EGCG通过PI3K-AKT信号通路调控FOXO1磷酸化抑制3T3-L1前脂肪细胞分化

为探究EGCG对3T3-L1前脂肪细胞分化的影响及其分子机制,以3T3-L1前脂肪细胞为材料,分别添加不同浓度EGCG(0μM、5μM、10μM、50μM、100μM),通过油红O染色检测EGCG对3T3-L1脂肪细胞分化过程中脂质积累的影响,同时采用RT-PCR和免疫印迹检测不同浓度EGCG对PPARγ、FAS、PI3K、FOXO1的mRNA及蛋白表达的影响.油红O染色结果表明,EGCG可抑制3T3-L1前脂肪细胞分化,阻碍脂质积累,并具有浓度效应,EGCG浓度越高抑制作用越明显;EGCG能够直接抑制PPARγ和FAS的表达,同时抑制P-AKT、P-PI3K以及P-FOXO1的表达.表明EGCG通过介导PI3K/AKT-FOXO1级联信号通路,下调具有促进前脂肪细胞分化功能的PPARγ和FAS的表达,从而抑制前脂肪细胞分化,阻碍脂肪生成.     

普洱茶中没食子酸及其改善饮食诱导的糖脂代谢紊乱研究进展

糖脂代谢紊乱是引发心血管疾病、糖尿病和脂肪肝的重要原因之一。普洱茶中的没食子酸能够通过调节能量代谢和脂肪细胞分化、促进葡萄糖吸收与利用、提高胰岛素敏感性,进而改善饮食诱导引起的葡萄糖和脂质代谢紊乱。没食子酸通过AMPK途径、IR-Akt途径以及PPAR-γ受体调节线粒体能量代谢、胰岛素敏感性和葡萄糖吸收,从而维持糖脂代谢稳态。本文综述了普洱茶中的没食子酸及其改善饮食诱导的糖脂代谢紊乱作用和作用机制的研究进展。     

陈年六堡茶对高脂血症小鼠的调脂护肝作用研究

为了探讨陈年六堡茶的调脂保肝功效,比较分析了不同年代六堡茶的主要化学物质的差异,再以高脂高糖饮食建立高脂血症小鼠模型,并用选定的陈年六堡茶水提物灌胃小鼠,观察小鼠相关生理生化指标及组织形态的变化。结果表明,陈化15年茶样被选为陈年六堡茶的代表,用于后续试验。与模型组相比,陈年六堡茶尤其中高剂量条件下能有效减轻高脂血症小鼠体重、肝重和降低肝系数,显著提高高密度脂蛋白-胆固醇(HDL-C)含量,降低血清中总胆固醇(TC)、甘油三酯(TG)和低密度脂蛋白-胆固醇(LDL-C)含量,降低谷丙转氨酶(ALT)和谷草转氨酶(AST)活力,改善大鼠肝脏氧化应激状态,具有统计学意义(P0.05或P0.01)。肝脏和脂肪组织病理切片显示,陈年六堡茶能明显减少肝细胞中脂滴的形成和抑制脂肪细胞膨大,呈剂量依赖关系。综上表明,陈年六堡茶在正常剂量条件下能有效改善因高脂高糖饮食所致的小鼠脂质代谢紊乱和肝脏损伤。     

巴西蕉诱导的香蕉枯萎病菌1号和4号小种致病相关基因表达分析

目前对于香蕉枯萎病菌的致病分子机制尚不十分清楚。为了阐明枯萎病菌的致病分子机制,从香蕉枯萎病菌1号(Foc1)和4号小种(Foc4)的比较蛋白质组学分析发现的表达量差异较大的蛋白中,选取了9个致病相关蛋白或潜在的致病基因,利用荧光定量PCR方法进行基因表达谱分析。结果显示：与对照相比,巴西蕉处理的Foc4中,上调表达的基因有：酰胺转移酶基因（amidotransferase）、线粒体过氧化物还原酶基因（mitochondrial peroxiredoxin）、几丁质酶基因（chitinnase 1）、羧肽酶基因（carboxypeptidase cpds）,差异表达不明显的有腺苷激酶基因（adenosine kinase）、NADP-依赖型甘油脱氢酶基因（NADP-dependent glycerol dehydrogenase）、磷酸甘油酸激酶基因（phosphoglycerate kinase）、谷胱甘肽还原酶基因（glutathione reductase）、酰胺酶基因（amidase family protein）。Foc1中,与对照相比,上调表达的基因有：腺苷激酶基因、NADP-依赖型甘油脱氢酶基因,下调表达的基因有：酰胺转移酶基因、羧肽酶基因。综合分析显示,酰胺转移酶、过氧化物酶、几丁质酶、羧肽酶基因可能在Foc4的致病作用中起作用。     

