菠萝叶酚对小鼠急性糖脂负荷吸收及其小肠脂肪酶活性的影响

目的：研究表明从菠萝叶中制备的总酚类成分具有抗氧化和调血脂作用。由于其灌胃后吸收较快,血中5 min 即达峰值,因此本文通过急性糖脂负荷实验观察菠萝叶酚对糖脂吸收的影响。方法：利用小鼠单次灌胃建立糖脂负荷模型,观察给药后不同时间对餐后血糖、血脂和小肠脂肪酶的变化。结果：菠萝叶酚在一定程度上能够抑制甘油三酯和葡萄糖的吸收。在给药30 min 后未见到明显的作用,但此时正常小鼠小肠脂肪酶活性受到明显抑制,并且对高脂所致小肠脂肪酶活性下降有一定的解抑制作用。结论：提示菠萝叶酚的抑制脂类吸收可能与脂肪酶活性有关;其对于餐后高糖高脂吸收的有一定调控作用。     

新药用资源菠萝叶的质量标准研究

目的：对菠萝叶质量标准进行研究,以更好的开发应用菠萝叶的药用资源。方法：利用组织切片法观察菠萝叶组织结构;利用薄层层析法以菠萝酰酯为对照品进行定性鉴别;利用高效液相色谱法对主要成分进行含量测定。结果：菠萝叶镜下可分上下表皮层,其间有气孔;薄层层析可以清晰检测到菠萝酰酯成分;高效液相色谱法可以准确的测定菠萝叶中对香豆酸和菠萝酰酯的含量,且不同产地的菠萝叶中对香豆酸和菠萝酰酯含量差异较大,其中以云南的菠萝叶含量最高,广西的菠萝叶含量最低。结论：以菠萝酰酯作定性鉴别,以对香豆酸和菠萝酰酯为指标成分进行定量检测,可以较好的控制菠萝叶药材的质量。     

石榴叶鞣质和鞣花酸体外对大鼠原代脂肪细胞转化及脂代谢相关因子表达的影响

目的：探讨石榴叶鞣质及其鞣花酸对原代大鼠脂肪细胞分化及其相关脂代谢影响因子表达的影响。方法：利用体外培养原代大鼠前脂肪细胞方法,观察石榴叶鞣质及其鞣花酸对脂肪细胞内脂生成及其相关因子mRNA表达的影响。结果：石榴叶鞣质和鞣花酸对脂肪细胞中脂的形成具有明显的抑制作用,对脂蛋白脂酶（LPL）和葡萄糖-3-磷酸脱氢酶（GPDH）活性有一定的抑制作用,促进脂分解,降低胞内脂含量。并且上调PPARγ和脂肪酸结合蛋白（aP2）,下调肥胖基因（ob）。结论：石榴叶鞣质可以抑制脂肪细胞内脂的生成,并能促进脂代谢,鞣花酸是其主要活性成分,具有与石榴叶鞣质相同的作用。     

石榴叶鞣质对高血脂小鼠糖代谢影响及其机制

目的：探讨石榴叶鞣质对高脂小鼠糖代谢的影响以及降糖的作用机制。方法：通过高脂饮食建立高脂并高血糖小鼠,灌胃石榴叶鞣质,观察小鼠血清糖脂的变化,肝糖元含量;通过体外HepG2细胞实验观察石榴叶鞣质对葡萄糖的利用及其胞内糖元含量变化。结果：灌胃石榴叶鞣质8周后,高脂小鼠外周血总胆固醇、甘油三酯和血糖均有明显降低,肝糖元升高;体外实验石榴叶鞣质可以明显促进HepG2细胞对葡萄糖的利用,增加胞内糖元含量。结论：石榴叶鞣质对高脂合并的高血糖有明显的降低作用,其作用机制可能与增加肝内糖元转化有关。     

