Anti-obesity effect of sulfated glucosamine by AMPK signal pathway in 3T3-L1 adipocytes

In this study, we investigated the effect of sulfated glucosamine (SGlc) on adipogenesis of 3T3-L1 adipocytes during differentiation of preadipocytes into adipocytes by measuring lipid accumulation and adipogenesis related factors. Treatment with SGlc reduced the triglyceride content in Oil-Red O staining and enhanced glycerol secretion in adipocytes in a dose-dependent manner. In addition, SGlc induced the down-regulation of adipogenesis related factors and adipocyte specific gene promoters. Moreover, treatment of 3T3-L1 adipocytes with SGlc activated the phosphorylated adenosine monophosphate-activated protein kinase (AMPK) α and β along with their substrate, acetyl-CoA carboxylase (ACC). These results suggest that inhibitory effect of SGlc on adipocyte differentiation might be mediated through the up-regulation of AMPK pathway.     

黄芩水提液对3T3-L1脂肪细胞增殖、诱导分化及脂联素启动子荧光素酶活性的影响

目的：本研究旨在观察黄芩水提液（Scutellaria Baicalensis Water Extract,SBWE）对3T3-L1前体细胞增殖、分化,对脂肪细胞因子脂联素表达以及脂联素（Adiponectin,ADP）启动子荧光素酶活性的影响,从分子生物学角度阐述SBWE降脂作用的可能机理。方法：通过体外培养3T3-L1细胞,采用MTT法检测SBWE对3T3-L1细胞增殖能力的影响;通过诱导脂肪细胞分化成为成熟脂肪细胞,观察SBWE对脂肪形成的影响;化学发光法检测脂联素启动子双荧光素酶报告基因活性;荧光定量PCR法检测脂联素mRNA（Adipoq）表达。结果: 与正常组相比,给予3T3-L1细胞0.01、0.1、1 mg?mL^-1浓度的SBWE 24 h,可显著抑制细胞的增殖活性（P<0.05）;0.1、1 mg?mL^-1浓度的SBWE能够降低3T3-L1细胞分化为脂肪细胞的数量,并减少细胞内脂滴聚集,但无明显剂量依赖性;0.01、0.1 mg?mL^-1浓度SBWE能显著提高脂联素基因启动子荧光素酶活性,与空载体比较差异有统计学意义（P<0.05）;与正常组相比,给予3T3-L1细胞0.1 mg?mL^-1 SBWE 24 h,诱导前后的脂肪细胞Adipoq表达均明显增加 （P<0.05）。结论: SBWE可有效抑制3T3-L1脂肪细胞的增殖、分化,同时增加脂联素基因表达,这可能是通过增强脂联素基因启动子荧光素酶活性实现,这些为黄芩水提液减肥的作用机制提供一定的基础。     

小檗碱对白介素-6诱导胰岛素抵抗 3T3-L1脂肪细胞脂联素表达的影响

目的:观察小檗碱对白介素-6(IL-6)诱导胰岛素抵抗的3T3-L1脂肪细胞脂联素表达的影响。方法:选用IL-6诱导3T3-L1脂肪细胞胰岛素抵抗(IR)模型。以20μg·L-1IL-6培养48 h,将3T3-L1脂肪细胞随机分为正常对照组、模型组、吡格列酮组(50μmol.L-1)和小檗碱高、中、低剂量组(10,20,50μmol.L-1),以葡萄糖氧化酶法测定葡萄糖消耗量,观察小檗碱对脂肪细胞葡萄糖摄取的影响,鉴定IR模型;采用实时荧光定量PCR技术测定脂肪细胞脂联素基因mRNA水平。结果:模型组葡萄糖消耗量及脂联素基因表达水平与正常对照组比较,均显著降低(P<0.05);小檗碱高、中、低剂量组及吡格列酮组均能显著增加葡萄糖消耗量及脂联素基因表达水平(P<0.05)。结论:小檗碱可增加白介素-6诱导胰岛素抵抗的3T3-L1脂肪细胞脂联素基因mRNA的表达,改善胰岛素抵抗状态。     

