Perilipin1(PLIN1)is a major phosphorylated protein that specifically coats the surface of neutral lipid droplets(LDs)in adipocytes and plays a crucial role in regulating the accumulation and hydrolysis of triacylglyce...Perilipin1(PLIN1)is a major phosphorylated protein that specifically coats the surface of neutral lipid droplets(LDs)in adipocytes and plays a crucial role in regulating the accumulation and hydrolysis of triacylglycerol(TG).Mammalian studies have shown that Plin1 gene transcription is mainly regulated by peroxisome proliferator-activated receptorgamma(PPARγ),the master regulator of adipogenesis.However,the regulatory mechanism of the chicken Plin1(c Plin1)gene is poorly understood.The present study aimed to investigate whether Plin1 is regulated by PPARγin chickens and identify its exact molecular mechanism.Reporter gene and expression assays showed that PPARγ2,but not PPARγ1,activated(P<0.01)the cPlin1 gene promoter.An electrophoretic mobility shift assay and mutational analysis revealed that PPARγ2 bound to a special site in the cPlin1 gene promoter to enhance its expression.In summary,our results show that PPARγpromotes the expression of the cPlin1 gene and that PPARγ2 is the main regulatory isoform.展开更多
目的探讨雌激素对h ASCs成脂分化过程中脂滴相关特异基因DNA断裂因子相似蛋白C(The cell death-inducing DNA fragmentation 45-like effector,CIDEc)、脂滴包被蛋白(Perilipin1,Plin1)m RNA表达的影响。方法对3例患者行脂肪抽吸术,并进...目的探讨雌激素对h ASCs成脂分化过程中脂滴相关特异基因DNA断裂因子相似蛋白C(The cell death-inducing DNA fragmentation 45-like effector,CIDEc)、脂滴包被蛋白(Perilipin1,Plin1)m RNA表达的影响。方法对3例患者行脂肪抽吸术,并进行h ASCs培养及传代,鉴定表面标志物;经典鸡尾酒法诱导其成脂,不同浓度(10-8、10-7、10-6mol/L)17β-雌二醇(17β-E2)作用于细胞,分别于24 h、4、7、11d收获细胞,用RT-PCR检测CIDEc、Plin1 m RNA表达情况。结果各组细胞经药物作用后,脂肪细胞中CIDEc及Plin1 m RNA的表达水平随着17β-E2浓度的升高而下降;而随着作用时间的延长,CIDEc及Plin1 m RNA表达水平分别于7 d和4 d达到高峰期,其后逐渐下降。当17β-E2浓度为10-6mol/L时,细胞中CIDEc及Plin1 m RNA的表达水平最低,差异有统计学意义(P<0.01);而加入雌激素受体阻滞剂ICI182780的一组细胞,雌激素对CIDEc及Plin1 m RNA表达的抑制作用明显减弱。结论 17β-E2能够显著降低脂肪细胞分化过程中CIDEc及Plin1的m RNA表达,并可能通过这一途径抑制脂肪细胞内脂滴的增殖。展开更多
基金supported by grants from the Youth Innovative Talents Program and Characteristic Innovative Talents Program of Regular Institutions of Higher Education of Guangdong Province(2018KQNCX212,2020KTSCX101)the Scientific Research Project of Traditional Chinese Medicine Bureau of Guangdong Province(20222122)+3 种基金the Excellent Youth Science Research Program of the Education Department of Hunan Province(14B183)Bureau of Education of Guangzhou Municipality(202032801)the Plan on Enhancing Scientific Research in Guangzhou Medical Universitythe Open Research Funds(2021)and the Funds of Selected Project(2022)from GMU-GIBH Joint School of Life Sciences,Guangzhou Medical University。