石榴叶鞣质对高血脂小鼠糖代谢影响及其机制

目的：探讨石榴叶鞣质对高脂小鼠糖代谢的影响以及降糖的作用机制。方法：通过高脂饮食建立高脂并高血糖小鼠,灌胃石榴叶鞣质,观察小鼠血清糖脂的变化,肝糖元含量;通过体外HepG2细胞实验观察石榴叶鞣质对葡萄糖的利用及其胞内糖元含量变化。结果：灌胃石榴叶鞣质8周后,高脂小鼠外周血总胆固醇、甘油三酯和血糖均有明显降低,肝糖元升高;体外实验石榴叶鞣质可以明显促进HepG2细胞对葡萄糖的利用,增加胞内糖元含量。结论：石榴叶鞣质对高脂合并的高血糖有明显的降低作用,其作用机制可能与增加肝内糖元转化有关。     

石斛酚在人晶状体上皮细胞中跨膜吸收机制研究

石斛酚为中药石斛的酚类成分，具有抗肿瘤、抗糖尿病性白内障等多种药理活性。该文探索其在人晶状体上皮细胞(HLECs)中跨膜转运的分子机制。体外培养永生化的HLECs,按5 000个/mL细胞接种于激光共聚焦扫描显微镜(LSCM)培养皿中，通过LSCM观察荧光标记的石斛酚在HLECs内的荧光分布和荧光强度，以荧光强度表示石斛酚的吸收量，监测石斛酚在HLECs内的跨膜转运过程，比较时间、温度、浓度、转运抑制剂以及不同细胞系对石斛酚跨膜吸收和转运的影响；HLECs接种于6孔培养板的爬片上，原子力显微镜(AFM)检测非荧光标记的石斛酚跨膜吸收过程中HLECs表面超微结构的变化。结果显示，石斛酚跨膜吸收存在时间、温度、浓度依赖性，并能够专一性地靶向识别HLECs;能量及载体转运抑制剂降低HLECs对石斛酚的吸收量；在石斛酚跨膜过程中，HLECs细胞膜表面变得更粗糙，并呈现不同程度的凹坑。结果表明，石斛酚跨膜转运方式是通过能量及载体介导的胞吞作用主动吸收实现的。     

急弯棘豆化学成分的研究

从急弯棘豆Oxytropisdeflexa（Pall）DC．全草中分离得到9个化合物，经理化常数和光谱分析确定其结构为：蜂花酸（Ⅰ），β-谷甾醇（Ⅱ），五味酯甲（Ⅲ），山奈酚（Ⅳ），槲皮素（Ⅴ），山奈酚-3-O－6″-乙酸吡喃葡萄糖甙（Ⅵ），紫云英式（Ⅶ），异槲皮甙（Ⅷ），山奈酚-3-O-6″-丙二酰毗喃葡萄糖甙（Ⅸ），上述成分均首次从该植物中分得，其中木脂素Ⅲ和黄酮甙Ⅵ，Ⅸ在豆科棘豆属植物中首次发现。     

三七根际土壤中酚酸类物质的鉴定及含量测定

目的：对云南文山三七根际土壤中酚酸的种类进行鉴定,并对其进行含量测定。方法：采用HPLC-MS、HPLC 技术,并通过文献和对照品比对,鉴定了云南文山三七根际土壤中酚酸的种类并对其进行含量测定。结果：从三七根际土壤提取液中检测到对羟基苯甲酸、香草酸、丁香酸、对香豆酸、阿魏酸和苯甲酸6 种酚酸类物质,其中对香豆酸含量最高（24.01 μg·g-1）,丁香酸含量最低（1.26 μg·g-1）。6 种酚酸类成分在三七根际土壤中的含量顺序为对香豆酸>香草酸>苯甲酸>对羟基苯甲酸>阿魏酸>丁香酸。结论：三七根际土壤中的酚酸类物质可能为三七植物须根残体腐解产生,也可能是三七根系分泌产生。     