原发性肝癌病人AFP-L3、AFU、GP73及TSGF等血清标志物的水平分析及临床意义

目的 评价血清甲胎蛋白异质体(AFP-L3)、α-L-岩藻糖苷酶(AFU)、高尔基体糖蛋白73(GP73)及肿瘤特异性生长因子(TSGF)四者联合在原发性肝癌(PHC)诊断中的价值.方法 选取2015年3月至2016年3月间我院收治的90例PHC患者作为观察组,并选择同一时期收治的45例肝炎肝硬化患者作为疾病对照组,及45例健康志愿者作为健康对照组.采取微量离心法分离血清AFP-L3,然后检测其占总甲胎蛋白(AFP)的百分比,生化定量法测三组血清AFU、TSGF水平,以及酶联免疫法(ELISA)测三组血清GP73水平.结果 观察组血清AFP-L3、AFU、GP73及TSGF水平均明显高于疾病对照组与健康对照组,差异有统计学意义(P＜0.05);观察组上述四种指标单独及联合检测的阳性率均显著高于疾病对照组与健康对照组的检测阳性率,差异有统计学意义(P＜0.05);同时AFP-L3、AFU、GP73及TSGF检测的灵敏度分别为58.9％、72.2％、74.4％、64.4％,特异性分别为90.0％、75.6％、76.7％、81.1％,准确度分别为74.4％、73.9％、75.6％、72.8％,而四者联合的灵敏度、特异性及准确度分别为92.2％、96.7％、94.4％,联合检测明显高于各指标单独检测,差异有统计学意义(P＜0.05).结论 血清AFP-L3、AFU、GP73及TSGF联合检测能够显著增强检测的灵敏度及特异性,提高PHC的检出率.     

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。     

胰岛素、葡萄糖对3T3-L1脂肪细胞脂肪水孔蛋白表达的调节

利用半定量RT PCR技术及Western印迹法研究胰岛素、葡萄糖对成熟脂肪细胞脂肪水孔蛋白 (AQPap)基因表达的影响。结果表明 ,胰岛素对AQPap的表达具有抑制作用 ;而高浓度葡萄糖则对AQPap的表达具有促进作用     

Multiple molecular targets underlie the antidiabetic effect of Nigella sativa seed extract in skeletal muscle, adipocyte and liver cells

Aim: Nigella sativa ( N. sativa ) is a plant widely used in traditional medicine of North African countries. During the last decade, several studies have shown that extracts from the seeds of N. sativa have antidiabetic effects.
Methods: Our group has recently demonstrated that N. sativa seed ethanol extract (NSE) induces an important insulin-like stimulation of glucose uptake in C2C12 skeletal muscle cells and 3T3-L1 adipocytes following an 18 h treatment. The purpose of the present study was to elucidate the pathways mediating this insulin-like effect and the mechanisms through which these pathways are activated.
Results: Results from western immunoblot experiments indicate that in C2C12 cells as well as in H4IIE hepatocytes, but not in 3T3-L1 cells, NSE increases activity of Akt, a key mediator of the effects of insulin, and activity of AMP-activated protein kinase (AMPK), a master metabolic regulating enzyme. To test whether the activation of AMPK resulted from a disruption of mitochondrial function, the effects of NSE on oxygen consumption were assessed in isolated liver mitochondria. NSE was found to exhibit potent uncoupling activity.
Conclusion: Finally, to provide an explanation for the effects of NSE in adipocytes, PPARγ stimulating activity was tested using a reporter gene assay. Results indicate that NSE behaves as an agonist of PPARγ. The data supports the ethnobotanical use of N. sativa seed oil as a treatment for diabetes, and suggests potential uses of this product, or compounds derived thereof, against obesity and the metabolic syndrome.