目的:研究蓝靛果花色苷对高脂血症大鼠肝脏低密度脂蛋白受体(low density lipoprotein receptor,LDLR)、三磷酸腺苷结合盒转运体G1(ATP-binding cassette transporter G1,ABCG1)及ABCA1基因表达的影响。方法:选择2月龄雄性Wistar大鼠60...目的:研究蓝靛果花色苷对高脂血症大鼠肝脏低密度脂蛋白受体(low density lipoprotein receptor,LDLR)、三磷酸腺苷结合盒转运体G1(ATP-binding cassette transporter G1,ABCG1)及ABCA1基因表达的影响。方法:选择2月龄雄性Wistar大鼠60只,将大鼠随机分为6组,分别为基础饲料对照组(ND,1.2 g/(kg·dm_b)生理盐水灌胃)、高脂模型对照组(HFD,1.2 g/(kg·dm_b)生理盐水灌胃)、阳性对照组(10 mg/(kg·dm_b)辛伐他汀片灌胃),蓝靛果花色苷低、中、高剂量组(HFD+L、HFD+M、HFD+H,分别给予4.0、40.0、120.0 mg/(kg·dm_b)的花色苷灌胃),持续28 d。实验结束后,测定血清总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)、低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)、载脂蛋白A(apolipoprotein A,Apo-A)及载脂蛋白B(Apo-B)等血脂指标水平。取大鼠肝脏,利用实时荧光定量聚合酶链式反应测定大鼠肝脏组织中LDLR、ABCG1、ABCA1 m RNA表达量,Western blot检测LDLR蛋白表达水平。结果:蓝靛果花色苷干预后,与HFD组相比,花色苷均能不同程度地降低高血脂大鼠血清中TC、TG、LDL-C、Apo-B的含量(P<0.05或P<0.01),显著升高HDL-C及Apo-A的含量(P<0.05或P<0.01)。花色苷各剂量组LDLR蛋白和m RNA水平均增高,与HFD组比较差异有显著性(P<0.05),ABCA1 m RNA和ABCG1 m RNA的表达水平也高于HFD组,尤其是花色苷中、高剂量组差异明显(P<0.05)。结论:40.0 mg/(kg·dm_b)蓝靛果花色苷具有明显的调节血脂作用,其作用机制可能是通过上调肝脏LDLR和ABC家族基因的表达,进而调节胆固醇逆转运过程。展开更多
Single nucleotide polymorphisms （SNP） of ATP-binding cassette transporter A1 （ABCA1） gene are related to plasma lipid and susceptibility to coronary artery disease （CAD）. Our first goal was to screen all 50 codi...Single nucleotide polymorphisms （SNP） of ATP-binding cassette transporter A1 （ABCA1） gene are related to plasma lipid and susceptibility to coronary artery disease （CAD）. Our first goal was to screen all 50 coding regions of ABCA1 to find new SNPs. Our second goal was to investigate the frequency distribution of R1587K and M883I polymorphisms of ABCA1 gene, which are the variant occurred most frequently, in Chinese people and to evaluate their association with the CAD phenotype and plasma lipids. Methods： Single-strand conformation polymorphism （SSCP） and DNA sequence were used for confirming new SNP of ABCA1, and restriction fragment length polymorphism （RFLP） were applied for confirming genotypes of R1587K and M883I in 112 CAD cases and 108 healthy people. Results： We discovered a new ABCA1 SNP in Chinese population, which converse 233 amino acids from Methionine to Valine （M233V）. This new ABCA1 SNP located in exon7, and might potentially modulate the biological function of lipid metabolism. For R1587K and M883I SNPs, the K allele and I allele frequency was 28.9% and 31.1%, respectively. The K allele at R1587K conferred lower mean values of HDL-C in a dose-dependent manner in both CAD patients and healthy people. However, 883I allele was not associated with plasma lipid level. Neither 1587KK nor 883II associated with increased risk of CAD. Conclusion： Our study finds a potential functional ABCA1 SNPs and revealed K allele of R1587K associated decreased HDL-C level in Chinese population.展开更多
