Niemann-Pick C1 Like 1(NPC1L1)是肠道胆固醇吸收的关键蛋白,也是胆固醇吸收抑制剂依泽替米贝的分子作用靶点。NPC1L1的表达受一些核因子受体调控。敲除apoE-/-小鼠NPC1L1基因能显著抑制动脉粥样硬化的发生和发展,为动脉粥样硬化和冠...Niemann-Pick C1 Like 1(NPC1L1)是肠道胆固醇吸收的关键蛋白,也是胆固醇吸收抑制剂依泽替米贝的分子作用靶点。NPC1L1的表达受一些核因子受体调控。敲除apoE-/-小鼠NPC1L1基因能显著抑制动脉粥样硬化的发生和发展,为动脉粥样硬化和冠状动脉性心脏病提供了新的治疗靶点。展开更多
目的:研究尼曼匹克C1样1(Niemann Pick C1 like 1,NPC1L1)基因启动子-762T>C多态性的活性差异及药物对其的调节作用。方法:分别构建含NPC1L1基因-762T>C多态性的T/C等位基因启动子荧光酶报告基因,测定启动子活性以及不同药物处理...目的:研究尼曼匹克C1样1(Niemann Pick C1 like 1,NPC1L1)基因启动子-762T>C多态性的活性差异及药物对其的调节作用。方法:分别构建含NPC1L1基因-762T>C多态性的T/C等位基因启动子荧光酶报告基因,测定启动子活性以及不同药物处理后启动子活性的改变。结果:含T或C等位基因的NPC1L1基因启动子活性无统计学差异(P>0.05)。外源性胆固醇和依泽麦布均在高浓度抑制两种启动子活性,辛伐他汀则增强两种启动子活性(P<0.05),但两种基因型启动子之间对药物依泽麦布和辛伐他汀的反应无统计学差异(P>0.05)。结论:胆固醇和依泽麦布能抑制NPC1L1转录活性活性,辛伐他汀则增强NPC1L1转录活性,而-762T>C多态性不影响启动子活性及药物反应。展开更多
Type B Niemann-Pick disease is an autosomal recessive sphingolipidosis due to mutations in the sphingomyelin phosphodiesterase 1 gene （SMPD1）, Here we present molecular findings for two sibling patients. One mutatio...Type B Niemann-Pick disease is an autosomal recessive sphingolipidosis due to mutations in the sphingomyelin phosphodiesterase 1 gene （SMPD1）, Here we present molecular findings for two sibling patients. One mutation V36A due to c.107T〉C in exon 1 is a single nucleotide polymorphism and the other N522S due to c.1565 A〉G in exon 6 is a novel missense mutation. This non-fatal missense mutation leads to -20% residual lysosomal acid sphingomyelinase activity in vitro and only results in hepatosplenomegaly without neurologic involvement,展开更多
Niemann-Pick disease type C1 （NPC1）, caused by mutations of NPC1 gene, is an inherited lysosomal lipid storage disorder. Loss of functional NPC1 causes the accumulation of free cholesterol （FC） in endocytic organe...Niemann-Pick disease type C1 （NPC1）, caused by mutations of NPC1 gene, is an inherited lysosomal lipid storage disorder. Loss of functional NPC1 causes the accumulation of free cholesterol （FC） in endocytic organelles that comprised the characteristics of late endosomes and/or lysosomes. In this study we analyzed the pathogenic effect of 103 nsSNPs reported in NPC1 using computational methods. Rl186C, S940L, R958Q and I1061T mutations were predicted as most deleterious and disease associated with NPC1 using SIFT, Polyphen 2.0, PANTHER, PhD-SNP, Pmut and MUTPred tools which were also endorsed with previous in vivo experimental studies. To understand the atomic arrangement in 3D space, the native and disease associated mutant （Rl186C, S940L, R958Q and I1061T） structures were modeled. Quantitative structural and flexibility analysis was conceded to observe the structural consequence of prioritized disease associated mutations （R1186C, S940L, R958Q and I1061T）. Accessible surface area （ASA）, free folding energy （FFE） and hydrogen bond （NH bond） showed more flexibility in 3D space in mutant structures. Based on the quantitative assessment and flexibility analysis of NPC1 variants, I1061T showed the most deleterious effect. Our analysis provides a clear clue to wet laboratory scientists to understand the structural and functional effect of NPCI gene upon mutation.展开更多