文摘目的oxLDL可上调Plin2的表达,进而促进泡沫细胞的形成,LOX1是oxLDL的受体。本文探讨Plin2与LOX1在动脉粥样硬化发生发展过程中的关系。方法从GEO数据库中下载GSE43292,分析Plin2、LOX1的表达及Plin2、LOX1与NF-κB信号通路的相关性。采用oxLDL处理的RAW264.7细胞作为动脉粥样硬化的细胞模型进行研究,蛋白质免疫印迹法检测细胞中Plin2、LOX1和p-p65的表达,荧光中性脂质染料BODIPY 493/503染色法检测细胞内脂滴。结果通过分析GSE43292数据发现,Plin2、LOX1在颈动脉粥样硬化斑块中的表达显著高于颈动脉邻近组织。oxLDL处理RAW264.7细胞24 h后,Plin2与LOX1的表达、细胞内脂滴明显增加。过表达Plin2的细胞中LOX1表达升高;当用oxLDL孵育过表达Plin2的细胞后,LOX1的水平升高更为显著;但在没有oxLDL处理的情况下,敲减Plin2对细胞内LOX1的表达没有影响。基因集富集分析(gene set enrichment analysis,GSEA)结果显示,在动脉粥样硬化中,Plin2和LOX1的表达与NF-κB的活化呈正相关。此外,尽管采用oxLDL处理细胞,NF-κB抑制剂JSH-23预处理仍可显著降低Plin2与LOX1的表达、细胞内的脂质积聚,过表达Plin2后,JSH-23亦能显著抑制oxLDL孵育的细胞中Plin2和LOX1的表达。结论Plin2可通过上调LOX1的表达促进细胞内脂质积聚,参与动脉粥样硬化,这一过程至少部分是通过激活NF-κB通路实现的。
基金supported by the National Natural Science Foundation of China(31201796 and 32072704)the China Agriculture Research System of MOF and MARA(CARS-41)the Natural Science Foundation of Heilongjiang Province,China(LH2020C017)。
文摘Perilipin1(PLIN1)is a major phosphorylated protein that specifically coats the surface of neutral lipid droplets(LDs)in adipocytes and plays a crucial role in regulating the accumulation and hydrolysis of triacylglycerol(TG).Mammalian studies have shown that Plin1 gene transcription is mainly regulated by peroxisome proliferator-activated receptorgamma(PPARγ),the master regulator of adipogenesis.However,the regulatory mechanism of the chicken Plin1(c Plin1)gene is poorly understood.The present study aimed to investigate whether Plin1 is regulated by PPARγin chickens and identify its exact molecular mechanism.Reporter gene and expression assays showed that PPARγ2,but not PPARγ1,activated(P<0.01)the cPlin1 gene promoter.An electrophoretic mobility shift assay and mutational analysis revealed that PPARγ2 bound to a special site in the cPlin1 gene promoter to enhance its expression.In summary,our results show that PPARγpromotes the expression of the cPlin1 gene and that PPARγ2 is the main regulatory isoform.
文摘目的探讨雌激素对h ASCs成脂分化过程中脂滴相关特异基因DNA断裂因子相似蛋白C(The cell death-inducing DNA fragmentation 45-like effector,CIDEc)、脂滴包被蛋白(Perilipin1,Plin1)m RNA表达的影响。方法对3例患者行脂肪抽吸术,并进行h ASCs培养及传代,鉴定表面标志物;经典鸡尾酒法诱导其成脂,不同浓度(10-8、10-7、10-6mol/L)17β-雌二醇(17β-E2)作用于细胞,分别于24 h、4、7、11d收获细胞,用RT-PCR检测CIDEc、Plin1 m RNA表达情况。结果各组细胞经药物作用后,脂肪细胞中CIDEc及Plin1 m RNA的表达水平随着17β-E2浓度的升高而下降;而随着作用时间的延长,CIDEc及Plin1 m RNA表达水平分别于7 d和4 d达到高峰期,其后逐渐下降。当17β-E2浓度为10-6mol/L时,细胞中CIDEc及Plin1 m RNA的表达水平最低,差异有统计学意义(P<0.01);而加入雌激素受体阻滞剂ICI182780的一组细胞,雌激素对CIDEc及Plin1 m RNA表达的抑制作用明显减弱。结论 17β-E2能够显著降低脂肪细胞分化过程中CIDEc及Plin1的m RNA表达,并可能通过这一途径抑制脂肪细胞内脂滴的增殖。