Objective To investigate the effects of rapamycin on cholesterol homeostasis of glomerular mesangial cells and the underlying mechanisms. Methods Intracellular cholesterol accumulation was measured by Oil Red O stain...Objective To investigate the effects of rapamycin on cholesterol homeostasis of glomerular mesangial cells and the underlying mechanisms. Methods Intracellular cholesterol accumulation was measured by Oil Red O staining and high performance liquid chromatography. The effects of rapamycin on interleukin-1β（1L-1β）-induced mRNA and protein changes of low-density lipoprotein receptor （LDLR） and ATP-binding cassette transporter Al （ABCAl） were assayed by quantitative real-time PCR and Western blot. Transient expressions of 3 types of mammalian target of rapamycin （mTOR）, including mTOR-WT （wild type）, mTOR-RR （rapamycin resistant, with kinase activity）, and mTOR-RR-KD （rapamycin resistant, without kinase activity）, were obtained by plasmid transfection. Results Rapamycin had no significant influence on intracellular cholesterol concentration trader normal condition, but it significantly decreased the intracellular cholesterol concentration in the presence of IL-1β. Rapamycin dose-dependently suppressed the increased expression of LDLR induced by IL-1β and up-regulated the suppressed expression of ABCAl caused by IL-1β Transient expression of 3 types of mTOR all reduced ABCAl mRNA expression significantly, which all could be overroded by rapamycin. Conclusions Rapamycin may contribute to the maintaining of glomerular mesangial cell intracellular cholesterol homeostasis under inflammatory state by both reducing cholesterol uptake and increasing cholesterol effiux. And the effect may be not completely mediiated by mTOR.展开更多
Objective The current study aimed to evaluate whether the induction of macrophage inflammatory cytokines by Ox-LDL is related to the expression of ABCA 1 pathway. Methods After THP 1/PMA macrophages were transfected w...Objective The current study aimed to evaluate whether the induction of macrophage inflammatory cytokines by Ox-LDL is related to the expression of ABCA 1 pathway. Methods After THP 1/PMA macrophages were transfected with ABCA1 antisense oligonucleotides （100nmol/L） followed by treatment with Ox-LDL （30mg/L）, the expressions of ABCA1, ICAM-1 and MCP-1 mRNA and protein were determined by real-time fluorescent quantitative RT-PCR, Western blot or ELISA. Results Ox-LDL induced expressions of ABCA1, ICAM-1, and MCP-1 at both mRNA and protein levels from THPI/PMA macrophages. Transfection with ABCAI antisense oligonucleotides reduced ABCA1 mRNA levels after 3 and 6 hours and protein levels after 12 and 24 hours. The expression of ICAM-1 and MCP-1 induced by Ox-LDL was also decreased after inhibition of ABCA 1 protein expression by ABCA 1 antisense oligonucleotide decreased. Conclusion The induction of macrophage inflammatory cytokines by Ox-LDL is partially dependent on expression ofABCA1. Our studies disclose new functions of ABCA1 in macrophages Objective The current study aimed to evaluate whether the induction of macrophage