肝细胞亲和色谱法体外筛选大黄降脂活性成分研究

目的：评价大黄蒽醌类成分体外降脂活性及筛选其药效成分,探讨其在细胞内的成分代谢。方法：采用体外HepG2细胞系脂肪变性模型评价大黄蒽醌类成分降脂活性,采用肝细胞亲和色谱法筛选大黄蒽醌类细胞亲和成分,以HPLC法及HPLC-Q-TOF-MS法检测鉴定亲和成分。结果：与正常组相比,建立的体外肝细胞脂肪变性模型组细胞内三酰甘油（TG）含量显著升高（P<0.05）,与模型组比较,不同浓度的五种大黄蒽醌类成分作用后给药组细胞内TG含量均显著降低（P<0.05）,呈剂量效应关系;肝细胞亲和色谱法筛选出大黄提取液特异性亲和色谱成分主要有4个,分别为芦荟大黄素、大黄素、大黄酚、大黄素甲醚四个化合物,而大黄酸发生了细胞内代谢。结论：大黄蒽醌类5种成分均具有体外降脂活性,芦荟大黄素、大黄素、大黄酚、大黄素甲醚以原型的形式体外肝细胞降脂,而大黄酸可能以细胞内代谢为芦荟大黄素形式降脂。本研究建立了肝细胞亲和色谱法与体外肝细胞降脂活性评价的联用技术,实现了中药成分体外降脂活性评价,药效成分体外代谢的一体化研究,为大黄蒽醌类调脂药物开发提供科学依据。     

中药防治实验性肝纤维化研究概况

近年来，中医药抗肝纤维化研究较多，本文对实验性肝纤维化有防治作用的单味药、复方及其作用予以简介。单味药及其提取物汉防已甲素是从防已种植物根中提取的一种生物碱，主要作用于钙离子通道，影响钙离子的跨膜转运以及在细胞内的分布利用。上海第二医科大学附属新华医院消化内科[1～2]，观察到它可降低CCL4肝纤维化大鼠肝羟脯氨酸、透明质酸含量和N-乙酸-β氨基葡萄糖苷酶活性，预防应用效果更佳，而且可使纤维化肝脏中的胶原纤维、网状纤维的平均面积、肝储脂细胞、枯否氏细胞及其粗面内质网的平均面积减少，揭示汉防已甲素（Tet）的…     

SDF7, a group of Scoparia dulcis Linn. derived flavonoid compounds,stimulates glucose uptake and regulates adipocytokines in 3T3-F442a adipocytes

Ethnopharmacology relevance

Adipocytes are major tissues involved in glucose uptake second to skeletal muscle and act as the main adipocytokines mediator that regulates glucose uptake mechanism and cellular differentiation. The objective of this study were to examine the effect of the SDF7, which is a fraction consists of four flavonoid compounds (quercetin: p-coumaric acid: luteolin: apigenin=8: 26: 1: 3) from Scoparia dulcis Linn., on stimulating the downstream components of insulin signalling and the adipocytokines expression on different cellular fractions of 3T3-F442a adipocytes.

Material and methods

Morphology and lipid accumulation of differentiated 3T3-F442a adipocytes by 100 nM insulin treated with different concentrations of SDF7 and rosiglitazone were examined followed by the evaluation of glucose uptake activity expressions of insulin signalling downstream components (IRS-1, PI3-kinase, PKB, PKC, TC10 and GLUT4) from four cellular fractions (plasma membrane, cytosol, high density microsome and low density microsome). Next, the expression level of adipocytokines (TNF-α, adiponectin and leptin) and immunoblotting of treated 3T3-F442 adipocytes was determined at 30 min and 480 min. Glucose transporter 4 (GLUT4) translocation of 3T3-F442a adipocytes membrane was also determined. Lastly, mRNA expression of adiponectin and PPAR-γ of 3T3-F442a adipocytes were induced and compared with basal concentration.

Results

It was found that SDF7 was able to induce adipocytes differentiation with great extends of morphological changes, lipid synthesis and lipid stimulation in vitro. SDF7 stimulation of glucose transport on 3T3-F442a adipocytes are found to be dose independent, time-dependent and plasma membrane GLUT4 expression-dependent. Moreover, SDF7 are observed to be able to suppress TNF-α and leptin expressions that were mediated by 3T3-F442a adipocytes, while stimulated adiponectin secretion on the cells. There was a significant expression (p<0.01) of protein kinase C and small G protein TC10 on 3T3-F442a adipocytes upon treatment with SDF7 as compared to the control. SDF7 was also found to be effective in stimulating adiponectin and PPAR-γ mRNA upregulation at 50 µg/ml.

Conclusion

SDF7 exhibited good lipogenesis, adiponectinesis and glucose uptake stimulatory properties on 3T3-F442a adipocytes.     