大厂茶紫芽品系P113不同季节花青素调控相关基因表达分析

为明确大厂茶紫芽品系P113在不同季节花青素积累的分子机理及合成途径上相关基因的表达特点,对春、夏、秋3个季节的P113一芽二叶进行转录组（RNA-Seq）和代谢组（UHPLC-MS/MS）分析。结果显示,检测到的10种花青苷衍生物含量随季节变化,其中4种随春、夏、秋季节变化呈上调表达,与其叶色表现一致;结构基因PAL、C4H、CHS、F3'5'H表达模式基本一致,均在夏季上调表达,在秋季的表达量与夏季无显著差异;4CL酶基因获得的2个差异表达基因均在夏季上调表达;F3H、DFR、ANS表达模式相似,均在夏季上调表达,且各有1个基因在秋季呈现下调表达;2个F3'H基因中有1个在夏季下调表达,1个随季节变化上调表达;FLS的13个基因均在夏季上调表达,而在秋季呈现不同表达模式;修饰基因LAR在夏季和秋季均出现两种表达模式;ANR仅获得1个差异表达基因,随季节变化上调表达;3个UGT79B1基因中有1个在夏季下调表达,在秋季上调表达,另2个仅在夏季上调表达,在秋季的表达量与夏季无显著差异;获得的7个CCoAOMT差异表达基因均在夏季上调表达,其中2个在秋季下调表达;调控基因bHLH、MYB随季节变化上调表达,对花青素合成有正向调控作用。研究表明,大厂茶紫芽品系P113在不同季节结构基因、修饰基因和调控基因的表达具有一定的时间特性,导致了花青素积累差异。     

普洱茶功能成分单体降糖降脂作用研究

本文选取与糖脂代谢相关的PPARδ、PPARα、PPARγ、FXR、LXR、3T3-L1和α-淀粉酶模型,探讨了普洱茶中单体功能成分尿嘧啶、没食子酸的降糖降脂作用。结果表明:尿嘧啶、没食子酸在PPARγ、FXR、LXR模型表现出明显的活性作用,其中尤以没食子酸对PPARγ的激活效果最为显著,其值高达2.438,与阳性对照药物的激动效果相当,而对PPARδ、PPARα、3T3-L1模型的活性作用较弱。在PPARγ模型中,没食子酸对PPARγ受体的激活作用强于尿嘧啶,而在FXR、LXR模型中尿嘧啶的活性作用要强于没食子酸。此外,没食子酸对α-淀粉酶也有较强的抑制作用。本研究结果可为普洱茶的降糖降脂作用机理提供一定的理论依据。     

六堡茶急性和亚急性毒性安全性评价研究

六堡茶产制历史悠久,是广西特色名优农产品,但其安全性研究不够。为了探明六堡茶饮用的安全性,本实验以广西梧州六堡茶为原料,采用国家标准方法分析六堡茶常规理化成分;并按照食品安全性毒理学评价程序,进行较系统的六堡茶急性毒性和亚急性毒性实验。结果表明:六堡茶理化指标符合广西地方标准,且3个批次有较好的重复性;在小鼠急性毒性试验中,采用bliss法计算得出六堡茶半数致死剂量(LD50)为9.38 g·kg~(-1),95%的可信限为8.43~10.44 g·kg~(-1),大于5 g·kg~(-1)的经口急性毒性标准;采用大鼠30 d喂养实验发现,期间动物的生长发育良好,六堡茶3个剂量组(0.47 g·kg~(-1)·d~(-1)、0.94 g·kg~(-1)·d-1、1.88 g·kg~(-1)·d~(-1)的动物体质量、血常规指标、血生化指标、脏器指数与对照组比较,均无显著性差异(P0.05)。因此从食品毒理学的标准来看,六堡茶属于无毒级食品,符合食品安全性要求。     