inflammatory eytokines by Ox-LDL is related to the expression of ABCA 1 pathway. Methods After THP 1/PMA macrophages were transfected with ABCA1 antisense oligonucleotides （100nmol/L） followed by treatment with Ox-LDL （30mg/L）, the expressions of ABCA1, ICAM-1 and MCP-1 mRNA and protein were determined by real-time fluorescent quantitative RT-PCR, Western blot or ELISA. Results Ox-LDL induced expressions of ABCA1, ICAM-1, and MCP-1 at both mRNA and protein levels from THPI/PMA macrophages. Transfection with ABCAI antisense oligonucleotides reduced ABCA1 mRNA levels after 3 and 6 hours and protein levels after 12 and 24 hours. The expression of ICAM-1 and MCP-1 induced by Ox-LDL was also decreased after inhibition of ABCA 1 protein expression by ABCA 1 antisense oligonucleotide decreased. Conclusion The induction of macrophage inflammatory cytokines by Ox-LDL is partially dependent on expression ofABCA1. Our studies disclose new functions of ABCA1 in macrophages展开更多
目的探讨人参炔三醇(panaxytriol,PXT)抑制氧化低密度脂蛋白(oxidized low density lipoprotein,oxLDL)诱导单核巨噬细胞脂质沉积的作用机制。方法单核细胞THP-1经佛波酯(phorbol-12-myristate-13-acetate,PMA)诱导成巨噬细胞后,采用不...目的探讨人参炔三醇(panaxytriol,PXT)抑制氧化低密度脂蛋白(oxidized low density lipoprotein,oxLDL)诱导单核巨噬细胞脂质沉积的作用机制。方法单核细胞THP-1经佛波酯(phorbol-12-myristate-13-acetate,PMA)诱导成巨噬细胞后,采用不同浓度(0、1、5、10、20μmol/L)PXT处理单核巨噬细胞24小时,通过MTT法检测其对细胞存活率的影响。THP-1单核巨噬细胞经PMA刺激后,利用oxLDL诱导脂质蓄积,PXT处理后利用流式细胞仪,荧光显微镜和油红O染色检测巨噬细胞对oxLDL的摄取。采用Western blot检测细胞内白细胞分化抗原36(cluster of differentiation,CD36)、三磷酸腺苷结合盒转运体A1(ATP-binding cassette transporter A1,ABCA1)、清道夫受体B1(scavenger receptor class B type 1,SRB1)的蛋白表达水平变化。结果MTT实验结果显示,与正常组比较,20μmol/L PXT处理THP-1单核巨噬细胞24小时后显著抑制了细胞活力(P<0.05),且其他浓度对细胞活力均无显著影响。流式细胞检测、细胞荧光成像和油红O染色结果显示,与模型组比较,PXT浓度依赖地降低巨噬细胞内DiI-oxLDL荧光强度与细胞内脂质蓄积(P<0.05)。Western blot结果显示,与模型组比较,PXT浓度依赖地降低了巨噬细胞中CD36蛋白的表达(P<0.05),而对其他蛋白无显著影响。结论PXT通过抑制清道夫受体CD36的表达,抑制巨噬细胞对oxLDL的吞噬及泡沫细胞的形成,具有潜在的抗动脉粥样硬化活性。展开更多
文摘目的:研究蓝靛果花色苷对高脂血症大鼠肝脏低密度脂蛋白受体(low density lipoprotein receptor,LDLR)、三磷酸腺苷结合盒转运体G1(ATP-binding cassette transporter G1,ABCG1)及ABCA1基因表达的影响。方法:选择2月龄雄性Wistar大鼠60只,将大鼠随机分为6组,分别为基础饲料对照组(ND,1.2 g/(kg·dm_b)生理盐水灌胃)、高脂模型对照组(HFD,1.2 g/(kg·dm_b)生理盐水灌胃)、阳性对照组(10 mg/(kg·dm_b)辛伐他汀片灌胃),蓝靛果花色苷低、中、高剂量组(HFD+L、HFD+M、HFD+H,分别给予4.0、40.0、120.0 mg/(kg·dm_b)的花色苷灌胃),持续28 d。实验结束后,测定血清总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)、低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)、载脂蛋白A(apolipoprotein A,Apo-A)及载脂蛋白B(Apo-B)等血脂指标水平。取大鼠肝脏,利用实时荧光定量聚合酶链式反应测定大鼠肝脏组织中LDLR、ABCG1、ABCA1 m RNA表达量,Western blot检测LDLR蛋白表达水平。结果:蓝靛果花色苷干预后,与HFD组相比,花色苷均能不同程度地降低高血脂大鼠血清中TC、TG、LDL-C、Apo-B的含量(P<0.05或P<0.01),显著升高HDL-C及Apo-A的含量(P<0.05或P<0.01)。花色苷各剂量组LDLR蛋白和m RNA水平均增高,与HFD组比较差异有显著性(P<0.05),ABCA1 m RNA和ABCG1 m RNA的表达水平也高于HFD组,尤其是花色苷中、高剂量组差异明显(P<0.05)。结论:40.0 mg/(kg·dm_b)蓝靛果花色苷具有明显的调节血脂作用,其作用机制可能是通过上调肝脏LDLR和ABC家族基因的表达,进而调节胆固醇逆转运过程。
基金Supported by the Natural Scientific Foundation of China (30471929)
