催产素对3T3-L1脂肪细胞糖脂代谢的影响

目的观察催产素对3T3-L1脂肪细胞糖脂代谢的影响。方法 3T3-L1前脂肪细胞体外培养,并诱导其分化成熟为脂肪细胞。研究催产素对脂肪细胞葡萄糖消耗量以及三酰甘油、游离脂肪酸和甘油的影响。采用实时荧光定量PCR法检测糖脂代谢相关基因GLUT-1、GLUT-4、ATGT、HSL的mRNA表达。结果与对照组比较,催产素20、50、100μg/m L组葡萄糖消耗量有所增加,且表现出剂量相关。催产素组较对照组的三酰甘油降低,而甘油和游离脂肪酸增高。催产素50μg/m L组中脂代谢相关基因HSL表达明显高于对照组,糖代谢相关基因GLUT-4 mRNA表达水平增加。结论催产素处理可减少3T3-L1细胞脂质合成、增加脂质分解作用,并可明显改善脂质积聚。     

葛根调节脂肪细胞糖脂代谢改善胰岛素抵抗的研究

该实验通过检测葛根对胰岛素抵抗(IR)3T3-L1脂肪细胞葡萄糖消耗,甘油三脂(TG)含量及PPARγ,ADPN,GLUT4,LPL,FABP4,FASn表达量的影响来探讨葛根调节糖脂代谢改善脂肪IR的作用机制。先采用3T3-L1前脂细胞诱导分化的成熟脂肪细胞,地塞米松诱导建立IR模型,将脂肪细胞分为正常组,IR模型组,罗格列酮阳性组,低、中、高剂量葛根含药血清组,以葡萄糖氧化酶-过氧化物酶(glucose oxidase-peroxidase,GOD-POD)法检测细胞培养液葡萄糖含量和甘油磷酸氧化酶(glycerol phosphate oxidase,GPO-POD)法测定胞内TG含量,荧光定量q PCR检测PPARγ,ADPN,GLUT4,LPL,FABP4(a P2),FASn基因mRNA水平。结果显示1μmol·L~(-1)地塞米松作用3T3-L1脂肪细胞96 h,与正常组比较,模型组葡萄糖消耗量降低(P0.01),胞内TG含量增加(P0.01),由此确认建立IR模型;与IR组比较,葛根含药血清干预IR细胞24 h葡萄糖消耗量上升(P0.01),胞内TG含量降低(P0.01),中、高剂量葛根含药血清组升高PPARγ,ADPN和GLUT4表达(P0.01),PPARγ与后两者基因表达呈现一致性。脂代谢相关基因检测结果显示仅高剂量葛根含药血清显著升高LPL表达(P0.05);各剂量葛根含药血清下调FABP4表达(P0.01);中、高剂量的葛根含药血清上调FASn基因表达(P0.01)。该实验表明葛根提高IR-3T3-L1脂肪细胞对葡萄糖的摄取能力,降低细胞内TG积聚,干预多个重要糖脂代谢基因,推测以PPARγ为中心多靶点调节糖脂代谢改善脂肪IR。     

梓醇与小檗碱及其配伍对胰岛素抵抗3T3-L1脂肪细胞的影响

目的观察梓醇与小檗碱及其配伍对地塞米松诱导的胰岛素抵抗3T3-L1脂肪细胞葡萄糖消耗、转运及这一过程中过氧化物体增殖物激活受体(PPAR-γ)mRNA表达的影响。方法采用地塞米松诱导胰岛素抵抗细胞模型,分别给予罗格列酮、小檗碱、梓醇、梓醇 小檗碱进行干预,以葡萄糖氧化酶法检测培养液中葡萄糖消耗量,以2-脱氧-[3H]-D-葡萄糖摄入法观察葡萄糖的转运率,以RT-PCR检测PPAR-γmRNA的表达。结果含或不含10nmol/L胰岛素的条件下,梓醇、小檗碱及其配伍组胰岛素抵抗脂肪细胞的葡萄糖消耗量和转运率较模型组明显改善(P<0.05、0.01),配伍组效应优于梓醇、小檗碱单药组(P<0.05、0.01);且小檗碱组及配伍组PPAR-γmR-NA的表达降低(P<0.05、0.01)。结论梓醇、小檗碱均能增加葡萄糖消耗和转运,改善胰岛素抵抗,其作用不依赖胰岛素的存在,且小檗碱及两药配伍还能下调脂肪细胞PPAR-γmRNA的表达水平,提示梓醇、小檗碱改善胰岛素抵抗的作用机制可能与罗格列酮不同。     

