减肥类中成药、保健食品、食品中非法添加酚酞、西布曲明等化学成分的快筛检测方法研究

目的建立快筛检测减肥类中成药、保健食品和食品中非法添加酚酞、西布曲明等化学成分的方法。方法用快筛试管法和薄层色谱法检测产品所含的酚酞、西布曲明等化学成分,并通过HPLC-DAD/HPLC-MS确认。结果 103批次产品中42批次检出酚酞,61批次检出西布曲明,其他相关化学成分目前均未检出。结论此方法专属性强、灵敏度高、重现性好,是一种简便快捷、经济实用、准确判断减肥类产品是否非法添加酚酞、西布曲明等化学成分的快筛检测方法,适用于快速鉴别筛查基层大批量产品。     

不同茶多糖对3T3-L1前脂肪细胞分化的抑制作用比较

本实验利用小鼠3T3-L1前脂肪细胞对绿茶多糖（green tea polysaccharides,GTP）、红茶多糖（black tea polysaccharides,BTP）和乌龙茶多糖（oolong tea polysaccharides,OTP）的减肥作用进行评价。使用气相色谱对GTP、BTP、OTP的单糖组成进行分析。采用噻唑蓝染色法测定3?种茶多糖（tea polysaccharides,TPs）对3T3-L1前脂肪细胞增殖活力的影响。使用流式细胞仪检测3?种TPs对3T3-L1前脂肪细胞细胞周期的影响。采用传统“鸡尾酒”法诱导3T3-L1细胞分化成脂后,测吸光度并计算其分化率。采用甘油磷酸氧化酶-过氧化物酶（glycerol phosphate oxidase-peroxidase,GPO-PAP）法测定细胞中甘油三酯（triglyceride,TG）含量的变化。采用实时荧光定量聚合酶链式反应（real-time polymerase chain reaction,RT-PCR）技术测定与脂质代谢相关基因的表达量。结果显示3?种TPs均能显著抑制3T3-L1前脂肪细胞的增殖与分化（P＜0.05）。添加100?μg/mL的TPs能使细胞分化率显著下降至（62.00±6.61）%（GTP）、（82.95±4.25）%（BTP）、（97.24±5.80）%（OTP）。另外,在细胞培养至第5天检测表明,这3 种TPs显著促进G0/G1期细胞数量的累积,细胞比例分别为（68.52±2.28）%（GTP）、（67.11±1.68）%（BTP）、（59.69±1.35）%（OTP）。RT-PCR分析结果显示TPs可以调控相关脂肪细胞因子的表达。在本研究中,在3T3-L1前脂肪细胞中GTP的减肥作用强于BTP和OTP。TPs的加入上调脂联素的表达从而激活腺苷酸活化蛋白激酶信号通路调控相关脂肪细胞因子的表达,并最终抑制TG的合成与3T3-L1前脂肪细胞的分化。     

Screening of crude plant extracts with anti-obesity activity

Obesity is a global health problem. It is also known to be a risk factor for the development of metabolic disorders, type 2 diabetes, systemic hypertension, cardiovascular disease, dyslipidemia, and atherosclerosis. In this study, we screened crude extracts from 400 plants to test their anti-obesity activity using porcine pancreatic lipase assay (PPL; triacylglycerol lipase, EC 3.1.1.3) in vitro activity. Among the 400 plants species examined, 44 extracts from plants, showed high anti-lipase activity using 2,4-dinitrophenylbutyrate as a substrate in porcine pancreatic lipase assay. Furthermore, 44 plant extracts were investigated for their inhibition of lipid accumulation in 3T3-L1 cells. Among these 44 extracts examined, crude extracts from 4 natural plant species were active. Salicis Radicis Cortex had the highest fat inhibitory activity, whereas Rubi Fructus, Corni Fructus, and Geranium nepalense exhibited fat inhibitory capacity higher than 30% at 100 μg/mL in 3T3-L1 adipocytes, suggesting anti-obesity activity. These results suggest that four potent plant extracts might be of therapeutic interest with respect to the treatment of obesity.     