文摘Single nucleotide polymorphisms （SNP） of ATP-binding cassette transporter A1 （ABCA1） gene are related to plasma lipid and susceptibility to coronary artery disease （CAD）. Our first goal was to screen all 50 coding regions of ABCA1 to find new SNPs. Our second goal was to investigate the frequency distribution of R1587K and M883I polymorphisms of ABCA1 gene, which are the variant occurred most frequently, in Chinese people and to evaluate their association with the CAD phenotype and plasma lipids. Methods： Single-strand conformation polymorphism （SSCP） and DNA sequence were used for confirming new SNP of ABCA1, and restriction fragment length polymorphism （RFLP） were applied for confirming genotypes of R1587K and M883I in 112 CAD cases and 108 healthy people. Results： We discovered a new ABCA1 SNP in Chinese population, which converse 233 amino acids from Methionine to Valine （M233V）. This new ABCA1 SNP located in exon7, and might potentially modulate the biological function of lipid metabolism. For R1587K and M883I SNPs, the K allele and I allele frequency was 28.9% and 31.1%, respectively. The K allele at R1587K conferred lower mean values of HDL-C in a dose-dependent manner in both CAD patients and healthy people. However, 883I allele was not associated with plasma lipid level. Neither 1587KK nor 883II associated with increased risk of CAD. Conclusion： Our study finds a potential functional ABCA1 SNPs and revealed K allele of R1587K associated decreased HDL-C level in Chinese population.
文摘Objective To investigate the effects of rapamycin on cholesterol homeostasis of glomerular mesangial cells and the underlying mechanisms. Methods Intracellular cholesterol accumulation was measured by Oil Red O staining and high performance liquid chromatography. The effects of rapamycin on interleukin-1β（1L-1β）-induced mRNA and protein changes of low-density lipoprotein receptor （LDLR） and ATP-binding cassette transporter Al （ABCAl） were assayed by quantitative real-time PCR and Western blot. Transient expressions of 3 types of mammalian target of rapamycin （mTOR）, including mTOR-WT （wild type）, mTOR-RR （rapamycin resistant, with kinase activity）, and mTOR-RR-KD （rapamycin resistant, without kinase activity）, were obtained by plasmid transfection. Results Rapamycin had no significant influence on intracellular cholesterol concentration trader normal condition, but it significantly decreased the intracellular cholesterol concentration in the presence of IL-1β. Rapamycin dose-dependently suppressed the increased expression of LDLR induced by IL-1β and up-regulated the suppressed expression of ABCAl caused by IL-1β Transient expression of 3 types of mTOR all reduced ABCAl mRNA expression significantly, which all could be overroded by rapamycin. Conclusions Rapamycin may contribute to the maintaining of glomerular mesangial cell intracellular cholesterol homeostasis under inflammatory state by both reducing cholesterol uptake and increasing cholesterol effiux. And the effect may be not completely mediiated by mTOR.