葛根素对3T3-L1前脂肪细胞分化及胰岛素抵抗模型糖代谢的影响

目的:研究葛根素(Pur)对3T3-L1前脂肪细胞增殖、分化的影响,以及在胰岛素抵抗状态下对3T3-L1脂肪细胞葡萄糖代谢的影响。方法:不同浓度Pur干预3T3-L1细胞,MTT法检测其对细胞增殖的影响;油红O染色法检测其对前脂肪细胞分化过程及成脂的影响;地塞米松诱导3T3-L1细胞建立胰岛素抵抗模型,用不同浓度Pur进行干预,测定细胞的葡萄糖消耗量。结果:Pur(3～300μmol/L)对3T3-L1前脂肪细胞增殖无明显影响;Pur(30～300μmol/L)促进3T3-L1细胞的分化成脂,增加胰岛素抵抗状态下3T3-L1脂肪细胞的葡萄糖代谢,且具有量效关系。结论:Pur能够促进3T3-L1细胞的分化成脂,增加胰岛素抵抗状态下3T3-L1脂肪细胞的葡萄糖代谢,改善胰岛素抵抗。     

Astragaloside IV attenuates lipolysis and improves insulin resistance induced by TNFalpha in 3T3-L1 adipocytes

Increased circulating free fatty acid (FFA) concentrations have been demonstrated to potentially link obesity, insulin resistance and cardiovascular diseases. Astragaloside IV (AS-IV) is a saponin which is widely used in traditional Chinese medicine to treat type 2 diabetes and cardiovascular diseases. The purpose of the present study was to examine the effects of AS-IV on the lipolysis and insulin resistance induced by tumor necrosis factor-alpha (TNFalpha) in cultured 3T3-L1 adipocytes. TNFalpha promotes lipolysis in mammal adipocytes via the mitogen activated protein kinase (MAPK) family resulting in reduced expression/function of perilipin. Application of AS-IV inhibited TNFalpha-induced accelerated lipolysis in a dose-dependent manner, which was compatible with suppressed phosphorylation of ERK1/2 and reversed the downregulation of perilipin. Moreover, TNFalpha induced downregulation of key enzymes in lipogenesis, including LPL, FAS and GPAT, were also attenuated by AS-IV. Further studies showed that AS-IV improved TNFalpha-induced insulin resistance in 3T3-L1 adipocytes. This study provides the first direct evidence of the antilipolytic action of AS-IV in adipocytes, which may allow this agent to decrease the circulating FFA levels, thus increase insulin sensitivity and treat cardiovascular diseases.     

齐墩果酸抑制3T3-L1脂肪细胞脂质堆积

目的：从57个中药化合物筛选具有抑制3T3-L1脂肪细胞脂质堆积的活性成分。方法：3T3-L1细胞培养至融合后用化合物进行干预,并诱导使其分化,采用高内涵影像方法检测细胞中脂滴含量。对发现的活性成分用ToxInsight体外毒性检测方法评价其肝毒性。结果：发现并验证活性成分齐墩果酸（OA）具有显著的脂质形成抑制作用,其半数抑制浓度（IC50）为14.5 μmol·L-1,且在60 μmol·L-1测试浓度范围内对HepG2 细胞无显著损伤作用。结论：OA 具有显著的降低脂质形成作用,是潜在的降脂候选化合物。     

Effects of capsaicin on lipid catabolism in 3T3-L1 adipocytes

Capsaicin (8-methyl-N-vanillyl-6-nonenamide) is a pungent ingredient of red peppers, and has been reported to reduce body weight gain and adiposity in rodents. The present study investigated the effects of capsaicin on lipid catabolism in differentiated 3T3-L1 adipocytes. Capsaicin decreased the intracellular lipid content in a concentration-dependent manner. The release of glycerol into the medium was increased by the addition of capsaicin. The mRNA levels of genes involved in lipid catabolism such as hormone sensitive lipase (HSL), carnitine palmitoyl transferase-Iα (CPTI-α) and uncoupling protein 2 (UCP2) were up-regulated significantly. These results suggest that capsaicin exerts its lipolytic action by increasing the hydrolysis of triacylglycerol in adipocytes, and that these effects are mediated at least partially by regulation of the expression of multiple genes that are involved in the lipid catabolic pathway, such as HSL and CPT-Iα, and those involved in thermogenesis such as UCP2.     