Novel semicarbazide-sensitive amine oxidase inhibitor screening from anti-obesity drugs using HPLC-MS based technique

Semicarbazide-sensitive amine oxidase ( SSAO) has been considered to be associated with the metabolism of glucose and fat,and elevated SSAO activity was observed in obese patients. In the present study,an in vitro SSAO activity-based method was developed to screen inhibitors from 1 5 anti-obesity drugs. Among the fifteen anti-obesity drugs,four drugs including caffeine,fenfluramine, bumetanide and amfebutamone inhibited SSAO activity,and caffeine was the most effective one. When the concentration of caffeine was 1 . 4 mmol / L,the inhibition ratio was 3 1 . 9 % and 1 8 . 8 % in rabbit serum and rat adipose tissue,respectively. Inhibition of SSAO activity by caffeine was also confirmed in the in vivo study,showing the inhibition ratio of 1 5 . 6 % on serum SSAO. Caffeine provides a natural source of inhibition of SSAO activity and may be a promising inhibitor for the study of SSAO.     

茶多酚调节肠道菌群及抗肥胖作用研究进展

茶是世界上最受欢迎的饮料之一,其含有丰富的茶多酚,近来,茶多酚对肠道菌群的调节作用引起了学术界的关注。肥胖可以导致心血管疾病、糖尿病、癌症等慢性疾病,因此抗肥胖物质的研究与开发也是当下的研究热点之一,茶多酚的抗肥胖作用也是其中的重要领域。本综述总结了茶多酚、肠道菌群以及肥胖三者的作用关系,并对近5年相关的非临床以及临床研究进行了总结分析。总结发现茶多酚可以调节肠道菌群组成,而肠道菌群也可以影响茶多酚的吸收与代谢。茶多酚可以通过增加益生菌的丰度以及调节能量消耗、营养物质消化吸收、脂质分解代谢和抑制脂肪细胞合成等方式发挥抗肥胖作用。目前对于茶多酚调节肠道菌群和抗肥胖作用的研究以动物模型为主,且大多实验结果证实了茶多酚的肠道菌群调节作用及抗肥胖作用,但是作用剂量尚不明确,且茶多酚剂量毒性研究不足。另外该领域临床研究不足,需要在未来进行更大样本量和更长时间的研究,以实现茶渣等副产物的高价值利用并减少肥胖相关疾病。     

复合减肥茶及其减肥机理的研究

研究了乌龙茶以及山楂、荷叶等常见食用中药对大鼠离体激素敏感性脂肪酶和胰脂酶活性的影响, 据此确定复合减肥茶的配方,并通过动物试验检验了其减肥功效。试验结果表明, 该减肥茶能极显著地提高激素敏感性脂肪酶活性(p<0.01),增加离体脂肪释放FFA的百分率达112.9%;能有效地抑制营养肥胖型大鼠体重的增长和减少其体内脂肪含量(p<0.01);降血脂作用显著(p<0.05),减肥效果良好。     

Optimization of the Preparation of Fish Protein Anti-Obesity Hydrolysates Using Response Surface Methodology

The enzymatic condition for producing the anti-obesity hydrolysates from fish water-soluble protein was optimized with the aid of response surface methodology, which also derived a statistical model for experimental validation. Compared with neutral protease, papain and protamex, the porcine pancreas lipase inhibitory rate of hydrolysates from fish water-soluble protein was higher with alkaline protease. Results showed that the model terms were significant, the terms of lack of fit were not significant, and the optimal conditions for the hydrolysis by alkaline protease were initial pH 11, temperature 39 °C, enzyme dosage 122 U/mL and 10 h of hydrolysis time. Under these conditions, the porcine pancreas lipase and the α-amylase inhibitory rate could reach 53.04% ± 1.32% and 20.03 ± 0.89%, while predicted value were 54.63% ± 1.75%, 21.22% ± 0.70%, respectively. In addition, Lineweaver-Burk plots showed noncompetitive inhibition. The K_i value calculated was 84.13 mg/mL. These results demonstrated that fish water-soluble protein could be used for obtaining anti-obesity hydrolysates.