Regulation effects on abnormal glucose and lipid metabolism of TZQ-F,a new kind of Traditional Chinese Medicine

Aim of the study

Based on the recipe of the traditional anti-diabetic formula TZQ, we developed TZQ-F, a new formula including 8 fractions isolated from Red Paeony root, Mulberry leaf, Lotus leaf, Danshen root and Hawthorn leaf with a good quality assurance. The study was aimed at fraction preparation and effects of the fractions on abnormal glucose and lipid metabolism.

Materials and methods

The active fractions were obtained by macroporous resin, ion-exchange resin and polyamide resin column chromatographies. HPLC analyses were used for quality control. In vitro mechanism study included DPPH radical scavenging, AGEs formation inhibition, α-glucosidase inhibition and lipase inhibition, and rats on high-fat diet were used for in vivo study.

Results

In vitro mechanism study showed that among the 8 fractions, three of them had inhibition effects on intestinal disaccharase, three with inhibition effects on lipase, and five with effects of free radical scavenging. In vivo study showed that after 4 weeks of treatment, TZQ-F significantly decreased the levels of serum total cholesterol, TG, glucose, LDL-C and HDL-C in rats on high-fat diet. Consistent with the in vitro and in vivo results, histology study demonstrated that TZQ-F alleviated hepatic steatosis induced by high-fat diet.

Conclusions

TZQ-F possesses the potential regulation effects on abnormal glucose and lipid metabolism.     

黑果枸杞花色苷对小鼠动脉粥样硬化的影响

目的:研究黑果枸杞花色苷对小鼠动脉粥样硬化(AS)的影响。方法:以正常小鼠作空白对照,将高脂血症模型小鼠分为高脂模型组、黑果枸杞花色苷低、中、高剂量组和辛伐他汀药物对照组,进行不同灌胃处理,酶法检测各组小鼠血脂指标;苏木素伊红(HE)染色观察主动脉、心脏及肝脏组织形态学变化,并对肝脏相关指标进行测定。结果:与高脂组对照,花色苷各剂量组小鼠血清的总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)和动脉粥样硬化指数(AI)水平均显著降低(P<0.05),高密度脂蛋白胆固醇(HDL-C)水平有高于模型组的趋势。肝脏的总抗氧化能力(T-AOC),谷胱甘肽过氧化物酶(GSH-PX)、脂蛋白酯酶(LPL)均显著升高(P<0.05),丙二醛(MDA)极显著下降(P<0.01);花色苷各剂量组主动脉斑块面积占总面积的百分比显著低于模型组(P<0.05);主动脉、心脏和肝脏的病变程度明显轻于高脂组,其中,中剂量组与辛伐他汀组类似。结论:黑果枸杞花色苷可以干预小鼠AS的形成,同时降低小鼠血脂水平。     

氮、磷、钾缺乏对西洋参根分泌物中 酚酸类化合物的影响

目的:研究氮、磷、钾元素缺乏对西洋参根系分泌物中具有自毒作用的酚酸类化合物的影响.方法:以二年生健康西洋参幼苗为试验材料,设计Hoagland全营养液以及分别去除氮、磷、钾元素4个处理进行水培试验,2周后收集不同处理的培养液,用XAD4和XAD7离子交换树脂富集其中不同极性的根系分泌物.Folin-Ciocalteu试剂法测定不同根系分泌物中总酚酸含量,HPLC测定香草酸、对香豆酸、肉桂酸3种单体酚酸含量.结果:与全营养条件相比,缺氮或缺钾情况下西洋参根分泌物中总酚酸含量显著升高(P<0.05);而磷的缺乏可导致西洋参根分泌物中总酚酸含量显著降低(P<0.05);缺乏3种元素之中的任何一种均可使香草酸、对香豆酸、肉桂酸在根系分泌物中的比例显著降低(P<0.05).结论:氮、磷、钾的缺乏可导致西洋参根系分泌物中的酚酸发生不同变化.     

Antitubercular activity of Arctium lappa and Tussilago farfara extracts and constituents

Ethnopharmacological relevance

Arctium lappa and Tussilago farfara (Asteraceae) are two plant species used traditionally as antitubercular remedies. The aim of this study was (i) to screen Arctium lappa and Tussilago farfara extracts for activity against Mycobacterium tuberculosis and (ii) to isolate and identify the compound(s) responsible for this reputed anti-TB effect.