水稻叶片特异性不表达启动子pOsMTG3的克隆与表达分析

利用水稻基因芯片筛选到1个在水稻孕穗期和抽穗期穗中的相对表达量明显高于苗期、孕穗期和抽穗期叶片中相对表达量的基因Os MTG3,该基因在孕穗期穗中受低温、干旱诱导上调表达,而在抽穗期穗中受低温诱导下调表达。把该基因ATG密码子上游1.8kb左右的DNA片段预测为启动子区,并将其命名为p Os MTG3。用PCR技术克隆该启动子,并以GUS基因作为报告基因,在水稻中分析了该启动子在不同时期不同组织中的表达特性。GUS组织化学染色分析表明:p Os MTG3启动子能启动GUS基因在水稻根、芽、茎、穗中表达,而不能启动GUS基因在叶片中表达,是一个全新的叶片特异性不表达的启动子。     

利用Affymetrix芯片分析DON诱导抗赤霉病小麦品种望水白的基因表达谱

中国小麦地方品种望水白不但高抗赤霉病,而且其籽粒中DON(脱氧雪腐镰刀菌烯醇,被认为是赤霉病的致病因子)含量也低.为了研究望水白低DON含量的分子机制,利用Affymetrix小麦基因芯片对望水白受DON诱导调控的基因进行高通量的检测,以DON诱导后12 h和24 h混合样品作为处理组,水处理做为对照.总共检测到差异表达的基因1 114个,其中上调表达的基因有949个,下调基因165个.在上调表达基因中,推断有功能的基因涉及转录因子、信号蛋白、着丝粒蛋白以及与病程相关的基因,如:腺苷三磷酸结合盒转运蛋白,谷胱甘酞转移酶、细胞色素P450酶、苯丙氨酸解氨酶、葡萄糖基转移酶以及抗病蛋白等.对上调表达中的部分基因进行RT-PCR分析,证实它们都受DON诱导上调表达,与芯片检测结果吻合.     

化学杀雄剂CH1诱导的小麦雄性不育花药转录组学及相关基因表达的分析

为了揭示化学杀雄剂CH1诱导雄性不育的作用机理,以小麦品种普冰151为试验材料,利用测序技术对化杀不育系(FHS)和正常可育系(FZC,对照)的花药RNA进行转录组学分析,筛选差异表达基因,并进行GO、KEGG富集分析及实时荧光定量PCR分析。结果表明,FHS和FZC之间筛选出差异表达基因共3 895个,其中上调表达基因1 334个,下调表达基因2 561个,主要富集在碳水化合物代谢、多糖代谢、外部结构组织、细胞壁组织等;经KEGG分析,差异基因主要集中在戊糖和葡萄糖醛酸转化、淀粉和蔗糖代谢利用等通路中。利用实时定量PCR技术对转录组数据中所筛选出的5个表达差异较大的基因进行验证,在花药发育的单核期,处理与对照相比,这5个基因表现出不同的上下调趋势;而在二核期和三核期,这5个基因都表现出明显的下调趋势,推测这5个基因的下调表达有可能与化杀不育系花粉的败育有关。     

Ameliorating Effects of Fermented Rice Bran Extract on Oxidative Stress Induced by High Glucose and Hydrogen Peroxide in 3T3-L1 Adipocytes

In this study, we investigated whether fermented rice bran (FRB) can ameliorate the oxidative stress induced by high glucose and hydrogen peroxide (H₂O₂) in 3T3-L1 adipocytes by analyzing reactive oxygen species (ROS), oil red O staining, as well as the expression of mRNAs related to glucose homeostasis and adipogenesis. It was first confirmed that rice bran fermented by Issatchenkia orientalis MFST1 extract increased free phenolic content compared to non-fermented rice bran. The FRB extract strongly inhibited ROS generation and upregulated the expression of PPAR-γ and adiponectin. Moreover, FRB upregulated GLUT4 related to glucose transportation and insulin sensitivity. Taken together, FRB extract ameliorated oxidative stress-induced insulin resistance by neutralizing free radicals and upregulating adiponectin in adipocytes. Our results provide information toward understanding the beneficial effects of FRB on oxidative stress.     

