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.     

Antiobesity effect of baicalin involves the modulations of proadipogenic and antiadipogenic regulators of the adipogenesis pathway

In this study, the antiobesity effects of baicalin, 5,6‐dihydroxyflavone‐7‐glucuronic acid, were characterized using an in vitro system of adipogenesis, i.e. fat cell formation. Baicalin‐treatment of 3T3‐L1 preadipocytes was shown to inhibit triglyceride accumulation and lipid droplet formation during induced adipogenesis. Microarray analyses showed that baicalin modulated the expression of genes located in pathways such as adipogenesis, cholesterol biosynthesis, focal adhesion and others. In the adipogenesis pathway, treatment with baicalin significantly down‐regulated terminal differentiation markers of adipocytes including fatty acid binding protein 4. The effects of baicalin on the core part of the adipogenesis pathway, however, were paradoxical; the expression levels of CCAAT/enhancer binding protein (C/EBP)β and C/EBPδ were up‐regulated, while the expression levels of the peroxisome proliferator‐activated receptor (PPAR)γ and C/EBPα were down‐regulated. The antiadipogenic mechanisms of baicalin can be explained by its effects on the upstream part of adipogenesis pathway; baicalin not only up‐regulates the antiadipogenic regulators, C/EBPγ, C/EBP homologous protein and Kruppel‐like factor (KLF)2, but also down‐regulates the proadipogenic regulator, KLF15. The overall effects of baicalin on these upstream regulators of adipogenesis were antiadipogenic, resulting in the down‐regulation of downstream genes and the inhibition of cellular fat accumulation. Copyright © 2009 John Wiley & Sons, Ltd.     

小檗碱对3T3-L1前脂肪细胞增殖及分化相关基因PPARγ、C/EBPα mRNA和蛋白表达的影响

目的 探讨小檗碱对脂肪细胞增殖、分化的影响及其机制。     

Effect of mollugin on apoptosis and adipogenesis of 3T3-L1 preadipocytes

The effect of mollugin, isolated from the roots of Rubia cordifolia L., on cell viability, apoptosis and adipogenesis in 3T3‐L1 preadipocytes was investigated. The inhibitory effect of mollugin (40–60 µm ) on cell viability was more significant in differentiated adipocytes than in 3T3‐L1 preadipocytes. In 3T3‐L1 cells, the cytotoxicity of mollugin was accompanied by apoptotic events including mitochondrial membrane potential (Δψm) loss and activation of caspase‐9, ‐3 and ‐7, leading to PARP degradation. Although the presence of 20 µm mollugin during induced adipocytic differentiation of 3T3‐L1 cells for 6 days failed to affect the cell viability, it could almost completely abrogate the differentiation‐associated morphology change and intracellular lipid accumulation. A similar level of inhibition was observed, when 20 µm mollugin was present during the early stage (D0–D2) of the differentiation period. In addition, the expression of C/EBPα, PPARγ1 and PPARγ2 was significantly down‐regulated. The presence of 20 µm mollugin during either middle stage (D2–D4) or late stage (D4–D6) of the differentiation period, however, caused the inhibition to a lesser extent. These results indicated that mollugin at high concentrations (40–60 µm ) exerted cytotoxicity via inducing apoptosis, whereas mollugin at a low concentration (20 µm ) suppressed adipocytic differentiation without exerting cytotoxicity in 3T3‐L1 preadipocytes. Copyright © 2010 John Wiley & Sons, Ltd.     

黄连素的临床新用

黄连素是一种常用的广谱抗菌药物。本文作者总结整理了近年来黄连素在治疗消化性溃疡等八种疾病的新用途,在临床上有一定的借鉴作用。     

Berberine inhibits SREBP‐1‐related clozapine and risperidone induced adipogenesis in 3T3‐L1 cells

Weight gain is a common and potentially serious complication associated with the treatment of second generation antipsychotics such as clozapine and risperidone. Increased peripheral adipogenesis via the SREBP‐1 pathway could be one critical mechanism responsible for antipsychotic drug‐induced weight gain. Berberine, a botanic alkaloid, has been shown in our previous studies to inhibit adipogenesis in cell and animal models. MTT was used to determine the cytotoxic effects of clozapine and risperidone in combination with berberine. Differentiation of 3T3‐L1 cells was monitored by Oil‐Red‐O staining and the expression of SREBP‐1 and related proteins was determined by real‐time RT‐PCR and western blotting. The results showed that neither clozapine nor risperidone, alone or in combination with berberine had significant effects on cell viability. Eight days treatment with 15 μm clozapine increased adipogenesis by 37.4% and 50 μm risperidone increased adipogenesis by 26.5% during 3T3‐L1 cell differentiation accompanied by increased SREBP‐1, PPARγ, C/EBPα, LDLR and Adiponectin gene expression. More importantly, the addition of 8 μm berberine diminished the induction of adipogenesis almost completely accompanied by down‐regulated mRNA and protein expression levels of SREBP‐1‐related proteins. These encouraging results may lead to the use of berberine as an adjuvant to prevent weight gain during second generation antipsychotic medication. Copyright © 2010 John Wiley & Sons, Ltd.     