文摘Objective The current study aimed to evaluate whether the induction of macrophage inflammatory cytokines by Ox-LDL is related to the expression of ABCA 1 pathway. Methods After THP 1/PMA macrophages were transfected with ABCA1 antisense oligonucleotides （100nmol/L） followed by treatment with Ox-LDL （30mg/L）, the expressions of ABCA1, ICAM-1 and MCP-1 mRNA and protein were determined by real-time fluorescent quantitative RT-PCR, Western blot or ELISA. Results Ox-LDL induced expressions of ABCA1, ICAM-1, and MCP-1 at both mRNA and protein levels from THPI/PMA macrophages. Transfection with ABCAI antisense oligonucleotides reduced ABCA1 mRNA levels after 3 and 6 hours and protein levels after 12 and 24 hours. The expression of ICAM-1 and MCP-1 induced by Ox-LDL was also decreased after inhibition of ABCA 1 protein expression by ABCA 1 antisense oligonucleotide decreased. Conclusion The induction of macrophage inflammatory cytokines by Ox-LDL is partially dependent on expression ofABCA1. Our studies disclose new functions of ABCA1 in macrophages Objective The current study aimed to evaluate whether the induction of macrophage inflammatory eytokines by Ox-LDL is related to the expression of ABCA 1 pathway. Methods After THP 1/PMA macrophages were transfected with ABCA1 antisense oligonucleotides （100nmol/L） followed by treatment with Ox-LDL （30mg/L）, the expressions of ABCA1, ICAM-1 and MCP-1 mRNA and protein were determined by real-time fluorescent quantitative RT-PCR, Western blot or ELISA. Results Ox-LDL induced expressions of ABCA1, ICAM-1, and MCP-1 at both mRNA and protein levels from THPI/PMA macrophages. Transfection with ABCAI antisense oligonucleotides reduced ABCA1 mRNA levels after 3 and 6 hours and protein levels after 12 and 24 hours. The expression of ICAM-1 and MCP-1 induced by Ox-LDL was also decreased after inhibition of ABCA 1 protein expression by ABCA 1 antisense oligonucleotide decreased. Conclusion The induction of macrophage inflammatory cytokines by Ox-LDL is partially dependent on expression ofABCA1. Our studies disclose new functions of ABCA1 in macrophages
文摘目的探讨人参炔三醇(panaxytriol,PXT)抑制氧化低密度脂蛋白(oxidized low density lipoprotein,oxLDL)诱导单核巨噬细胞脂质沉积的作用机制。方法单核细胞THP-1经佛波酯(phorbol-12-myristate-13-acetate,PMA)诱导成巨噬细胞后,采用不同浓度(0、1、5、10、20μmol/L)PXT处理单核巨噬细胞24小时,通过MTT法检测其对细胞存活率的影响。THP-1单核巨噬细胞经PMA刺激后,利用oxLDL诱导脂质蓄积,PXT处理后利用流式细胞仪,荧光显微镜和油红O染色检测巨噬细胞对oxLDL的摄取。采用Western blot检测细胞内白细胞分化抗原36(cluster of differentiation,CD36)、三磷酸腺苷结合盒转运体A1(ATP-binding cassette transporter A1,ABCA1)、清道夫受体B1(scavenger receptor class B type 1,SRB1)的蛋白表达水平变化。结果MTT实验结果显示,与正常组比较,20μmol/L PXT处理THP-1单核巨噬细胞24小时后显著抑制了细胞活力(P<0.05),且其他浓度对细胞活力均无显著影响。流式细胞检测、细胞荧光成像和油红O染色结果显示,与模型组比较,PXT浓度依赖地降低巨噬细胞内DiI-oxLDL荧光强度与细胞内脂质蓄积(P<0.05)。Western blot结果显示,与模型组比较,PXT浓度依赖地降低了巨噬细胞中CD36蛋白的表达(P<0.05),而对其他蛋白无显著影响。结论PXT通过抑制清道夫受体CD36的表达,抑制巨噬细胞对oxLDL的吞噬及泡沫细胞的形成,具有潜在的抗动脉粥样硬化活性。