代综方对不同方式诱导3T3-L1脂肪细胞胰岛素抵抗的改善作用

目的:探讨中药代综方(DZF)对不同方式诱导的脂肪细胞胰岛素抵抗(IR)的改善作用.方法:鸡尾酒式联合诱导3T3-L1前脂肪细胞分化成熟,分别采用棕榈酸(PA),高浓度葡萄糖(HG),地塞米松(DEX)诱导建立IR模型.不同质量浓度DZF提取物(含生药质量浓度2.0,0.5,0.1 g·L-1)干预24 h,另设模型组...     

葛根素改善3T3-L1 脂肪细胞葡萄糖摄取和胰岛素抵抗

目的：探讨葛根素对3T3-L1脂肪细胞胰岛素抵抗（Insulin resistance,IR）模型葡萄糖利用和IR的影响及其作用机制。方法：诱导分化3T3-L1前脂肪细胞为成熟脂肪细胞;用地塞米松诱导建立3T3-L1脂肪细胞胰岛素抵抗模型。成功建立模型后,将实验分为5组进行,分别为空白组、模型组、葛根素10 mg·L-1组、葛根素100 mg·L-1组、葛根素200 mg·L-1组;其中,空白组不建模,模型组建模但不用药物干预,各葛根素组为在建立模型后,给予不同浓度的葛根素干预。用葛根素干预24小时后,以葡萄糖氧化酶-过氧化物酶法测定细胞培养基中葡萄糖浓度,四甲基偶氮唑蓝[3-(4,5-dimehyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazoliumbromide, MTT]比色法检测细胞增值活力;然后分别用Real time PCR、Western Blot 方法检测细胞葡萄糖转运蛋白4（glucosetransporter type 4, GLUT4）、磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase, PI3K）、蛋白激酶B（protein kinaseB, PKB/AKT）、腺苷酸活化蛋白激酶（adenosine monophosphate activated protein kinase, AMPK）、过氧化物酶体增殖物激活受体γ（peroxisome proliferator -activated receptor gamma, PPARγ）mRNA基因和蛋白表达。结果：葛根素干预后,细胞培养基中葡萄糖浓度均较模型组降低（P < 0.01）;GLUT4、PI3K、AKT、AMPK、PPARγ mRNA水平及蛋白表达较模型组均增加（P < 0.05）。结论：葛根素能增加3T3-L1脂肪细胞胰岛素抵抗模型葡萄糖利用、改善IR,其作用机制可能与上调P13K、AKT、AMPK、PPARγ基因和蛋白表达,进而增加葡萄糖运转有关。     

小檗碱与梓醇及其配伍对胰岛素抵抗3T3-L1脂肪细胞葡萄糖转运子4蛋白及C-Cb1相关蛋白表达的影响

目的观察梓醇与小檗碱及其配伍对胰岛素抵抗3T3-L1脂肪细胞葡萄糖消耗及这一过程中葡萄糖转运子4(Glut4)和C-Cb1相关蛋白(CAP)表达的影响。方法采用高糖联合高胰岛素诱导3T3-L1脂肪细胞产生胰岛素抵抗(IR),分别给予小檗碱、梓醇、小檗碱 梓醇、盐酸罗格列酮进行干预,以葡萄糖氧化酶法检测培养液中葡萄糖消耗量,以Western Blotting法检测Glut4和CAP蛋白的表达。结果与模型组相比,小檗碱能增加培养液中葡萄糖的消耗,但对Glut4蛋白的表达无影响;梓醇、小檗碱 梓醇均能显著增加培养液中葡萄糖的消耗,并使细胞中Glut4蛋白的表达增强,且小檗碱 梓醇组的效应优于梓醇组及小檗碱组;与模型组相比,小檗碱与梓醇及其配伍对CAP的表达没有显著性影响。结论小檗碱、梓醇及其配伍能改善IR 3T3-L1脂肪细胞的胰岛素活性,其作用机制与罗格列酮不同。     