目的研究姜黄素与高脂饮食小鼠胆囊胆固醇性结石关系及可能参与调控过程的因子。方法将C57BL6小鼠50只随机分为5组,其中一组予普通饲料,其余四组予高脂饮食饲料,同时灌喂不同剂量姜黄素[0、200、500、1 000 mg/(kg·d)],共4周。计...目的研究姜黄素与高脂饮食小鼠胆囊胆固醇性结石关系及可能参与调控过程的因子。方法将C57BL6小鼠50只随机分为5组,其中一组予普通饲料,其余四组予高脂饮食饲料,同时灌喂不同剂量姜黄素[0、200、500、1 000 mg/(kg·d)],共4周。计算小鼠胆囊结石成石率,收集小鼠血液、胆囊胆汁、胆囊、肝脏以及小肠。计算胆囊容积,称量肝脏质量,检测血液及胆汁胆固醇、甘油三酯等生化指标。用Realtime PCR及Western blotting方法分别检测小鼠小肠上皮NPC1L1及SREBP2 m RNA及蛋白表达。结果姜黄素能够降低小鼠胆囊结石的发生,大剂量姜黄素可降低胆囊结石发生率达60%,同时降低肝脏脂肪变性程度,降低小鼠血液胆固醇饱和度。同时,姜黄素可以抑制高脂饮食所致的NPC1L1 m RNA表达上调[(2.65±0.04)vs(2.06±0.07),(1.69±0.06),(1.33±0.05),P<0.01],且呈现剂量依赖关系,降低高脂饮食所致的NPC1L1蛋白高表达。姜黄素还可以抑制高脂饮食所致的SREBP2 m RNA表达上调[(1.34±0.08)vs(1.39±0.03),(1.19±0.01),(1.06±0.03),P<0.05],且呈现剂量依赖关系,降低高脂饮食所致的SREBP2蛋白表达上调。结论姜黄素能够降低高脂饮食小鼠胆囊结石的形成,NPC1L1和SREBP2可能参与了这个过程。展开更多
尼曼-匹克C1型类似蛋白1(Niemann-Pick type C1 Like 1, NPC1L1)是一种跨膜蛋白,是外源性胆固醇吸收的重要因子,在体内胆固醇代谢过程中发挥十分重要的作用。NPC1L1与多种脂质转运体共同影响着胆固醇的代谢。细胞核受体主要通过作用于NP...尼曼-匹克C1型类似蛋白1(Niemann-Pick type C1 Like 1, NPC1L1)是一种跨膜蛋白,是外源性胆固醇吸收的重要因子,在体内胆固醇代谢过程中发挥十分重要的作用。NPC1L1与多种脂质转运体共同影响着胆固醇的代谢。细胞核受体主要通过作用于NPC1L1启动子区域调控NPC1L1的表达,进而影响胆固醇的吸收,但其影响胆固醇吸收的具体机制还没有完全清楚。NPC1L1的表达受多种因子的调节。多不饱和脂肪酸通过甾体调节原件结合蛋白2(sterol regulatory element binding protein 2, SREBP2)途径下调NPC1L1的表达。姜黄素及鞘氨醇等也参与NPC1L1表达的调节。降脂药物依泽替米贝(ezetimibe)可通过降低NPC1L1的表达减少胆固醇的吸收从而降低血浆胆固醇的水平,同时对其它脂类代谢病也有一定的作用。本文对NPC1L1在结构、功能和调节方面的研究进展做一综述。展开更多
文摘Niemann-Pick C1 Like 1(NPC1L1)是肠道胆固醇吸收的关键蛋白,也是胆固醇吸收抑制剂依泽替米贝的分子作用靶点。NPC1L1的表达受一些核因子受体调控。敲除apoE-/-小鼠NPC1L1基因能显著抑制动脉粥样硬化的发生和发展,为动脉粥样硬化和冠状动脉性心脏病提供了新的治疗靶点。
文摘目的:研究尼曼匹克C1样1(Niemann Pick C1 like 1,NPC1L1)基因启动子-762T>C多态性的活性差异及药物对其的调节作用。方法:分别构建含NPC1L1基因-762T>C多态性的T/C等位基因启动子荧光酶报告基因,测定启动子活性以及不同药物处理后启动子活性的改变。结果:含T或C等位基因的NPC1L1基因启动子活性无统计学差异(P>0.05)。外源性胆固醇和依泽麦布均在高浓度抑制两种启动子活性,辛伐他汀则增强两种启动子活性(P<0.05),但两种基因型启动子之间对药物依泽麦布和辛伐他汀的反应无统计学差异(P>0.05)。结论:胆固醇和依泽麦布能抑制NPC1L1转录活性活性,辛伐他汀则增强NPC1L1转录活性,而-762T>C多态性不影响启动子活性及药物反应。
文摘目的观察肠道胆固醇吸收抑制剂依泽替米贝(ezetimibe)对RAW264.7细胞源性荷脂细胞脂质蓄积的影响并对其机制进行初步探讨。方法采用油红O染色、高效液相色谱法检测细胞内脂滴数量和细胞内脂质含量,Western blot对NPC1L1(Niemann-Pick type C1Like-1)进行定性和半定量检测。结果RAW264.7细胞中有NPC1L1蛋白表达。不同浓度(0、0.003、0.01和0.03mol.L-1)依泽替米贝预先孵育RAW264.7细胞24h或最佳浓度(0.03mol.L-1)预先孵育不同时间(0、6、12和24h)后,换50mg.L-1oxLDL继续孵育24h,结果显示不同浓度ezetimibe预先孵育后,细胞内脂滴数量与面积随着浓度的增加而逐渐减少;Ezetimibe预先孵育可减少细胞内脂质蓄积,并呈浓度和时间依赖性。其中0.03mol.L-1ezetimibe预先孵育24h组作用最明显,CE百分比较oxLDL单独孵育组减少了约47%±0.1%。结论小鼠源性巨噬细胞RAW264.7中存在NPC1L1蛋白表达;依泽替米贝能够减少RAW264.7细胞中NPC1L1蛋白表达;依泽替米贝抑制RAW264.7细胞中脂质蓄积。