Antiobesity Effects of a Sulfur Compound Thiacremonone Mediated via Down-regulation of Serum Triglyceride and Glucose Levels and Lipid Accumulation in the Liver of db/db Mice

Garlic is widely used as a spice. Garlic extracts exert anticancer and antiinflammatory effects, but its antiobesity efficacy studies have produced conflicting results. The antiobesity effects of thiacremonone, a sulfur compound isolated from garlic, was evaluated in obese db/db mice. Thiacremonone was orally administrated to mice for 3 weeks. The thiacremonone‐treated db/db mice showed a loss of body weight and decrease in blood triglyceride and glucose levels compared with the control mice. Histological analysis further revealed that thiacremonone significantly decreased lipid accumulation in the fatty livers of treated db/db mice. It was observed that GLUT‐4 expression and glucose uptake were up‐regulated by thiacremonone in 3T3‐L1 adipocytes. Thiacremonone treatment also suppressed expression levels of acetyl‐CoA carboxylase (ACC) and fatty acid synthase (FAS), which are involved in lipid metabolism, in the liver of db/db mice. In addition, thiacremonone enhanced peroxisome proliferator‐activated receptor γ (PPARγ) expression in the fatty liver. Taken together, these results suggest that thiacremonone may play a vital role in improving the management of obesity and related metabolic syndromes via inhibition of lipid accumulation. Copyright © 2012 John Wiley & Sons, Ltd.     

基于网络药理学的人参、黄连“药对”治疗2型糖尿病作用机制研究

目的筛选人参、黄连药对主要活性成分,预测其活性成分的作用靶点,建立药物—活性成分—作用靶点网络,探讨人参、黄连药对治疗2型糖尿病(diabetes mellitus type 2,T2DM)的作用机制。方法使用TCMID、TCMSP平台筛选人参、黄连药对化学成分,并应用STITCH及DrugBank数据库检索活性成分靶点。应用TTD、DrugBank、GAD和DisGeNET数据库,筛选T2DM相关靶点。采用Cytoscape软件构建药对活性成分—作用靶点网络,通过David数据库对靶点进行生物学功能及通路富集分析。结果筛选得到人参、黄连药对潜在活性成分46个,人参相关靶点144个,黄连相关靶点246个,收集到350个T2DM相关靶点。相匹配后,得到42个人参、黄连药对治疗T2DM潜在靶点,其中,槲皮素(quercetin)、黄连素(berberine)、山奈酚(kaempferol)、花生四烯酸(arachidonate acid)、阿魏酸(ferulic acid)、豆甾醇(Stigmasterol)等6个成分能与3个及以上的关键靶点基因相连,可能是人参、黄连中治疗2型糖尿病的重要成分。结论人参、黄连药对靶点和药物代谢、胰岛素抵抗、炎症反应相关,并且参与了AMPK信号通路、NF-κB信号通路、TNF信号通路以及胰高血糖素信号通路,这些生物过程和信号通路可能是人参、黄连药对治疗T2DM的潜在机制。     

黄连解毒汤中1个新的黄酮苷

目的:研究黄连解毒汤(由黄连、黄芩、黄柏、栀子等4味中药组成的复方)的化学成分,为其药代动力学和药效动力学研究提供物质基础.方法:采用各种柱色谱方法进行分离、纯化,NMR和MS等方法进行结构鉴定.结果:从黄连解毒汤的醋酸乙酯可溶性部分得到1个新的黄酮苷,命名为汉黄芩素-5-O-β-D葡萄糖醛酸苷甲酯(wogonin-5-O-β-D-glucuronide methyl ester,1).从正丁醇可溶性部分得到10个化合物,分别鉴定为小檗碱(2),巴马汀(3),表小檗碱(4),京尼平苷(5),药根碱(6),非洲防己碱(7),格陵兰黄连碱(8),汉黄芩苷(9),3,5-二乙酰胺吡啶(10)和京尼平-1-O-β-D-龙胆双糖苷(11).结论:化合物1为新化合物.根据已报道4味中药化学成分的研究结果,判断化合物2,3和6来源于黄柏和黄连;化合物4,7和8来源于黄连;化合物5和11来源于栀子;2个黄酮苷来源于黄芩.     