胰岛素抵抗3T3-L1脂肪细胞模型及中药有效成分的干预作用

高糖高胰岛素、游离脂肪酸、炎症因子、地塞米松、金属铬、雌激素、葡糖胺等均可诱导3T3-L1脂肪细胞建立胰岛素抵抗模型,且具有稳定、可靠、易于重复等优点。目前已证实小檗碱+梓醇、小檗碱、熊果酸、西洋参茎叶总皂苷及其他活性成分、蒲黄总黄酮、灵芝多糖、附子多糖、地黄寡糖、积雪草酸、白藜芦醇、葛根素等中药有效成分具有改善胰岛素抵抗及调节糖代谢作用。     

3,4-Oxo-isopropylidene-shikimic acid promotes adiopkine expression during murine 3T3-L1 fibroblast differentiation into adipocytes

Objective3,4-Oxo-isopropylidene-shikimic acid (ISA), a derivative of shikimic acid, has exhibited ameliorative effect on cognitive impairment in experimental animal models of dementia. This study investigated the effect of ISA on lipid accumulation and adipokine secretion during differentiation of 3T3-L1 fibroblasts to adipocytes.Methods3T3-L1 cells were cultured and treated with ISA (50–800 μM) from days 3–8. Lipid accumulation and triglyceride content were measured. Gene expression of adipokines (adiponectin, leptin, and resistin), CCAAT/enhancer binding protein (C/EBP) β, C/EBP α and peroxisome proliferator-activated receptor γ (PPAR γ) and PPAR target genes, including adipocyte fatty acid binding protein (aP2) and fatty acid synthase (FAS) were investigated.ResultsISA promoted 3T3-L1 fibroblast differentiation to adipocytes and increased triglyceride content by 26%. On mechanistic levels, ISA increased expressions of C/EBP β, PPAR γ, C/EBP α, aP2 and FAS. Moreover, ISA stimulated expressions of adipokines secreted by adipocytes, including adiponectin, leptin, and resistin.ConclusionsThese findings demonstrated that ISA promoted adipogenesis by up-regulating expressions of C/EBP β, PPAR γ, C/EBP α, aP2 and FAS, and also stimulated adipokines during adipocyte differentiation. Further study should clarify the relationship between stimulation of adipokines and cognitive enhancing effect of ISA.     

含左归丸鼠血清对3T3-L1前脂肪细胞芳香化酶mRNA表达的影响

目的：观察含左归丸鼠血清对3T3-L1前脂肪细胞芳香化酶 mRNA 表达的影响。方法：将雌性育龄期大鼠分为正常组20只、假手术组17只、模型组8只、阳性对照组11只、左归丸高剂量组（左高组）6只、左归丸低剂量组（左低组）9只,连续灌胃11周,分离血清检测 E2;以2％各类鼠血清加入培养液培养分化中的3T3-L1前脂肪细胞,48 h 后测细胞上清液E2和细胞芳香化酶 mRNA 表达。结果：与模型组比较,左高组、左低组给药前后体重有升高趋势,但无明显统计学意义（P＞0.05）;左高组、左低组鼠血清 E2值较模型组无明显统计学意义（P ＞0.05）;培养3T3-L1前脂肪细胞48h 后,细胞上清液 E2值左高组（9.40±3.61）pg／mL、左低组（8.94±2.91）pg／mL 比模型组（3.99±0.76）pg／mL 略有升高;模型组（0.39）芳香化酶 mRNA 表达（P450arom／GAPDH）水平高于左高组（0.25）。结论：左归丸对去势雌鼠血清 E2和体重无明显影响,可增强3T3-L1前脂肪细胞的芳香化酶活性。     