文摘Type B Niemann-Pick disease is an autosomal recessive sphingolipidosis due to mutations in the sphingomyelin phosphodiesterase 1 gene （SMPD1）, Here we present molecular findings for two sibling patients. One mutation V36A due to c.107T〉C in exon 1 is a single nucleotide polymorphism and the other N522S due to c.1565 A〉G in exon 6 is a novel missense mutation. This non-fatal missense mutation leads to -20% residual lysosomal acid sphingomyelinase activity in vitro and only results in hepatosplenomegaly without neurologic involvement,
文摘Niemann-Pick disease type C1 （NPC1）, caused by mutations of NPC1 gene, is an inherited lysosomal lipid storage disorder. Loss of functional NPC1 causes the accumulation of free cholesterol （FC） in endocytic organelles that comprised the characteristics of late endosomes and/or lysosomes. In this study we analyzed the pathogenic effect of 103 nsSNPs reported in NPC1 using computational methods. Rl186C, S940L, R958Q and I1061T mutations were predicted as most deleterious and disease associated with NPC1 using SIFT, Polyphen 2.0, PANTHER, PhD-SNP, Pmut and MUTPred tools which were also endorsed with previous in vivo experimental studies. To understand the atomic arrangement in 3D space, the native and disease associated mutant （Rl186C, S940L, R958Q and I1061T） structures were modeled. Quantitative structural and flexibility analysis was conceded to observe the structural consequence of prioritized disease associated mutations （R1186C, S940L, R958Q and I1061T）. Accessible surface area （ASA）, free folding energy （FFE） and hydrogen bond （NH bond） showed more flexibility in 3D space in mutant structures. Based on the quantitative assessment and flexibility analysis of NPC1 variants, I1061T showed the most deleterious effect. Our analysis provides a clear clue to wet laboratory scientists to understand the structural and functional effect of NPCI gene upon mutation.
文摘目的研究姜黄素与高脂饮食小鼠胆囊胆固醇性结石关系及可能参与调控过程的因子。方法将C57BL6小鼠50只随机分为5组,其中一组予普通饲料,其余四组予高脂饮食饲料,同时灌喂不同剂量姜黄素[0、200、500、1 000 mg/(kg·d)],共4周。计算小鼠胆囊结石成石率,收集小鼠血液、胆囊胆汁、胆囊、肝脏以及小肠。计算胆囊容积,称量肝脏质量,检测血液及胆汁胆固醇、甘油三酯等生化指标。用Realtime PCR及Western blotting方法分别检测小鼠小肠上皮NPC1L1及SREBP2 m RNA及蛋白表达。结果姜黄素能够降低小鼠胆囊结石的发生,大剂量姜黄素可降低胆囊结石发生率达60%,同时降低肝脏脂肪变性程度,降低小鼠血液胆固醇饱和度。同时,姜黄素可以抑制高脂饮食所致的NPC1L1 m RNA表达上调[(2.65±0.04)vs(2.06±0.07),(1.69±0.06),(1.33±0.05),P<0.01],且呈现剂量依赖关系,降低高脂饮食所致的NPC1L1蛋白高表达。姜黄素还可以抑制高脂饮食所致的SREBP2 m RNA表达上调[(1.34±0.08)vs(1.39±0.03),(1.19±0.01),(1.06±0.03),P<0.05],且呈现剂量依赖关系,降低高脂饮食所致的SREBP2蛋白表达上调。结论姜黄素能够降低高脂饮食小鼠胆囊结石的形成,NPC1L1和SREBP2可能参与了这个过程。
基金supported by the National Natural Science Foundation of China (No.81071416)
文摘尼曼-匹克C1型类似蛋白1(Niemann-Pick type C1 Like 1, NPC1L1)是一种跨膜蛋白,是外源性胆固醇吸收的重要因子,在体内胆固醇代谢过程中发挥十分重要的作用。NPC1L1与多种脂质转运体共同影响着胆固醇的代谢。细胞核受体主要通过作用于NPC1L1启动子区域调控NPC1L1的表达,进而影响胆固醇的吸收,但其影响胆固醇吸收的具体机制还没有完全清楚。NPC1L1的表达受多种因子的调节。多不饱和脂肪酸通过甾体调节原件结合蛋白2(sterol regulatory element binding protein 2, SREBP2)途径下调NPC1L1的表达。姜黄素及鞘氨醇等也参与NPC1L1表达的调节。降脂药物依泽替米贝(ezetimibe)可通过降低NPC1L1的表达减少胆固醇的吸收从而降低血浆胆固醇的水平,同时对其它脂类代谢病也有一定的作用。本文对NPC1L1在结构、功能和调节方面的研究进展做一综述。