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。     

基于网络药理学研究黄连-黄柏药对治疗溃疡性结肠炎的作用机制

目的:运用网络药理学技术研究黄连-黄柏药对治疗溃疡性结肠炎的主要活性成分,预测活性成分的作用靶点、建立"药对-活性成分-疾病-作用靶点"网络,探讨黄连-黄柏药对治疗溃疡性结肠炎的潜在作用机制.方法:运用中药系统药理学数据库与分析平台(TCMSP)筛选黄连-黄柏药对的化学成分,借助人类基因数据库获取溃疡性结肠炎的相关靶点...     

黄柏和附子及其单体成分对CYP3A活性的影响

目的考察黄柏、附子及其单体成分小檗碱、中乌头碱、次乌头碱对肝代谢酶CYP3A活性的影响。方法利用氯化钙沉淀法制备大鼠肝微粒体,采用红霉素脱甲基法检测大鼠CYP3A活性的变化。结果附子、黄柏的粗提物在低浓度时对CYP3A有不同程度的诱导作用,附子、黄柏在浓度为0.25,0.50 mg/ml时,其诱导作用有显著性差异;附子单体成分次乌头碱在2.5μg/ml时诱导作用最强,在25μg/ml又呈现一定的抑制作用;中乌头碱对CYP3A的诱导作用随浓度增加而增强;小檗碱作用比较缓和。结论药物粗提物及相应单体对CYP3A活性的作用趋势基本上是相同的,一定浓度黄柏和附子能增强CYP3A活性。     

消炎口服液药理作用的实验研究

消炎口服液由黄连、黄芩、黄柏、金银花、山豆根、知母、石膏等十多种中药组成。具有清热、解毒、消炎作用。药理实验表明:消炎口服液能明显抑制由角叉菜胶所致大鼠急性足趾肿胀、腹膜炎症的白细胞游出及慢性肉芽组织增生,降低伤寒菌苗致热反应。     

三黄栓质量标准研究

目的建立三黄栓的质量控制方法。方法采用薄层色谱法鉴别三黄栓中的黄连、黄芩、黄柏。采用高效液相色谱法测定制剂中黄芩苷、盐酸小檗碱的含量,色谱柱为Aglient ZORBAXSB-C18(4.6 mm×250 mm,0.45μm),甲醇-水-磷酸(47∶53∶0.2)为流动相,流速为1.0 mL/min,检测波长黄芩苷为280 nm、盐酸小檗碱为365 nm,柱温30℃。结果定性鉴别色谱特征明显。黄芩苷在0.248~2.48μg线性关系良好(r=0.999 9),平均回收率为99.77%,RSD=1.05%;盐酸小檗碱在0.336~1.68μg线性关系良好(r=0.999 9),平均回收率为97.74%,RSD=1.48%。结论该定性定量检测方法简便可行,可有效控制该制剂质量。     

梓醇和小檗碱及其配伍对脂肪细胞糖代谢和分化的影响

目的:观察中药提取物梓醇和小檗碱及其配伍对3T3-L1脂肪细胞增殖,葡萄糖代谢及细胞分化的影响,初步探讨药物改善胰岛素抵抗(IR)的可能机制。方法:采用四甲基偶氮唑盐(MTT)方法检测3T3-L1前脂肪细胞的增殖;以葡萄糖氧化酶法检测药物干预后培养液中葡萄糖消耗量,并以油红O染色检测细胞分化过程中胞浆脂质的堆积。结果:与对照组比较,小檗碱组使前脂肪细胞MTT值增加29%,而小檗碱加梓醇组、梓醇组均无差异;梓醇、小檗碱及其配伍组分别使成熟脂肪细胞的葡萄糖消耗增加64%、76.5%和98%;小檗碱处理脂肪细胞胞浆脂质堆积明显低于对照组,梓醇组与对照组无差异。结论:小檗碱能促进前脂肪细胞增殖,增加葡萄糖消耗,抑制脂肪细胞分化;梓醇能促进葡萄糖吸收;其作用均不依赖胰岛素的存在。直接增加葡萄糖的吸收可能是这两种药物降低血糖,改善胰岛素抵抗的机制之一。