Shrimp Oil Extracted from Shrimp Processing By-Product Is a Rich Source of Omega-3 Fatty Acids and Astaxanthin-Esters,and Reveals Potential Anti-Adipogenic Effects in 3T3-L1 Adipocytes

The province of Newfoundland and Labrador, Canada, generates tons of shrimp processing by-product every year. Shrimp contains omega (n)-3 polyunsaturated fatty acids (PUFA) and astaxanthin (Astx), a potent antioxidant that exists in either free or esterified form (Astx-E). In this study, shrimp oil (SO) was extracted from the shrimp processing by-product using the Soxhlet method (hexane:acetone 2:3). The extracted SO was rich in phospholipids, n-3 PUFA, and Astx-E. The 3T3-L1 preadipocytes were differentiated to mature adipocytes in the presence or absence of various treatments for 8 days. The effects of SO were then investigated on fat accumulation, and the mRNA expression of genes involved in adipogenesis and lipogenesis in 3T3-L1 cells. The effects of fish oil (FO), in combination with Astx-E, on fat accumulation, and the mRNA expression of genes involved in adipogenesis and lipogenesis were also investigated. The SO decreased fat accumulation, compared to untreated cells, which coincided with lower mRNA expression of adipogenic and lipogenic genes. However, FO and FO + Astx-E increased fat accumulation, along with increased mRNA expression of adipogenic and lipogenic genes, and glucose transporter type 4 (Glut-4), compared to untreated cells. These findings have demonstrated that the SO is a rich source of n-3 PUFA and Astx-E, and has the potential to elicit anti-adipogenic effects. Moreover, the SO and FO appear to regulate adipogenesis and lipogenesis via independent pathways in 3T3-L1 cells.     

Lipid Inhibitory Effect of (−)-loliolide Isolated from Sargassum horneri in 3T3-L1 Adipocytes: Inhibitory Mechanism of Adipose-Specific Proteins

Sargassum horneri (S. horneri) is a well-known brown seaweed widely distributed worldwide. Several biological activities of S. horneri have been reported. However, its effects on lipid metabolism and the underlying mechanisms remain elusive. In the present study, we examined the inhibitory effect of the active compound “(−)-loliolide ((6S,7aR)-6-hydroxy-4,4,7a-trimethyl-5,6,7,7a-tetrahydro-1-benzofuran-2(4H)-one (HTT))” from S. horneri extract on lipid accumulation in differentiated adipocytes. MTT assays demonstrated that (−)-loliolide is not toxic to 3T3-L1 adipocytes in a range of concentrations. (−)-loliolide significantly reduced intracellular lipid accumulation in the differentiated phase of 3T3-L1 adipocytes as shown by Oil Red O staining. Western blot analysis revealed that (−)-loliolide increased the expression of lipolytic protein phospho-hormone-sensitive lipase (p-HSL) and thermogenic protein peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1). Additionally, (−)-loliolide decreased expression of adipogenic and lipogenic proteins, including sterol regulatory element-binding protein-1 (SREBP-1), peroxisome proliferator-activated receptor-γ (PPAR-γ), CCAAT/enhancer-binding protein-α (C/EBP-α), and fatty acid-binding protein 4 (FABP4) in 3T3-L1 adipocytes. These results indicate that (−)-loliolide from S. horneri could suppress lipid accumulation via regulation of antiadipogenic and prolipolytic mechanisms in 3T3-L1 cells. Considering the multifunctional effect of (−)-loliolide, it can be useful as a lipid-lowering agent in the management of patients who suffer from obesity.     

六堡茶对高脂饮食大鼠肠道短链脂肪酸含量的影响

探讨六堡茶对高血脂症模型大鼠肠道短链脂肪酸的影响。以低剂量50 mg·kg^-1·d^-1、中剂量100 mg·kg^-1·d^-1和高剂量200 mg·kg^-1·d^-1六堡茶水提物对高脂模型大鼠进行干预性治疗4周后,收集大鼠粪便;将六堡茶醇提物与高脂模型空白组大鼠粪便进行体外发酵培养,最后采用气相色谱法分别测定粪便和发酵液短链脂肪酸含量。体内试验结果表明,六堡茶水提物能显著降低高脂大鼠肠道的乙酸含量,显著增加高脂大鼠肠道的丙酸和丁酸的含量;与绿茶水提物相比,六堡茶对高脂大鼠短链脂肪酸的影响更强。体外试验结果表明,六堡茶醇提物对乙酸的抑制率为16.92%,对丙酸和丁酸的促进率分别为24.81%和23.03%;4种茶叶中,六堡茶对肠道短链脂肪酸的影响最大,普洱熟茶次之,六堡原料毛茶和绿茶的影响最小;六堡茶醇提物不同极性段组分均能抑制乙酸分泌、促进丙酸和丁酸的分泌,其中,六堡茶醇提物通过60%的乙醇洗脱组分对肠道短链脂肪酸的影响最大。     