Materials and methods

The activity of extracts and isolated compounds was determined against Mycobacterium tuberculosis H₃₇R_v using a high throughput spot culture growth inhibition (HT-SPOTi) assay.

Results

The n-hexane extracts of both plants, the ethyl acetate extract of Tussilago farfara and the dichloromethane phase derived from the methanol extract of Arctium lappa displayed antitubercular activity (MIC 62.5 μg/mL). Further chemical investigation of Arctium lappa led to the isolation of n-nonacosane (1), taraxasterol acetate (2), taraxasterol (3), a (1:1) mixture of β sitosterol/stigmasterol (4), isololiolide (5), melitensin (6), trans-caffeic acid (7), kaempferol (8), quercetin (9), kaempferol-3-O-glucoside (10). Compounds isolated from Tussilago farfara were identified as a (1:1) mixture of β sitosterol/stigmasterol (4), trans-caffeic acid (7), kaempferol (8), quercetin (9), kaempferol-3-O-glucoside (10), loliolide (11), a (4:1) mixture of p-coumaric acid/4-hydroxybenzoic acid (12), p-coumaric acid (13). All compounds were identified following analyses of their physicochemical and spectroscopic data (MS, ¹H and ¹³C-NMR) and by comparison with published data. This is the first report of the isolation of n-nonacosane (1), isololiolide (5), melitensin (6) and kaempferol-3-O-glucoside (10) from Arctium lappa, and of loliolide (11) from Tussilago farfara. Amongst the isolated compounds, the best activity was observed for p-coumaric acid (13) (MIC 31.3 μg/mL or 190.9 μM) alone and in mixture with 4-hydroxybenzoic acid (12) (MIC 62.5 μg/mL).

Conclusions

The above results provide for the first time some scientific evidence to support, to some extent, the ethno-medicinal use of Arctium lappa and Tussilago farfara as traditional antitubercular remedies.     

Identification of trans-tiliroside as active principle with anti-hyperglycemic, anti-hyperlipidemic and antioxidant effects from Potentilla chinesis

Aim of the study

The present study was carried out to isolate and identify trans-tiliroside as principal compound with anti-hyperglycemic, anti-hyperlipidemic and antioxidant effects from Potentilla chinesis.

Materials and methods

A bioactive compound, trans-tiliroside was isolated from the ethanol extract of Potentilla chinesis and its administration dose was optimized and patented. The normal, alloxan-induced diabetic mice and streptozotocin-induced diabetic rats were used to evaluate the anti-hyperglycemic, anti-hyperlipidemic and antioxidant effects of trans-tiliroside from Potentilla chinesis. Biochemical parameters were assayed in blood samples of different groups of alloxan-induced diabetic mice and streptozotocin-induced diabetic rats.

Results

The level of fasting serum glucose levels, triglycerides (TG) and total cholesterol (TC) in alloxan-induced diabetic mice were significantly decrease after daily oral administration of trans-tiliroside in normal and diabetic mice at doses of 0.4, 0.8 and 1.6 mg/kg/day, for 15 days. Blood glucose level was significant decrease in STZ induced diabetic rats by trans-tiliroside (1.2 and 0.3 mg/kg body weight for 10 weeks). The content of TC, low density lipoprotein (LDL-C) and TG levels were decreased and high density lipoprotein (HDL-C) content was increased, so lipid metabolism was improved. Moreover, trans-tiliroside revealed antioxidant activity as shown by increased activities of superoxide dismutase (SOD), decreased level of malondialdehyde (MDA) in diabetic rats. Histological morphology examination showed that the trans-tiliroside restored the damage of pancreas tissues in rats with diabetes mellitus.

Conclusion

Trans-tiliroside, a constituent from Potentilla chinesis, revealed significant anti-hyperglycemic, anti-hyperlipidemic and antioxidant activities.     

