excellent Youth Scholars of Heilongjiang Province(JJ2023YX0509,China).
文摘Atherosclerosis is a chronic vascular disease closely associated with endothelial dysfunction.Ferroptosis,a major factor in endothelial dysfunction,plays a pivotal role in the progression of atherosclerosis.The development of drugs targeting endothelial ferroptosis offers a potential therapeutic approach for atherosclerosis.This study aimed to assess the potential impact of tetrahydroberberrubine(THBru)on atherosclerosis and unravel its molecular mechanism underlying endothelial protection.Our results demonstrated that THBru significantly reduced plaque formation in the aortas of atherosclerotic mice.Through transcriptome sequencing and further verification,we observed that THBru mitigated endothelial ferroptosis in atherosclerosis by enhancing glutathione homeostasis and decreasing reactive oxygen species(ROS)accumulation.Mechanistically,bioinformatic analysis demonstrated that THBru reduced the expression of the super-enhancer(SE)regulatory gene ATP-binding cassette subfamily C member 1(ABCC1).The transcription factor BTB and CNC homology 1(BACH1)was responsible for ABCC1 transcription by binding to its SE(ABCC1-SE),whereas THBru effectively inhibited the activity of ABCC1-SE.Furthermore,THBru promoted adenosine monophosphate-activated protein kinase(AMPK)activation,thereby negatively regulating BACH1 and the downstream ABCC1/ferroptosis signaling pathway.Collectively,these findings highlight THBru as a promising candidate for treating atherosclerosis,featuring a novel mechanism that inhibits endothelial ferroptosis through the AMPK/BACH1 axis to regulate ABCC1-SE.