小檗碱对3T3-L1前脂肪细胞及其胰岛素抵抗模型的影响

目的:探查小檗碱的体外安全浓度及其对脂肪细胞的抗胰岛素抵抗(IR)活性。方法:3T3-L1前脂肪细胞分为空白组和小檗碱不同浓度(0.001~10μmol·L-1)组,48 h后MTT法测定各组的A值。成熟的3T3-L1脂肪细胞分为空白组和IR模型组,模型组细胞经10μmol·L-1胰岛素(Ins)诱导24,48 h。检测各组培养液和细胞葡萄糖含量。检测IR模型细胞经小檗碱不同浓度(0.000 1~1μmol·L-1)干预24,48 h后的培养液中葡萄糖含量。结果:小檗碱0.01~1μmol·L-13个浓度和10μmol·L-1作用的前脂肪细胞A值均显著升高或降低(P0.05,P0.01);经Ins诱导24,48 h的培养液和细胞葡萄糖含量均显著增加或减少(P0.01);作用24 h的小檗碱0.01~1μmol·L-1和作用48 h的0.000 1~1μmol·L-1各浓度组培养液中的葡萄糖量均显著减少(P0.01)。结论:小檗碱的体外细胞安全浓度较低,低浓度小檗碱对脂肪细胞即具有显著的抗IR活性。     

小檗碱对3T3-L1脂肪细胞脂联素表达的影响

目的:探讨小檗碱和胰岛素对3T3-L1脂肪细胞脂联素表达的影响。方法:检测分别经小檗碱及胰岛素处理后3T3 L1脂肪细胞脂联素表达水平改变,以β actin为内对照,半定量法RT PCR测定脂联素mRNA表达。结果:经小檗碱(10μmol·L^-1)干预后3T3 L1脂肪细胞脂联素表达显著增加(P<0.05),加入胰岛素后脂联素的表达受到抑制,高浓度胰岛素有显著抑制作用(P<0.05)。结论:体外实验中,小檗碱使3T3-L1脂肪细胞脂联素的表达增加,而胰岛素可抑制小檗碱增加脂联素表达的作用。     

Effect of ginsenosides Rg3 and Re on glucose transport in mature 3T3-L1 adipocytes

Ginsenosides, the active component of Panax ginseng, have been shown to evidence a variety of biological activities associated with hyperglycemia, obesity and type 2 diabetes mellitus. This study evaluated the effects of the ginsenosides, Rg3 and Re, on glucose uptake and the glucose transport system in mature 3T3‐L1 cells. The results demonstrated that the glucose uptake of ginsenosides Rg3 and Re at concentrations of 1–10 µM significantly increased by approximately ～10% and ～12%, respectively. Furthermore, the glucose transporter 4 (GLUT4) mRNA expression of ginsenosides Rg3 and Re at 10 µM was increased by approximately ～1.73 and 1.43 fold, respectively. It was further confirmed in a series of experiments that ginsenosides Rg3 and Re stimulated the mRNA expression of insulin receptor substrate (IRS‐1) and the expression of phosphatidylinositol 3‐kinase (PI3K)‐110α protein, which is involved in downstream events in the insulin signaling pathway. These findings demonstrate that ginsenosides Rg3 and Re may stimulate glucose uptake via the PI3K pathways involving IRS‐1. Further, our results suggest that both of these ginsenosides might prove useful as effective antidiabetic and antihyperglycemic agents. Copyright © 2010 John Wiley & Sons, Ltd.     

芦丁促进3T3-L1前脂肪细胞棕色化及其机制

目的：观察芦丁对3T3-L1前脂肪细胞棕色化效应的影响,并探讨其机制。方法：细胞增殖与活性检测-8(CCK-8法检测不同浓度芦丁(3.125、6.25、12.5、25、50、100、200μmol·L^-1)对3T3-L1细胞活性影响,蛋白免疫印迹法(Western blot)检测不同浓度芦丁(12.5、25、50μmol·L^-1)对脂肪细胞产热相关蛋白解偶联蛋白1(UCP1)、PR结构域蛋白16(PRDM16)、过氧化物酶体增殖物激活受体γ辅助激活因子-1α(PGC-1α)表达的影响。确定芦丁最佳浓度后,油红O染色观察芦丁对脂肪细胞中脂滴生成的影响,Western blot检测线粒体生物合成标志性蛋白核呼吸因子1(NRF1)、核呼吸因子2(NRF2)和线粒体转录因子A(TFAM)的表达。结果：与空白组比较,200μmol·L^-1芦丁显著抑制3T3-L1细胞活性(P<0.01);在12.5、25、50μmol·L^-1浓度下,50μmol·L^-1芦丁显著促进产热蛋白(UCP1...