Cinchonain Ib isolated from Eriobotrya japonica induces insulin secretion in vitro and in vivo

Aims of the study

Eriobotrya japonica leaves had been used traditionally for the treatment of diabetes mellitus by immersing the dried leaves in a hot water drink. Few studies have shown the hypoglycemic effect of Eriobotrya japonica using crude alcoholic extract and isolated methanolic compounds. These studies proposed that the mechanism of action could be by stimulating the β-islets of Langerhans to secrete insulin, however with no scientific evidence.

Methods

Eriobotrya japonica water extract (EJWE) and the compounds derived from it: cinchonain Ib, procyanidin B-2, chlorogenic acid and epicatechin, were tested for their effects on insulin secretion from INS-1 cells and following oral administration in rats.

Results

The present study showed that EJWE increased significantly (p < 0.05) insulin secretion from INS-1 cells in dose-dependent manner. Oral administration of EJWE at 230 mg/kg to rats, however, decreased plasma insulin level for as long as 240 min post-administration and caused a transient drop of blood glucose at 15 and 30 min post-administration. On the other hand, cinchonain Ib enhanced significantly (p < 0.05) insulin secretion from INS-1 cells, whereas epicatechin inhibited significantly (p < 0.05) insulin secretion from INS-1 cells. In addition, cinchonain Ib enhanced significantly (150%: p < 0.05) plasma insulin level in rats for as long as 240 min after 108 mg/kg oral administration but did not induce any change in blood glucose level.

Conclusion

These data indicate that cinchonain Ib has an insulinotropic effect and suggest the possible use of cinchonain Ib for managing type 2 diabetes.     

Artemisia copa aqueous extract as vasorelaxant and hypotensive agent

Ethnopharmacological relevance

Artemisia copa Phil. (Asteraceae) is a medicinal plant commonly used in traditional medicine in Argentina.

Aim of the study

The vasorelaxant and hypotensive activities of the aqueous extract of Artemisia copa have been investigated.

Materials and methods

The in vitro effect of the extract and isolated compounds from Artemisia copa was investigated using isolated rat aortic rings. The acute effect caused by the intravenous (i.v.) infusion (0.1–300 mg/kg) on blood pressure and heart rate was evaluated in spontaneous hypertensive rats. In addition, a phytochemical analysis of the extract was performed by HPLC.

Results

Artemisia copa had a relaxant effect in endothelium-intact aortic rings that had been pre-contracted with 10⁻⁷ M phenylephrine (E_max=96.7±1.3%, EC₅₀=1.1 mg/ml), 10⁻⁵ M 5-hydroxytriptamine (E_max=96.7±3.5%, EC₅₀=1.5 mg/ml) and 80 mM KCl (E_max=97.9± 4.4%, EC₅₀=1.6 mg/ml). In denuded aortic rings contracted by phenylephrine, a similar pattern was observed (E_max=92.7±6.5%, EC₅₀=1.8 mg/ml). l-NAME, indomethacin, tetraethylammonium and glibenclamide were not able to block the relaxation induced by the extract. Nevertheless, the pre-treatment with Artemisia copa attenuated the CaCl₂-induced contraction in a concentration-dependent manner (E_max: 86% of inhibition for 3 mg/ml and 52% de-inhibition for 1 mg/ml). This pre-treatment also induced a significant attenuation of the norepinephrine-induced contraction in a concentration-dependent manner (E_max: 72.7% of inhibition for 3 mg/ml and 27% de inhibition for 1 mg/ml) in a Ca²⁺ free medium. Upon analyzing the composition of the extract, the presence of p-coumaric acid, isovitexin, luteolin and chrysoeriol were found. Luteolin (CE₅₀: 1.5 μg/ml), chrysoeriol (CE₅₀: 13.2 μg/ml) and p-coumaric acid (CE₅₀: 95.2 μg/ml), isolated from the aqueous extract, caused dilatation of thoracic aortic rings pre-contracted with phenylephrine. Artemisia copa administered i.v. also induced a decrease in the mean arterial pressure but did not affect the heart rate in hypertensive rats.

Conclusions

The aqueous extract of Artemisia copa proved to have vasorelaxing and hypotensive effects through the inhibition of Ca²⁺ influx via membranous calcium channels and intracellular stores. The presence of luteolin, chrysoeriol and p-coumaric acid found in this plant could be involved in this effect.