茯茶水提物对Ⅱ型糖尿病小鼠糖代谢紊乱的干预作用

采用高脂高糖饲料联合链脲佐菌素诱导Ⅱ型糖尿病小鼠模型,研究茯砖茶水提物对Ⅱ型糖尿病小鼠的降糖功效。结果表明,茯茶水提取物干预能显著改善糖尿病小鼠多食、多饮、多尿、消瘦症状,降低空腹血糖浓度,提高机体对糖负荷的耐受性,提升胰岛素和胰岛素敏感水平,降低胰岛素抵抗水平。同时,q RT-PCR分析发现,茯茶水提物通过增强PPAR-α、GLUT2和GLUT4基因表达,促进了葡萄糖转运和肝脏内脂质代谢,从而改善糖尿病小鼠体内糖代谢紊乱。综上所述,茯茶能显著改善Ⅱ型糖尿病小鼠糖代谢紊乱症状,其中中、高剂量茯茶效果最为显著。     

Fucoidan from marine brown algae inhibits lipid accumulation

In this study, we elucidated the inhibitory effect of fucoidan from marine brown algae on the lipid accumulation in differentiated 3T3-L1 adipocytes and its mechanism. The treatment of fucoidan in a dose-dependent manner was examined on lipid inhibition in 3T3-L1 cells by using Oil Red O staining. Fucoidan showed high lipid inhibition activity at 200 μg/mL concentration (P < 0.001). Lipolytic activity in adipocytes is highly dependent on hormone sensitive lipase (HSL), which is one of the most important targets of lipolytic regulation. Here, we examined the biological response of fucoidan on the protein level of lipolysis pathway. The expressed protein levels of total hormone sensitive lipase (HSL) and its activated form, phosphorylated-HSL were significantly increased at concentration of 200 μg/mL fucoidan. Furthermore, insulin-induced 2-deoxy-D-[3H] glucose uptake was decreased up to 51% in fucoidan-treated cells as compared to control. Since increase of HSL and p-HSL expression and decrease of glucose uptake into adipocytes are known to lead to stimulation of lipolysis, our results suggest that fucoidan reduces lipid accumulation by stimulating lipolysis. Therefore, these results suggest that fucoidan can be useful for the prevention or treatment of obesity due to its stimulatory lipolysis.     

茶多酚防治2型糖尿病的分子机理研究进展

茶多酚作为天然功能性物质能有效防治2型糖尿病,但是其具体作用机制仍不太清楚。研究表明它的作用机理主要是通过调控糖代谢平衡：包括降低α-糖苷酶、α-淀粉酶等双糖酶活性,调控胰岛素信号传导途径及AMPK途径中多个分子靶点的磷酸化水平,影响酶与转录因子的活性与表达,减弱胰岛素抵抗及糖异生作用,促进胰岛素的合成与分泌,促进靶组织对葡萄糖的吸收与利用等;同时,茶多酚能够调控脂代谢,通过抑制脂质合成与积累的相关酶的活性及转录因子的表达,促进脂质的氧化代谢,减轻脂代谢紊乱;另外,茶多酚的抗氧化与抗炎作用可以有效减轻组织细胞的氧化伤害和炎症损伤,减少细胞凋亡。     

茶多糖和茶多酚的降血糖作用研究

目的:研究茶多糖、茶多酚对四氧嘧啶致糖尿病SD大鼠的降血糖作用和机制。方法:饲喂SD大鼠茶多糖、茶多酚3周后,观察大鼠血糖、葡萄糖耐量、血胰岛素以及小肠糖降解酶(淀粉酶、蔗糖酶、麦芽糖酶)变化。结果:茶多糖、茶多酚都有显著抑制糖尿病大鼠血糖升高的作用;与对照组比较,茶多糖组大鼠血胰岛素水平有显著提高(P<0.05),蔗糖酶和麦芽糖酶活性显著降低(P<0.05);茶多酚组的血胰岛素水平有升高趋势,小肠各降解酶活力也有下降趋势,但与对照组比较均未达到显著水平。结论:茶多糖对高血糖大鼠有显著的抑制血糖升高的作用,茶多糖的作用机制可能是抑制小肠糖降解酶活性。